Research Report 2011: Environmental Biotechnology Research Group

ENVIRONMENTAL BIOTECHNOLOGY RESEARCH GROUP
RESEARCH REPORT 2011

EB GROUP
Environmental Biotechnology Research Group (EB Group), Faculty of Biotechnology and
Biomolecular Sciences, Universiti Putra Malaysia was officially started in 2005. Currently,
EB Group consists of four subgroups; Biocompost, Biomaterial, Bioproduct, Bioenergy.
Biocompost and Biomaterial are led by Professor Dr Mohd Ali Hassan, whereby the other
two groups are led by Professor Dr Suraini Abd Aziz. There are 6 principal researchers in
the group and 33 students including Masters and PhD. Our aim is to be a high performance
research group conducting research on the utilization of oil palm biomass and other
renewable raw materials in Malaysia for valuable green products. Most of our research are
On the cover
conducted in close collaboration with other academic institutions and industries locally
and internationally, such as FELDA, Kyushu Institute of Technology (Kyutech, Japan), and 1st Bioplastic Pilot
National Institute of Advanced Industrial Science and Technology (AIST, Japan).  69 Plant in Malaysia
Message from the

01 EB Group Leader research group
group Big Picture

46
02 04 06
CONTENTS
Bioenergy
EB research Group Serdang Biomass Town Organisation Chart
Bioproduct
group member
08 14 15 16 17 66
principal Associate Group 2011 group
Researcher Researcher Staff Alumni Attachment Biomaterial
group achievement
18 42 43 44 82
publication Patent Consultancy Award
Biofertilizer
90 2011 in Pictures
92 group website
▶ Bioplastic Pilot Plant Holding Tank

Message from the
EB Group Leader
AlhamduLillah, praise to ALLAH for His generous favours and blessings for us. I am very happy that over
the years, our Environmental Biotechnology Research Group (EB) has gone from strength to strength.
I am glad to share with you our research report for 2011. We now have four sub-groups comprising
of bioenergy, bioproduct, biomaterials and biofertiliser. We continue to collaborate with Kyushu Insti-
tute of Technology, FELDA Palm Industries Sdn. Bhd., Advanced Institute of Science and Technology
(AIST) Japan, Malaysian Technology Development Corporation (MTDC), Yayasan Pelajaran Johor and
Ajinomoto Corporation Japan. In addition, we have extended our cooperation with Ministry of Housing
and Local Government – National Solid Waste Management Division, Subang Jaya Municipal Council
(MPSJ), Malaysian Agricultural Research and Development Institute (MARDI), Ministry of Agricul-
ture, Forestry and Fisheries (MAFF) Japan, and recently with CES Company, Incheon Korea. We have 6
academic staff, 14 PhD and 17 MS students at the Faculty of Biotechnology and Biomolecular Sciences,
Institute of Bioscience and Faculty of Engineering UPM. In addition we also have 5 PhD students on the
split program with Kyushu Institute of Technology. In total, we have RM 2.76 million in R&D grants. In
terms of output, we successfully published 13 research papers in 2011, with a total of 11.353 Impact Fac-
tors, with another 9 papers which are currently in press. We also filed 4 patents and won 2 gold medals at
22nd International Invention, Innovation and Technology Exhibition (ITEX 2011).
I appreciate the hard work from all EB members in maintaining our high-performance culture.
May ALLAH give us the strength to continue the good work and contribute to the university, the
ummah and the nation.
God bless. Wassalam.
Professor Dr Mohd Ali Hassan
▶ Specific primer designed before DNA walking ▶ Enzymatic analysis using colorimetric procedures ▶ Lignin-degrading bacteria
RESEARCH REPORT 2011 > EB GROUP 01

EB GROUP BIG PICTURE
proce ss tre ate d
f re s h wate r wate r
f ru i t
w a t er
bun ch
h ydrogen
(ffb )
palm oil producti on
mill
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M I LL total
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• oil palm
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c r u d e p al m
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o il (c po )
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t r a dit io n a l t e c h no l o g y
02 RESEARCH REPORT 2011 > EB GROUP

sludge biocom po st
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SERDANG BIOMASS
BIOMASS TOWN BOUNDARIES COLLECTION FEEDSTOCK
GREEN WASTE
UPM - MARDI - MPSJ ( SERI SERDANG TOWN )
MARDI / UPM TPU FARM SCHEDULED GW COLLECTION

[ GW ]
LIVESTOCK MANURE
UPM TPU HUSBANDRY SCHEDULED LM COLLECTION
[ LM ]
administrative area
SCHEDULED & VOLUNTARY USED COOKING OIL

UCO COLLECTION [ UCO ]
landed property
residential college food waste

SCHEDULED COLLECTION
OF food waste [ FW ]
eateries property
recyclable mat
er
ials
UPM : universiti Putra Malaysia
 TO MARKET MARDI : Malaysian Agricultural Research

and Development Institute
MPSJ : Majlis Perbandaran Subang Jaya

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PARASITIC USE - BIOREFINERY PLANT
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SOLID & LIQUID DIGESTATE

PARASITIC USE - MACHINERY ( BIODIESEL )
CTRICITY GENERATED FROM

BIODIESEL
PLANT
BIODIESEL
BIOGAS
BIOGAS)
ANAEROBIC
MANUAL & MECHANICAL SEPARATION DIGESTER
(ad) PLANT
DIGESTATE
NON-RECOVERABLE MATERIAL / Contraries

LANDFILL

Yee Noor Azman
Lian Ngit Mohd Johar
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EB GROUP KEY // GROUP LEADER
06 RESEARCH REPORT 2011 > EB GROUP PRINCIPAL RESEARCHER

Mohd
Azwan
Jenol
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Azreen  
pa
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Zulkhairi
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phd PH.D. STUDENT ra RESEARCH ASSISTANT
msc MASTER STUDENT

EB GROUP RESEARCHER
Professor Dr Mohd Ali Hassan
recent publications
Norjan Yusof, Mohd Ali Hassan, Phang Lai Yee, Meisam Tabatabaei, Mohd Ridzuan Othman,
Minato Wakisaka, Yoshihito Shirai. 2011. Nitrification of high-strength ammonium landfill
leachate with microbial community analysis using fluorescence in situ hybridization (FISH).
Waste Management and Research, Vol. 29 (6), 602-611 pp.
Tabassum Mumtaz, Suraini Abd-Aziz, Nor’Aini Abdul Rahman, Phang Lai Yee, Helmi Wasoh,
Yoshihito Shirai, Mohd Ali Hassan. 2011. Visualization of core-shell PHBV granules of wild
type Comamonas sp. EB172 in vivo under transmission electron microscope. International
Journal of Polymer Analysis and Characterization, Vol. 16 (4), 228-238 pp.
specialisation Isnazunita Ismail, Mohd Ali Hassan, Nor’Aini Abdul Rahman, Chen Sau Soon. 2011. Effect
of retention time on biohydrogen production by microbial consortia immobilised in
Bioprocess Engineering & polydimethylsiloxane. African Journal of Biotechnology, Vol. 10 (4), 601-609 pp.
Environmental Biotechnology Meisam Tabatabaei, Raha Abdul Rahim, Norhani Abdullah, André-Denis G. Wright,
Yoshihito Shirai, Kenji Sakai, Alawi Sulaiman and Mohd Ali Hassan. 2010. Importance of the
methanogenicarchaea populations in anaerobic wastewater treatments. Process Biochemistry,
current research interest Vol. 45 (8), 1214-1225 pp.
Treatment and utilization of biomass, Yung-Hun Yang, Christopher J. Brigham, Charles F. Budde, Paolo Boccazzi, Laura B. Willis,
Mohd Ali Hassan, Zainal Abidin Mohd Yusof, ChoKyun Rha and Anthony J. Sinskey. 2010.
wastes and effluents for the production Optimization of growth media components for polyhydroxyalkanoate (PHA) production from
of bioproducts, bioremediation and organic acids by Ralstonia eutropha. Applied Microbiology and Biotechnology, Vol. 87 (6),
reduction of greenhouse gasses 2037-2045 pp.
Mohd Rafein Zakaria, Hidayah Ariffin, Noor Azman Mohd Johar, Suraini Abd-Aziz, Haruo
h index: 14 Nishida, Yoshihito Shirai and Mohd Ali Hassan. 2010. Biosynthesis and characterization of
poly(3-hydroxybutyrate-co-3- hydroxyvalerate) copolymer from wild-type Comamonas sp.
EB172. Polymer Degradation and Stability, Vol. 95 (8), 1382-1386 pp.
academic qualification
Norjan Yusof, Mohd Ali Hassan, Phang Lai Yee, Meisam Tabatabaei, Mohd Ridzuan Othman,
PhD (Environmental Biotechnology), Masatsugu Mori, Minato Wakisaka, Kenji Sakai and Yoshihito Shirai. 2010. Nitrification of
ammonium-rich sanitary landfill leachate. Waste Management, Vol. 30 (1), 100-109 pp.
University of Okayama, Japan (1997)
M. Phil. (Chemical Engineering),
University of Birmingham, U.K. (1990)
M.Sc. (Food Engineering), 2011 patents
University of Leeds, U.K. (1982)
B.Sc. (Honours) (Chemical Engineering), Mohd Ali Hassan, Hidayah Ariffin, Mior Ahmad Khushairi Mohd Zahari, Mohd Rafein
Zakaria, Jailani Salihon, Mohd Noriznan Mokhtar, Yoshito Shirai. 2011. Renewable Sugars
University of Leeds, U.K. (1980) From Oil Palm Waste. PI2011004440
Mohd Ali Hassan, Azhari Samsu Baharuddin, Alawi Sulaiman, Minato Wakisaka, Haruo
Nishida,Yoshihito Shirai, Mohd Zulkhairi Mohd Yusof, Ezyana, Noriznan Mokhtar and Lim
contacts Siong Hock. 2011. A Method for Treating Oil Palm Biomass. Patent pending: PI 2011000731
+603-8946 7590 Mohd Ali Hassan, Azhari Samsu Baharuddin, Alawi Sulaiman, Ahmad Amiruddin Mohd Ali
and Yoshihito Shirai. 2011. System for Evaporating Final Discharge Wastewater generated in the
alihas@biotech.upm.edu.my Palm Oil Mill. PI2011005385

EB GROUP RESEARCHER
Professor Dr Suraini Abd Aziz
recent publications
Norhayati Ramli, Suraini Abd-Aziz, Mohd Ali Hassan, Noorjahan Banu Alitheen, Kamarulzaman
Kamaruddin and Zoolhilmi Ibrahim. (2011). Molecular cloning and extracellular expression
of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM. African Journal of
Microbiology Research. 5(21): 3475-3482.
Ahmad Muhaimin Roslan, Phang Lai Yee, Umi Kalsom Md. Shah, Suraini Abd-Aziz and Mohd
Ali Hassan. (2011). Production of bioethanol from rice straw using cellulase by local Aspergillus
sp . International Journal of Agricultural Research. 6(2): 188 – 193.
Ezyana Kamal Bahrin, Piong Yeau Seng and Suraini Abd-Aziz. (2011). Effect of oil palm empty
specialisation fruit bunch (OPEFB) particle size on cellulase production by Botryosphaeria sp. in solid state
fermentation. Australian Journal of Basic and Applied Sciences. 5(3): 276-280.
Biochemical Engineering /
Tabassum Mumtaz, Suraini Abd-Aziz, Nor’Aini Abdul Rahman, Phang Lai Yee, Helmi Wasoh,
Enzyme Technology Yoshihito Shirai and Mohd Ali Hassan. (2011). Visualization of core/shell PHBV granules of
wild type Comamonas sp. EB 172 in vivo under transmission electron microscope. International
Journal of Polymer Analysis and Characterization. 16 (4): 228-238.
current research interest
Norhayati Ramli, Suraini Abd-Aziz, Mohd Ali Hassan, Noorjahan Banu Alitheen and
Utilization of lignocellulosic Kamarulzaman Kamaruddin. (2010). Potential Cyclodextrin Glycosyltransferase Producer
from Locally Isolated Bacteria. African Journal of Biotechnology. 9(43): 7317-7321.
biomass for bioenergy and
bioproduct Mohd Rafein Zakaria, Meisam Tabatabaei, Farinazleen Mohd Ghazali, Suraini Abd-Aziz,
Yoshihito Shirai and Mohd Ali Hassan. (2010). Polyhydroxyalkanoate production from
anaerobically treated palm oil mill effluent by new bacterial strain Comamonas sp. EB172.
World Journal of Microbiology and Biotechnology. 26(5): 767 -774.
h index: 10
Dayang Salwani Awg-Adeni, Suraini Abd-Aziz, Kopli Bujang and Mohd Ali Hassan. (2010).
Review: Bioconversion of Sago Residue into Value Added Products. African Journal of
academic qualification Biotechnology . 9(14): 2016 – 2021.
PhD (Biochemical Engineering), Mohd Rafein Zakaria, Hidayah Ariffin, Noor Azman Mohd Johar, Suraini Abd-Aziz, Haruo
Nishida, Yoshihito Shirai and Mohd Ali Hassan. (2010). Biosynthesis and characterization of
University of Wales, Swansea, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer from wild type Comamonas sp.
United Kingdom (1997) EB172. Polymer Degradation and Stability. 95(8): 1382-1386.
MSc. (Biochemical Engineering),
University of Wales, Swansea, Tabassum Mumtaz, Nor Amanlina Yahaya, Suraini Abd-Aziz, Nor’Aini Abdul Rahman, Phang
Lai Yee, Yoshihito Shirai and Mohd Ali Hassan. (2010). Turning Waste to Wealth - Biodegradable
United Kingdom (1993) Plastics Polyhydroxyalkanoates (PHA) from Palm Oil Mill Effluent (POME). Journal of Cleaner
BSc (Hons) (Clinical Biochemistry), Production. 18(14): 1393-1402.
Universiti Kebangsaan Malaysia (1992)
Nurul Kartini Abu Bakar, Suraini Abd-Aziz, Mohd Ali Hassan and Farinazleen Mohd Ghazali.
(2010). Isolation and Selection of Appropriate Cellulolytic Mixed Microbial Cultures for
Cellulases Production from Oil Palm Empty Fruit Bunch. Biotechnology. 9(1): 73 – 78.
contacts
2011 patents
+603-8946 7479 / 7516
Suraini Abd-Aziz, Mohd Ali Hassan, Mohd Sukri Ismail, Razali Sarbini, Nurul Kartini Abu
suraini@biotech.upm.edu.my Bakar, Mohd Faizal Ibrahim, Mohd Nafis Abdul Razak and Phang Lai Yee. 2011. Crude Cellulase
suraini.aziz@gmail.com Cocktail for Lignocellulosic Materials Degradation. PI2011002674.

EB GROUP RESEARCHER
Dr Phang Lai Yee
selected publications
Ahmad Muhaimin Roslan, Phang Lai Yee, Umi Kalsom Md. Shah, Suraini Abdul Aziz and
Mohd Ali Hassan. (2011). Production of Bioethanol from Rice Straw using Cellulase by Local
Aspergillus sp. International Journal of Agricultural Research. 6(2): 188 – 193.
Farah Nadia Omar, Nor Aini Abdul Rahman, Halimatun Saadiah Hafid, Tabassum Mumtaz,
Phang Lai Yee, Mohd Ali Hassan. (2011). Utilization of kitchen waste for the production of
green thermoplastic polyhydroxybutyrate (PHB) by Cupriavidus necator CCGUG 52238.
African Journal of Microbiology. 5 (19):2873-2879.
Yoshihito Shirai and Mohd Ali Hassan. (2008). Pilot-scale recovery of low molecular weight
specialisation organic acids from anaerobically treated palm oil mill effluent (POME) with energy integrated
system. African Journal of Biotechnology, 7, 3900-3905.
Environmental Biotechnology
Ahmad Muhaimin Roslan, Mohd Ali Hassan, Suraini Abd-Aziz and Phang Lai Yee. (2009).
Effect of POME Supplementation on Cellulase Production from Rice Straw by Local Fungi
current research interest Isolates. International Journal of Agricultural Research.
1. Bioconversion of glycerol into bioetha- Mohamad Firwance Basri, Shahrakbah Yacob, Mohd Ali Hassan, Yoshihito Shirai, Mohd Rafein
Zakaria and Phang Lai Yee. (2010). Improved biogas production from palm oil mill effluent by
nol using local isolate a scaled-down anaerobic treatment process. World Journal of Microbiology and Biotechnology,
2. Bioethanol production from glycerol 26, 505-514.
using immobilized cells
3. Upstream and downstream processing Tabassum Mumtaz, Noor Amalina Yahaya, Suraini Abd-Aziz, Nor’Aini Abdul Rahman, Phang
Lai Yee, Yoshihito Shirai and Mohd Ali Hassan. (2010). Turning waste to wealth-biodegradable
for bioplastic production plastics polyhydroxyalkanoates from palm oil mill effluent – a Malaysian perspective. J. Cleaner
4. Upstream process for hydrogen and Prod.
ethanol co-production using biomass
resources Phang Lai Yee,Minato Wakisaka,Yoshihito Shirai andMohd Ali Hassan. Freezing and thawing
technique for the removal of suspended solids and concentration of palm oil mill effluent
(POME). Journal of Chemical Engineering of Japan, 35, 1017-1019 (2002).
h index: 06
Phang Lai Yee, Mohd Ali Hassan, Yoshihito Shirai, Minato Wakisaka and Mohamed Ismail
academic qualification Abdul Karim. Continuous production of organic acids from palm oil mill effluent with sludge
recycle by the freezing-thawing method. Journal of Chemical Engineering of Japan, 36, 707-710
PhD (Environmental Biotechnology), (2003).
Kyushu Institute of Technology, Japan (2004)
Phang Lai Yee, Minato Wakisaka, Yoshihito Shirai andMohd Ali Hassan.Effects of single food
MSc. (Environmental Biotechnology), components on freeze concentration by freezing and thawing technique. Japan Journal of Food
Universiti Putra Malaysia (2001) Engineering, 4, 77-82 (2003).
BSc. (Biotechnology),
Universiti Putra Malaysia (1998) Phang Lai Yee, Minato Wakisaka, Yoshihito Shirai and Mohd Ali Hassan.Effect of sodium
chloride on freeze concentration of food components by freezing and thawing technique.Japan
Journal of Food Engineering, 5, 97-102 (2004)
contacts 2011 patent

+603-8946 7514 Suraini Abd-Aziz, Mohd Ali Hassan, Mohd Sukri Ismail, Razali Sarbini, Nurul Kartini Abu
Bakar, Mohd Faizal Ibrahim, Mohd Nafis Abdul Razak and Phang Lai Yee. Crude Cellulase
phanglaiyee@biotech.upm.edu.my Cocktail for Lignocellulosic Materials Degradation. (Filed in June 2011; PI2011002674).

EB GROUP RESEARCHER
Dr Hidayah Ariffin
Hiroshi Nonaka, Hidayah Ariffin and Masamitsu Funaoka. 2011. Basic Characteristics of
Cellulase Immobilized on Lignophenol. Kobunshi Ronbunshu. Vol. 68 (5), 315-319 pp.
Hidayah Ariffin, Haruo Nishida, Yoshihito Shirai and Mohd Ali Hassan. 2010. Highly selective
transformation of poly[(R)-3-hydroxybutyric acid] into trans-crotonic acid by catalytic thermal
degradation. Polymer Degradation and Stability. Vol. 95 (8), 1375-1381pp.
Hidayah Ariffin, Haruo Nishida, Mohd Ali Hassan and Yoshihito Shirai. 2010. Chemical
recycling of polyhydroxyalkanoates as a method towards sustainable development.
Biotechnology Journal. Vol. 5, 484-492pp.
specialisation
Haruo Nishida, Hidayah Ariffin, Yoshihito Shirai and Mohd Ali Hassan. 2010. Precise
Bioprocess Engineering and Depolymerization of Poly(3-hydroxybutyrate) by Pyrolysis. In: Biopolymers. Ed. Magdy M.
Elnashar. 369-386 pp.
Environmental Biotechnology
Hidayah Ariffin, Haruo Nishida, Yoshihito Shirai and Mohd Ali Hassan. 2009. Anhydride
Production as an Additional Mechanism of Poly (3-hydroxybutyrate) Pyrolysis. Journal of
current research interest Applied Polymer Science, Vol. 111, 323-328 pp.
1. Utilization of oil palm biomass Hidayah Ariffin, Haruo Nishida, Mohd Ali Hassan and Yoshihito Shirai. 2009. Chemical
recycling of polyhydroxyalkanoates as a method towards sustainable development. Journal of
for the production of bioplastics Bioscience and Bioengineering, Vol. 108, s79p.
(PHA) and biobased chemicals
2. Chemical recycling of PHA Hidayah Ariffin, Haruo Nishida, Yoshihito Shirai and Mohd Ali Hassan. 2008. Determination of
3. Direct recovery of crotonic acid Multiple Thermal Degradation Mechanisms of Poly(3-hydroxybutyrate). Polymer Degradation
and Stability, Vol. 93, 1433-1439 pp.
from PHA-producing bacteria by
pyrolysis Hidayah Ariffin, Mohd Ali Hassan, Umi Kalsom Md Shah, Norhafizah Abdullah, Farinazleen
Mohd Ghazali and Yoshihito Shirai. 2008. Production of bacterial endoglucanase from
pretreated oil palm empty fruit bunch by Bacillus pumilus EB3. Journal of Bioscience and
h index: 04 Bioengineering, Vol. 106 (3), 231-236 pp.
academic qualification Hidayah Ariffin, Haruo Nishida, Yoshihito Shirai and Mohd Ali Hassan. 2008. Non-random
degradation behavior of Poly (3-hydroxybutyrate) in pyrolysis. Polymer Preprints, Vol. 49 (2),
PhD (Environmental Engineering), 451p.
Kyushu Institute of Technology, Japan. (2009)
MSc. (Bioprocess Engineering), 2011 patent
Universiti Putra Malaysia (2006)
Bachelor of Engineering (Process and Food), Mohd Ali Hassan, Hidayah Ariffin, Mior Ahmad Khushairi Mohd Zahari, Mohd Rafein
Zakaria, Jailani Salihon, Mohd Noriznan Mokhtar, Yoshihito Shirai. 2011. Renewable Sugars
Universiti Putra Malaysia (2004) From Oil Palm Waste. PI2011004440
contacts 2011 Seminar presented

+603-8946 7515 Asian Congress on Biotechnology 2011 (ACB 2011), Shanghai, China. 11 – 15 May 2011. Title:
Improved physical properties of polyhydroxyalkanoates by different fermentation strategies and
hidayah_a@biotech.upm.edu.my blending with bio-based materials

EB GROUP RESEARCHER
Dr Nor’Aini Abdul Rahman
Nazlina Haiza Mohd Yasin, Nor’Aini Abd Rahman, Hasfalina Che Man, Mohd Zulkhairi Mohd
Yusoff and Mohd Ali Hassan. 2011. Microbial quantification of hydrogen producing bacteria at
different pH from fermented food waste. International Journal of Hydrogen Energy 36: 9571-9580.
Farah Nadia Omar, Nor Aini Abdul Rahman, Halimatun Saadiah Hafid,
Tabassum Mumtaz, Phang Lai Yee, Mohd Ali Hassan. Enhanced production of
polyhydroxyalkanoates by Cupriavidus necator CCUG 52238 utilizing organic acids
from kitchen waste. African Journal of Microbiology Research. 5(19): 2873-2879.
Siti Balkhis Ibrahim, Nor’Aini Abdul Rahman, Rosfarizan Mohamad and Raha Abdul Rahim.
specialisation 2010. Effects of agitation speed, temperature, carbon and nitrogen sources on the growth of
recombinant Lactococcus lactis NZ9000 carrying domain 1 of the aerolysin gene. African J of
Environmental Biotechnology Biotechnology 9(33): 5392-5398.
Halimatun Saadiah Hafid, Nor ‘Aini Abdul Rahman, Farah Nadia Omar, Phang Lai Yee, Suraini
Abd-Aziz, Mohd Ali Hassan. 2010. A comparative study of organic acids production from kitchen
current research interest wastes and simulated waste. Australian Journal of Basic and Applied Sciences 4(4): 639-645.
Fadzillah Ismail, Nor’Aini Abdul Rahman, Suraini Abd-Aziz, Chong Mei Ling, Mohd
Bioconversion of palm oil waste/ biomass Ali Hassan. 2009. Statistical optimization of biohydrogen production using food
and food waste into value-added products waste under thermophilic condition. The Open Renewable Energy Journal 2: 124-131.
e.g. polyhydroxyalkanotes, organic acids,
Zatilfarihiah Rasdi, Nor Aini Abdul Rahman, Suraini Abd-Aziz, Phang Lai Yee, Mei Ling Chong
biofuel and biocompost
and Mohd Ali Hassan. 2009. Optimisation of biohydrogen production from palm oil mill effluent
by natural microflora using response surface methodology. Open Biotechnology Journal. 8: 79 – 86.
h index: 03 Sim Kean Hong, Yoshihito Shirai, Nor ‘Aini Abdul Rahman and Mohd Ali Hassan. 2009. Semi
and Continuous Anaerobic Treatment of Palm Oil Mill Effluent for the Production of Organic
Acids and Polyhydroxyalkanoates. Research Journal of Environmental Sciences. 3(5): 552 – 559.
academic qualification
Nazlina Haiza Mohd Yasin, Nor Aini Abdul Rahman, Fadzillah Ismail, Mohd Zulkhairi Mohd.
PhD (Metabolic Engineering), Yusof and Mohd. Ali Hassan. 2009. Effect of different temperature, initial pH and substrate
composition on biohydrogen production from food waste in batch fermentation. Asian Journal
Kyushu Institute of Technology, Japan (2004) of Biotechnology. 1(2): 42-50.
MSc (Environmental Biotechnology),
Universiti Putra Malaysia (2000) Farah Nadia Omar, Nor’Aini Abdul Rahman, Halimatun Saadiah Hafid., Phang Lai Yee, Mohd
BSc. (Biochemistry and Microbiology), Ali Hassan. 2009. Separation and Recovery of organic acids from fermented kitchen waste by
an integrated process. African Journal of Biotechnology. 8(21): 5807-5315. ISSN 1684-5315
2011 Seminar presented (poster)

contacts Nor Aini Abdul Rahman and Zatilfarihiah Rasdi. Biohydrogen production from palm oil mill
effluent under control and uncontrol pH. The 3rd International conference on fuel cell & hydro-
+603-8946 6669 gen technology. 2011. 22-23 Nov 2011. Kuala Lumpur
nor_aini@biotech.upm.edu.my

EB GROUP RESEARCHER
Dr Azhari Samsu Baharuddin
Azhari Samsu Baharuddin, Nor’ Aini Abdul Rahman, Umi Kalsom Md Shah, Mohd Ali Hassan,
Minato Wakisaka, and Yoshihito Shirai, (2011). Evaluation of pressed shredded empty fruit
bunch (EFB)-palm oil mill effluent (POME) anaerobic sludge based compost using Fourier
transform infrared (FTIR) and nuclear magnetic resonance (NMR) analysis. African Journal of
Biotechnology 10(41): 8082-8089
Mohd Najib Ahmad, Noriznan Mokhtar, Azhari Samsu Baharuddin, Lim Siong Hock,
Suraini Abd-Aziz, Mohd Ali Hassan and Yoshihito Shirai (2011). Microbial diversity and
physicochemical changes in oil palm frond composting with POME Anaerobic Sludge.
BioResources, 6(4): 4762-4780
specialisation
Biocompost Engineering & 2011 conference / seminar presented
Food and Agro waste utilization Azhari Samsu Baharuddin, Mohd Ali Hassan, Kenji Sakai, Haruo Nishida and Yoshihito Shirai
compost production from empty fruit bunch (EFB) and palm oil mill effluent (POME) sludge
and their recycle. International Symposium on Biomass Refinery in Palm Oil Industry 2011,
current research interest Fukuoka, Japan
1. Tropical biomass technology and Azhari Samsu Baharuddin, Mohd Ali Hassan and Yoshihito Shirai. “Zero discharge system (in
utilization palm oil mills)” & “Strategy for creating business opportunities through zero discharge system”.
Conference on Green Tech for SMEs in Palm Oil and Palm Biomass, 28th July 2011, Sandakan,
2. Biofertiliser production from organic, Sabah
municipal and oil palm waste
3. Zero emission system for palm oil
industry 2011 Research attachment
h index: 02 FELDA Palm Industries Sdn Bhd (Industrial attachment): Biomass utilization from oil palm
biomass-biocompost and biogas production
academic qualification 2011 consultancy

PhD (Bioprocess Engineering),
Working Group Member for “Serdang Biomass Town” project for the production of
Kyushu Institute of Technology, Japan (2010) biocompost, biogas, biochar and biodiesel form MSW and agro waste (collaboration among
MSc (Bioprocess Engineering), UPM, MPSJ, MARDI and Ministry of Housing and Local Government)
B.Eng (Chemical Engineering),
Universiti Kebangsaan Malaysia (2002)
2011 patents
Mohd Ali Hassan, Azhari Samsu Baharuddin, Alawi Sulaiman, Minato Wakisaka, Haruo
Nishida,Yoshihito Shirai, Mohd Zulkhairi Mohd Yusof, Ezyana Kamal Bahrin, Noriznan
Mokhtar and Lim Siong Hock (2011). A Method for Treating Oil Palm Biomass. Patent
contacts pending: PI 2011000731
+603-8946 4424 Mohd Ali Hassan, Azhari Samsu Baharuddin, Alawi Sulaiman, Amir Mohd Ali and Yoshihito
Shirai (2011). System for Evaporating Final Discharge Wastewater generated in the Palm Oil
azharis@eng.upm.edu.my Mill. PI 2011005385

ASSOCIATE RESEARCHER
YOSHIHITO SHIRAI, ph.d. Hiroaki Ogawa Haruo NISHIDA, Dr. Eng

Iyehara, Dr. Phar. Sci.
PROFESSOR PROFESSOR PROFESSOR
Division of Environmental Engineering, Division of Environmental Engineering, Division of Polymer Chemistry for
Biochemical Zero-Emission Bioremediation Circulative Utilization of Ecomaterials
with Precise Kinetics & Computer
Eco-Town Collaborative R&D Center for Department of Biological Functions Simulation Technologies,
Environment and Recycling in Graduate, and Engineering, Chemistry for Environments
School of Life Sciences and Systems Graduate School of Life Science and Systems
Engineering, Engineering, Eco-Town Collaborative R&D Center for
Kyushu Institute of Technology, Japan. Kyushu Institute of Technology, Japan Environment and Recycling in Graduate,
School of Life Sciences and Systems
Engineering,
Tel: +81-93-695-6060 Tel: +81-93-695-6059
Kyushu Institute of Technology, Japan.
E-mail: shirai@life .kyutech.ac.jp E-mail: ogawahi@life.kyutech.ac.jp
http://www.lsse.kyutech.ac.jp/~ecotown/ http://www.life.kyutech.ac.jp/~ogawahi/
Tel: +81-93-695-6233
http://www.life.kyutech.ac.jp/~shirai/html/ ogawalab-frameeng.html
E-mail: nishida@lsse.kyutech.ac.jp
index.html
Umi Kalsom Toshinari Helmi Wasoh @

Md Shah, Ph.D. MAEDA, Dr. Eng. Mohamad Isa, Ph.D.
ASSOCIATE PROFESSOR ASSOCIATE PROFESSOR Senior Lecturer
Department of Bioprocess Technology Division of Environmental Engineering, Department of Bioprocess Technology
Environmental Bio-Adaptation
Faculty of Biotechnology Department of Biological Functions Faculty of Biotechnology and Biomolecular

and Biomolecular Sciences, and Engineering, Sciences
Universiti Putra Malaysia Graduate School of Life Science and Systems Universiti Putra Malaysia
Engineering,
Kyushu Institute of Technology, Japan
Tel: +603-89468478 Tel: +81-93-695-6064 Tel: +603-89468096
E-mail: umi@biotech.upm.edu.my E-mail: toshi.maeda@life.kyutech.ac.jp E-mail: helmi@biotech.upm.edu.my
http://www.life.kyutech.ac.jp/~ogawahi/

Dato Wan Md Zin Yoshito ANDO, Dr. Eng. Minato WAKISAKA, Dr. Eng.
Wan Yunus, Ph.D.
PROFESSOR ASSOCIATE PROFESSOR ASSOCIATE PROFESSOR
Chemistry Department Division of Polymer Chemistry Division of Food Engineering,
Biomass-based Polymer Marine Biomass
Centre For Defence Foundation Studies, Eco-Town Collaborative R&D Center for Graduate School of Life Sciences
National Defence University of Malaysia Environment and Recycling in Graduate, and System Engineering
School of Life Sciences and Systems Kyushu Institute of Technology, Japan
Engineering,
Kyushu Institute of Technology, Japan.
Tel: +603-9051 4421 Tel: +81-93-695-6238 Tel: +81-93-695-6066

E-mail: wanmdzin@upnm.edu.my E-mail: yando@life.kyutech.as.jp E-mail: wakisaka@life.kyutech.ac.jp
GROUP STAFF
Rosema Ghazali Prof. Shin WATANABE Dr S. Karuppuchamy
GROUP SECRETARY FELDA BIOMASS POSTDOCTORAL RESEARCHER

Biomass Technology Centre
PROJECT COORDINATOR Current research interest:
Kyushu Institute of Technology Bioplastics and Bionanocomposites
(Representative Office in Malaysia) Biochar
Tel: +603-8947 1182 Tel: +603-2162 3838 Tel: +603-8946 7593

E-mail: rosema_ghazali@yahoo.com E-mail: wa_shin@streamyx.com E-mail: skchamy@gmail.com

20 1 1
n i
Dr Tabassum DR Mohd Rafein
um
Mumtaz Zakaria @ Mamat
al
Highest qualification with EB Group: Highest qualification with EB Group:
Ph.D (Environmental Biotechnology), Ph.D (Environmental Biotechnology),
UPM, Malaysia UPM, Malaysia
Former Supervisor: Former Supervisor:
Professor Dr Mohd Ali Hassan Professor Dr Mohd Ali Hassan
Current Position: Current Position:

Senior Scientific Officer, Researcher,
Institute of Food and Radiation Biology, SIRIM Berhad, Malaysia
Bngladesh Atomic Energy Commission,
Dhaka, Bangladesh
Isnazunita Nazlina Haiza LIM SIONG

bt Ismail Moh Yasin HOCK
Highest qualification with EB Group: Highest qualification with EB Group: Highest qualification with EB Group:
Master Science of Environmental Master Science of Environmental Master Science of Bioprocess
Biotechnology, UPM, Malaysia Biotechnology, UPM, Malaysia Engineering, UPM, Malaysia
Former Supervisor: Former Supervisor: Former Supervisor:
Professor Dr Mohd Ali Hassan Dr. Nor'Aini Abd Rahman Professor Dr Mohd Ali Hassan
Current Position: Current Position: Current Position:

Senior Researcher, Lecturer, Assistant Manager
SIRIM Berhad, Malaysia School of Allied Health Sciences,
SEGi University College
Nurul Kartini Farah Nadia Halimatun

bt Abu Bakar Omar Saadiah Hafid
Highest qualification with EB Group: Highest qualification with EB Group: Highest qualification with EB Group:
Master Science of Environmental Master Science of Environmental Master Science of Environmental
Biotechnology, UPM, Malaysia Biotechnology, UPM, Malaysia Biotechnology, UPM, Malaysia
Former Supervisor: Former Supervisor: Former Supervisor:
Prof. Dr. Suraini Abd Aziz Dr. Nor 'Aini Abdul Rahman Dr. Nor 'Aini Abdul Rahman
Current Position: Current Position: Current Position:

Associate Monitoring and Research Officer, Research Assistant
Government Liaison, Malaysian Palm Oil Board (MPOB)
Malaysian Technology Development
Corporation (MTDC)

EB GROUP
ATTACHMENT
PARTICIPANTS PROGRAM RESEARCH THEME HOST/LOCATION DURATION SPONSOR
Development of novel Assoc. Professor Kyushu Institute

Dr Hidayah Collaborative Dr Yoshito Ando 10 days
bio-based polymeric of Technology
Ariffin Research
composites
Kyushu Institute of Technology, 24 Oct ~ 3 Nov 2011
(Kyutech), Japan
Japan
Preparation and
Program for local
characterization of
graduate students to
biocomposite from
33 days
undertake overseas 02 Jul ~ 4 Aug 2011
Noor Ida research attachment
polypropylene reinforced
Professor Dr
mesocarp fiber Kyushu Institute
Amalina Yoshihito Shirai of Technology
binti Ahamad Kyushu Institute of Technology,
(Kyutech), Japan
Characterization of Japan
Nordin
Collaborative mesocarp fiber treated 60 days
Research with super heated steam 12 Oct ~ 10 Dec 2011
treatment.
• Steam and super

heated steam hydrolysis Professor Dr Kyushu Institute of
of polyhydroxyalkanoates Haruo Nishida Technology
Elmy Nahida Collaborative Eco-Town Collaborative R&D 83 days
for chemical recycling Center for the Environmental &
binti Othman Research
• Characterization of and Recycling, 10 Feb ~ 10 May 2011
Universiti Kuala
Kyushu Institute of Technology,
steam hydrolyzates using Japan
Lumpur
GPC, NMR and FTIR
Biomass carbonization Professor Dr Kyushu Institute

Mohd Ridzuan Research Attachment Yoshihito Shirai & 36 days
from bamboo by using of Technology
Othman / Training
pond Yamamoto Company 28 Jan ~ 04 Mac 2011
(Kyutech), Japan
Shimane, Japan
OUTBOUND
PARTICIPANTS position RESEARCH THEME PARTICIPANTS position RESEARCH THEME

Associate
Dr. Haruo Professor Dr. Yoshihito Chemical synthsis of
Kyushu Institute of Technology, Chemical recycle professor
Nishida Japan Ando Kyushu Institute of Technology, polymer
Japan
Associate Ph.D student

Dr. Toshinari professor Koichi / staff
Hydrogen fermentation Environment economics
Maeda Kyushu Institute of Technology, Nagata Kyushu Institute of Technology,
Japan Japan
Tatsuya Ph.D student Zero discharge in Kotaro Ph.D student Chemical synthsis of
Kyushu Institute of Technology, Kyushu Institute of Technology,
Yoshizaki Japan palm oil industry Watanabe Japan polymer
Master student Takaaki Master student Chemical synthsis of

Zhang bei Kyushu Institute of Technology, Biomass composite Kyushu Institute of Technology,
Japan Maeda Japan polymer
Hiroki Master student Utilization of Yuki Master student Inhibition of

Kyushu Institute of Technology, Kyushu Institute of Technology,
Ariyoashi Japan Escherichia coli Yoshimizu Japan periodontal pathogen
INBOUND

2011 EB GROUP
PUBLICATIONS
19.963
impact factor score
11.359 8.604
Published in press
impact impact
factor factor
16 cited
journals
04
03
impact group
factor patents
journals filled
 Microbial quantification of Microbial diversity and

IF 0.533 IF 0.573 IF 0.879 IF 4.053
IF 0.573 IF 0.814 IF 1.418


Published hydrogen producing bacteria
at different pH from fermented
physicochemical changes in oil
palm frond composting with
food waste. POME Anaerobic Sludge.
 Nitrification of high-strength
IF 0.188 IF 0.533 IF 0.573 IF 1.222
ammonium landfill leachate with
microbial community analysis
PAGE 20 PAGE 21
using fluorescence in situ hybrid-  Elephantopus scaber induces  Visualization of core/shell
ization (FISH). cytotoxicity in MCF-7 human PHBV granules of wild type
PAGE 22 breast cancer cells via p53-in- Comamonas sp. EB 172 in vivo
duced apoptosis. under transmission electron
 Effect of retention time on microscope.
biohydrogen production by
microbial consortia immobilised
PAGE 23 PAGE 24
in polydimethylsiloxane.
 Enhancement of organic acids  Evaluation of pressed shred-
production from model kitchen
PAGE 25 waste via anaerobic digestion.
ded empty fruit bunch (EFB)-
palm oil mill effluent (POME)
anaerobic sludge based compost
 Utilization of kitchen waste using Fourier transform infra-
for the production of green ther-
moplastic polyhydroxybutyrate
PAGE 26 red (FTIR) and nuclear magnetic
resonance (NMR) analysis.
(PHB) by Cupriavidus necator
CCGUG 52238.  Molecular cloning and
PAGE 28 extracellular expression of

cyclodextrin glycosyltransferase
PAGE 27
gene from Bacillus sp. NR5 UPM.
 Basic Characteristics of
Cellulase Immobilized on
PAGE 29
Lignophenol..
 in press
PAGE 30  Oil Palm Empty Fruit Bunch  Recovery and purification
IF 0.610 IF 1.015 IF 1.879
IF 0.893 IF 1.768
as Alternative Substrate for Ace- of intracellular polyhydroxy-
tone-Butanol-Ethanol Produc- alkanoates from recombinant
tion by Clostridium butyricum Cupriavidus necator using water
 Visible light-sensitive MnO2- EB6. and ethanol.
IF 0.603 IF 0.745 IF 1.091
and CeO2-loaded ZrO2/carbon

cluster/Pt nanocomposite
PAGE 31 PAGE 32
materials.  Separation and purification  Efficient Polyhydroxyal-
of polyhydroxyalkanoates from kanoate Recovery from Re-
PAGE 33 newly isolated Comamonas sp.
EB172 by simple digestion with
combinant Cupriavidus necator
by using Low Concentration of
 Nutrient composition, non- sodium hydroxide. NaOH
starch polysaccharides and true
metabolisable energy value of
PAGE 34 PAGE 35
brown rice as poultry feedstuff.  Biological control of Colle-
totrichum sp. the casual agents of
PAGE 36 anthracnose of red chilli fruits by
 Improved cellulase produc-

using an indigenous streptomy-
ces from Malaysia.  cited
tion by Botryosphaeria rhodina
from OPEFB at low level of mois-
PAGE 37
 Production of Bioethanol
cited
ture condition through statistical from Rice Straw using Cellulase

optimization. by Local Aspergillus sp. Inter-
PAGE 38 national Journal of Agricultural
 patent Research.
PAGE 39
 A Method for Treating Oil  System for Evaporating Final  Effect of oil palm empty
cited
patent patent
patent patent
Palm Biomass. Discharge Wastewater generated fruit bunch (OPEFB) particle

in the Palm Oil Mill. size on cellulase production by
Botryosphaeria sp. in solid state
fermentation.
PAGE 42 PAGE 42 PAGE 40
 Renewable Sugars From Oil  Crude Cellulase Cocktail  Bioconversion of Restaurant
cited
Palm Waste. for Lignocellulosic Materials Waste into Polyhydroxybutyrate

Degradation. (PHB) by Recombinant E.coli
through Anaerobic Digestion.
PAGE 42 PAGE 42 PAGE 41

i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 3 6 ( 2 0 1 1 ) 9 5 7 1 e9 5 8 0
Available at www.sciencedirect.com
journal homepage: www.elsevier.com/locate/he
Microbial characterization of hydrogen-producing bacteria

in fermented food waste at different pH values
Nazlina Haiza Mohd Yasin a, Nor’Aini Abd Rahman a,*, Hasfalina Che Man b,
Mohd Zulkhairi Mohd Yusoff a, Mohd Ali Hassan a
a
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang,
Selangor, Malaysia
b
Department of Biological and Agricultural Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 Serdang, Selangor,
Malaysia
article info abstract
Article history: An anaerobic fermentation of food waste was conducted in a 0.5 L bioreactor incubated at
Received 21 January 2011 a thermophilic temperature of 55 � C to evaluate the effects of different controlled pH values
Received in revised form (5.0, 5.5 and 6.0) on biohydrogen production. Effective biohydrogen production was found
5 May 2011 at controlled pH 5.5 and 6.0 corresponding to lower lactic acid production compared to pH
Accepted 8 May 2011 5.0. It was demonstrated that biohydrogen production from food waste was pH-dependent
with hydrogen yields of 79, 76 and 23 mmol H2/L-media/d for pH 5.5, 6.0 and 5.0, respec-
tively. Specific microbial determination for Clostridium sp. and total bacteria quantification
Keywords: were carried out by the fluorescent in-situ hybridization (FISH) technique. The number of
Biohydrogen Clostridium sp. for acclimatized sludge, fermentation broth at pH 5.0, 5.5 and 6.0 were
Food waste 2.9 � 108, 3.6 � 108, 7.8 � 108 and 5.4 � 108 cells/ml, respectively. The quantification analysis
Thermophilic showed that 92% of the total bacteria belonged to Clostridium sp. from clusters I and XI from
pH the sample at controlled pH 5.5. The denaturing gradient gel electrophoresis (DGGE) bands
Fluorescent in situ hybridization of the sample after heat-treatment, acclimatization and during fermentation indicated the
(FISH) presence of Bacteroidetes, Caloromator australicus sp. and Clostridium sp.
Denaturing gradient gel electropho- Copyright ª 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights
resis (DGGE) reserved.
1. Introduction household solid waste [9] as a substrate contributing to waste

reduction [1,3,10]. The average amount of municipal solid
Hydrogen can be considered as an ideal, clean and sustainable waste generated in Malaysia was 0.5e0.8 kg/person/d in the
energy carrier. It contains the largest gravimetric energy rural areas while it was 1.7 kg/person/d in urban areas [11,12].
density compared to any hydrocarbon fuel. It also can be the The municipal solid waste composition of commercial areas
alternative to fossil fuel since the combustion of hydrogen in Kuala Lumpur, Malaysia consists of 41.48% food/organic
impact factor
produces no greenhouse gases emission [1]. Due to the global waste, followed by plastics (20.98%), paper (18.58%), iron and
nAZLINA HAIZA
environmental issues, interest in biohydrogen production has glass (7.74%), wood and yard (6.71%), and others (4.51%) [12].
MOHD YASIN
increased tremendously from renewable biomass such as Malaysia is now focusing on sustainable development and
4.057
kitchen waste [2e5], palm oil mill effluent [1,6e8] and waste management effectiveness [11]. The use of food waste
2011 ALUMNI - MASTER

BIOENERGY
* Corresponding author. BIOPRODUCT
Tel.: þ6 03 89466699; fax: þ6 03 89467593.
RESEARCH GROUP - BIOHYDROGEN
E-mail address: nor_aini@biotech.upm.edu.my (Nor’Aini A. Rahman).
0360-3199/$ e see front matter Copyright ª 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.ijhydene.2011.05.048

impact factor Mohd Najib
Ahmad
1.418 master student

biofertilizer
research group

Research Article
Waste Management & Research
29(6) 602–611
Nitrification of high-strength ammonium ! The Author(s) 2011

Reprints and permissions:
landfill leachate with microbial sagepub.co.uk/journalsPermissions.nav

DOI: 10.1177/0734242X10397581
community analysis using fluorescence

wmr.sagepub.com
in situ hybridization (FISH)
Norjan Yusof 1,2 , Mohd Ali Hassan 3, Phang Lai Yee 3,

Meisam Tabatabaei 4, Mohd Ridzuan Othman 3, Masatsugu Mori 5,
Minato Wakisaka 1, Kenji Sakai 5 and Yoshihito Shirai 1
Abstract
Nitrification of mature sanitary landfill leachate with high-strength of N NH þ4 (1080–2350 mgL 1) was performed in a 10 L
continuous nitrification activated sludge reactor. The nitrification system was acclimatized with synthetic leachate during feed
batch operation to avoid substrate inhibition before being fed with actual mature leachate. Successful nitrification was
achieved with an approximately complete ammonium removal (99%) and 96% of N NH þ4 conversion to N NO 3 . The
maximum volumetric and specific nitrification rates obtained were 2.56 kg N NH þ4 m 3 day 1 and 0.23 g N NH þ4
g 1 volatile suspended solid (VSS) day 1, respectively, at hydraulic retention time (HRT) of 12.7 h and solid retention time
of 50 days. Incomplete nitrification was encountered when operating at a higher nitrogen loading rate of 3.14 kg
N NH þ4 m 3 day 1 . The substrate overloading and nitrifiers competition with heterotrophs were believed to trigger the
incomplete nitrification. Fluorescence in situ hybridization (FISH) results supported the syntrophic association between
the ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria. FISH results also revealed the heterotrophs as the
dominant and disintegration of some AOB cell aggregates into single cells which further supported the incomplete nitrifica-
tion phenomenon.
Keywords
Nitrification, high-strength ammonium wastewater, mature landfill leachate, biological nitrogen removal, FISH
Date received: 6 September 2010; accepted: 19 December 2010
Introduction 1
Department of Biological Functions and Engineering, Graduate
Biological systems, dr Norjan

namely sequencing batch reactors impact factor
School of Life Science and Systems Engineering, Kyushu Institute of
Technology, Kitakyushu, Fukuoka, Japan.
Yusof
(SBRs), are usually used for landfill leachate treatment in 2
Department of Biology, Faculty of Science and Mathematics,
1.222
Malaysia (Yusof et al., 2009). However, a full-scale SBR Universiti Pendidikan Sultan Idris, Tanjong Malim, Perak, Malaysia.
2009 ALUMNI - PhD 3
Department of Bioprocess Technology, Faculty of Biotechnology and
system for treating landfill leachate may occasionally fail to Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM
remove the ammoniumBIOENERGY BIOPRODUCT
below its permissible limit (10 mgL 1 Serdang, Selangor, Malaysia.
for N NH 3 ), thus theRESEARCH
leaked N GROUP - BIOgas 4
NH þ4 can be found in the Microbial Biotechnology and Biosafety Department, Agricultural
Biotechnology Research Institute of Iran (ABRII), Seed and Plant
nearby river (Yusof et al., 2009). A s an alternative, the phy-
Improvement Institute’s Campus, Karaj, Iran.
sico-chemical method of ammonia stripping has been imple- 5
Department of Plant Resources, Faculty of Agriculture, Kyushu
mented for the pretreatment of N NH þ4 prior to the University, Fukuoka, Japan.
biological methods in order to overcome the inhibitory Corresponding author:
effect of the substrate, mainly the free ammonia (FA) and Norjan Yusof, Department of Biology, Faculty of Science and
22
free nitrous acid (FNA)
RESEARCH on the
REPORT 2011 >
nitrification
EB GROUPprocess. Even so,
occasional release of the ammonia (NH 3) as a result of
Mathematics, Universiti Pendidikan Sultan Idris, 35900 Tanjong
Malim, Perak, Malaysia
Email: norjan@fsmt.upsi.edu.my
Journal of Medicinal Plants Research Vol. 5(24), pp. 5741-5749, 30 October, 2011
Available online at http://www.academicjournals.org/JMPR
ISSN 1996-0875 ©2011 Academic Journals
Full Length Research Paper
Elephantopus scaber induces cytotoxicity in MCF-7

human breast cancer cells via p53-induced apoptosis
Wan Yong Ho1, Swee Keong Yeap2, Chai Ling Ho1, Abdul Rahim Raha1,2,
Abdul Aziz Suraini2,3 and Noorjahan Banu Alitheen1,2*
1
Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra
Malaysia, Serdang 43300 Selangor, Malaysia.
2
Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor Malaysia.
3
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia,
Serdang 43300 Selangor, Malaysia.
Accepted 12 September, 2011
Plants have not only been consumed as food but have also been adopted as folk medicine for centuries.
Elephantopus scaber Linn, a herb from the Asteraceae family, has traditionally been taken as decoction
or tea to cure various ailments and diseases throughout the world. Recent studies had also suggested
that this plant posses various bioactivities such as anti-bacterial, anti-inflammatory, hepatoprotective
as well as anti cancer properties. In this study, the cytotoxic effect of an ethanolic extract of E. scaber
on a breast cancer cell line, MCF-7 and the underlying cell death mechanism was examined. E. scaber
showed cytotoxic effect towards MCF-7 cells with an IC50 value of 15 µg/mL. In comparison to the
untreated control, the extract triggered cell death with increased phosphatidylserine externalization,
DNA breaks and significant morphological apoptotic characteristics in the MCF-cells. Furthermore, we
also found that expression of the tumor suppressor p53 protein was up-regulated in response to the
treatment. In conclusion, these results suggested that the ethanolic extract of E. scaber may be a
potential anti cancer agent for human breast cancer cells by the induction of p53-dependent apoptosis.
Key words: Ethanol extract, MCF-7, tumor suppressor protein, DNA fragmentation, phosphatidylserine
externalization.
INTRODUCTION
Breast cancer is one of the leading causes of cancer cases had also witnessed the susceptibility of women
mortality worldwide every year. Despite the availability of who underwent tamoxifen therapy for more than five
treatments in the form of surgery, radiation therapy, years to have relatively higher risks for stroke, cataracts,
chemotherapy, hormonal therapy and biologic therapy for cardiac arrhythmia or atrial fibrillation,
breast cancers (American Cancer Society, 2007; World hypertriglyceridemia, deep-vein thromboembolic events
Health Organization, 2011), most of the therapeutic and even death (Fisher et al., 2005; Veronesi et al.,
means are associated with some drawbacks such as high 2007). There is indeed a need to search for more easily
cost of treatment and adverse side effects after available and much more reliable therapeutic sources
prolonged exposure. For instance, tamoxifen, the oldest into overcoming the problems associated with current
and most-prescribed selective estrogen receptor breast cancer treatments.
modulator (SERM) for treating hormone-receptor-positive Natural products have been used for a long time in folk
breast cancer, had been proven to reduce the risk of medicine to treat a great variety of diseases. In
developing invasive and non-invasive breast cancer ethnopharmacology, plant-derived products are not only
among women (Fisher et al., 2005). However, clinical useful as medications to cure common injuries and
ailments but are also well known for preventing and
treating dreadful diseases such as cancers (Mann, 2000;
impact factor
*Corresponding author. E-mail: noorjahan@biotech.upm. edu. of natural productsProfessor Dr
Yeap et al., 2010). The ethnopharmacological evidence
usage has therefore become an
my. Tel: +603-89467471. important leadSuraini Abd Aziz
for the discovery of anticancer drugs
0.879 group researcher

BIOENERGY BIOPRODUCT
RESEARCH GROUP

Dr Tabassum impact factor
Mumtaz
2011 ALUMNI - PhD
BIOmaterial
RESEARCH GROUP 0.814
African Journal of Biotechnology Vol. 10 (4), pp. 601-609, 24 January, 2011
Available online at http://www.academicjournals.org/AJB
ISSN 1684–5315 © 2011 Academic Journals
Effect of retention time on biohydrogen production by

microbial consortia immobilised in
polydimethylsiloxane
Isnazunita Ismail1,2* Mohd Ali Hassan1 , Nor’Aini Abdul Rahman1 and Chen Sau Soon2
1
43400 Serdang Selangor, Malaysia.
2
Environmental Technology and Research Centre, SIRIM Berhad, 40911 Shah Alam Selangor, Malaysia.
Accepted 14 December, 2010
An investigation on biohydrogen production from palm oil mill effluent (POME) was conducted in a
continuous stirred tank reactor seeded with polydimethyl-siloxane (PDMS) immobilised mixed cultures
at adjusted retention time. The hydrogen-producing bacteria obtained from an anaerobic digester used
for treating POME were acclimatised and immobilised in PDMS. The immobilised cultures were
assessed for their effectiveness in generating hydrogen in a continuous system. In this study, the
PDMS cultures were fed with raw POME at hydraulic retention times of 6, 4 and 2 days and operated at
controlled pH and temperature of 5.5 and 55°C, respectively. At hydraulic retention time (HRT) 2 days,
the average hydrogen production rate per unit volume of POME was 2.1 NL/L/d. Hydrogen constituted
up to 43% of the total gas produced and methane was not detected throughout the 150 days of
continuous operation. The soluble carbohydrate degradation efficiency was highest at 81.2% during
HRT 4 days and the concentration of soluble metabolites produced, followed the order of acetic >
butyric > ethanol > propionic acid. The microbial diversity of the immobilised consortia determined by
denaturing gradient gel electrophoresis (DGGE) changed at different HRTs; with increasing dominant
species phylogenetically related to Clostridaceae.
Key words: Hydrogen fermentation, palm oil mill effluent, polydimethylsiloxane (PDMS), immobilisation.
INTRODUCTION
Dark fermentation of renewable sources have received broad attention as the process is regarded as green
technology with potential in reducing greenhouse gases
(GHG) impact and relatively low energy intensive com-
pared to the typical hydrogen generation via steam
*Corresponding author. E-mail: isnazunita_ismail@sirim.my. methane reforming (SMR). The GHG emission of hydrogen
Tel: +86-(03)-5544 6559. production via the SMR process is estimated at 9.7 to
13.7 kg CO2/kg H2 produced (Spath and Mann, 2000;
Abbreviations: GHG, Greenhouse gases; SMR, steam Muradov and Vezirolu, 2008) which is one magnitude
methane reforming; POME, palm oil mill effluent; COD, order than other known industrial polluters such as in
chemical oxygen demand; EVA, ethylene vinyl acetate fertiliser, steel and cement production. It is envisaged that
copolymer; PMMA, polymethyl methaacrylate; PVA, polyvinyl the biomass-based hydrogen production by anaerobic
alcohol; PDMS, polydimethylsiloxane; PCR-DGGE, polymerase heterotrophic fermentation is commercially viable by
chain reaction coupled with denaturing gradient gel
electrophoresis; VSS, volatile suspended solid; HRT, hydraulic
integrating the system with current wastewater treatment
retention time; TSS, total suspended solids; VFA, volatile fatty system for concomitant waste reduction and bioenergy
impact factor
acids; TCD, thermal conductivity detector; CPO, crude palm oil; generation (Hawkes et al., 2002).
PEO, polyethylene-octene; ASBR, anaerobic sequencing batch Many research works Isnazunita
on biohydrogen production from
reactor.
Ismail
complex wastewater such as palm oil mill effluent
0.573 2011 alumni - master

RESEARCH GROUP - biohydrogen

African Journal of Biotechnology Vol. 10(65), pp. 14507-14515, 24 October, 2011
ISSN 1684–5315 ©2011 Academic Journals
Enhancement of organic acids production from model

kitchen waste via anaerobic digestion
Halimatun Saadiah Hafid, Nor ‘Aini Abdul Rahman*, Suraini Abd-Aziz and Mohd Ali Hassan
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia
UPM 43400, Serdang, Selangor, Malaysia.
Accepted 27 July, 2011
The aim of this study was to obtain the optimal conditions for organic acids production from anaerobic
digestion of kitchen waste using response surface methodology (RSM). Fermentation was carried out
using 250 ml shake flask which was incubated using an orbital shaker set at 200 rpm. Fermented
kitchen wastes were used as inoculums sources. The individual and interactive effects of pH,
temperature and inoculum size (%) on organic acids production from kitchen waste were investigated.
The highest level of organic acid produced was 77 g/L at optimum pH, temperature, inoculum size of
6.02, 35.37°C and 20% inoculum, respectively. The results indicate that the most significant parameters
affecting the bioconversion of kitchen waste to organic acids were temperature and inoculum size.
Verification experiment of the estimated optimal conditions confirmed that RSM was useful for
optimizing organic acids production from fermented kitchen waste.
Key words: Bioconversion, model kitchen waste, anaerobic fermentation, organic acids, optimization, response
surface methodology.
INTRODUCTION
Kitchen waste and other organic solid waste discharged soluble organic products such as organic acids (Lim et
from households, restaurant, and residue from the food al., 2008). Various organic acids produced from organic
industry make up about 70% of total municipal solid wastes especially kitchen waste has been utilized as
waste (MSW) in Malaysia (Hassan et al., 2001). Due to energy and carbon sources in the production of bio-
its large volume, the disposal of kitchen and organic degradable plastic (Horiuchi et al., 2002). Organic acids
waste will be a big problem. At present, dumping waste in such as lactic acid followed by acetic and propionic acids
landfill is the most common practice. Kitchen waste has were found to be the main products of kitchen waste
high organic content such as soluble sugars, starch, fermentation (Bo et al., 2007). Those organic acids are
proteins, cellulose and etc. (Wang et al., 2005). Since also widely used in pharmaceutical, food and beverage
kitchen waste has high organic compound and moisture industries.
contents, anaerobic process is the most suitable method Apart from that this study also highlights on the
for its treatment in comparison with alternative treatments variation problem of the kitchen waste during anaerobic
such as incineration, landfill and composting (Zinatizadeh digestion process and aims to develop a model of kitchen
et al., 2006). Anaerobic digestion is a complex process waste. The development of model kitchen waste was
that involves hydrolysis, acidogenesis, acetogenesis and important to overcome the variation and fluctuation
methanogenesis (Bo et al., 2007). The advantages of problem of real kitchen waste that later will affect the
anaerobic digestion are volume reduction, waste optimization study of the effect of culture conditions on
impact factor
stabilization and biogas recovery (Wang et al., 2008). organic acids production. In anaerobic digestion, it has
Halimatun
Acidogenesis of kitchen waste produces biogas and been reported that the organic acids production was
influenced by pH, temperature and inoculums size
Saadiah Hafid (Zhang et al., 2008). In acidogenesis process, pH plays a
2011 ALUMNI - master
0.573
significant role because some acid producing bacteria
*Corresponding author. E-mail: nor_aini@ biotech.upm.edu.my. prefer pH ranging from 5.5 to 6.5, to produce acid. The
BIOmaterial
Tel: +603-8946 6699. Fax: +603-8943 0913. disturbance of equilibrium will become toxic to bacteria.
RESEARCH GROUP

African Journal of Biotechnology Vol. 10(41), pp. 8082-8089, 3 August, 2011
Evaluation of pressed shredded empty fruit bunch

(EFB)-palm oil mill effluent (POME) anaerobic sludge
based compost using Fourier transform infrared (FTIR)
and nuclear magnetic resonance (NMR) analysis
Azhari Samsu Baharuddin1, Nor’ Aini Abdul Rahman2*, Umi Kalsom Md Shah2, Mohd Ali
Hassan2, Minato Wakisaka3 and Yoshihito Shirai3
1
Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM Serdang,
Selangor, Malaysia.
2
43400 UPM Serdang, Selangor, Malaysia.
3
Department of Biological Functions and Engineering, Graduate School of Life Science and System Engineering,
Kyushu Institute of Technology, 2-4 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0916, Japan.
Accepted 15 June, 2011
Pressed-shredded empty fruit bunches (EFB) and palm oil mill effluent (POME) anaerobic sludge from a
3
500 m closed anaerobic digester system was utilized for the co-composting treatment. Scanning
electron microscopy (SEM) analysis showed that the shredding-pressing treatment on EFB gave better
results in removing the debris and silica bodies as compared to only shredding treatment. However,
similar characteristics were detected in both physically-treated EFB samples by Fourier transform
-1
infrared (FTIR) analysis, mainly in the regions of 900 to 1740 and 2800 to 3400 cm . After the anaerobic
digestion of fresh raw POME, the protein origin (Amide I) band appeared in the POME anaerobic sludge.
-1
Besides, the band intensities at 2925 and 2855 cm which attributed to the composition of fat and lipid
was decreased. The maturity of the composting material after 40 days of treatment was detected by the
-1
appearance of the nitrate band at 1376 cm and the results corresponded to the final C/N ratio of 12.4.
13
Solid state C CP/MAS nuclear magnetic resonance (NMR) was also used to reveal the characteristic
changes of pressed-shredded EFB-POME anaerobic sludge based compost.
Key words: Empty fruit bunch, palm oil mill effluent, compost.
INTRODUCTION
The co-composting of empty fruit bunches (EFB) and biological process by which organic material undergoes
palm oil mill effluent (POME) is one of the alternative decomposition and transformation processes. Minera-
ways to reduce the abundance of biomass generated at lization and humification of organic matter take place dur-
palm oil mills (Baharuddin et al., 2009). Composting is a ing the composting process. Normally, a compost pile
has two stages: an active composting stage and a curing
period stage. When the temperature rises above 60°C,
many microorganisms begin to die or become dormant,
*Corresponding author. E-mail: nor_aini@biotech.upm.edu.my. and after that the temperature starts to stabilize or
Tel: +603-8946 6699. Fax: +603-8946 0913.
decrease. In the curing stage, the materials degrade at a
Abbreviations: EFB, Empty fruit bunches; POME, palm oil mill much slower rate. The composting process continues
effluent; SEM, scanning electron micrscope; FTIR, Fourier until the remaining nutrients are consumed by the
remaining microorganisms and almost all the carbon is
impact factor
transform infrared; NMR, nuclear magnetic resonance; CPO,
crude palm oil. Dr Azhari
converted to carbon Samsu
dioxide (Kale et al., 2006).

Baharuddin
0.573 group researcher

biofertilizer
research group

African Journal of Microbiology Research Vol. 5(19), pp. 2873-2879, 23 September, 2011
Available online http://www.academicjournals.org/ajmr
Utilization of kitchen waste for the production of green

thermoplastic polyhydroxybutyrate (PHB) by
Cupriavidus necator CCGUG 52238
Farah Nadia Omar, Nor’Aini Abdul Rahman*, Halimatun Saadiah Hafid, Tabassum Mumtaz,
Phang Lai Yee and Mohd Ali Hassan
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia,
43400 Serdang, Selangor, Malaysia.
Accepted 30 May, 2011
Polyhydroxybutyrate (PHB) was produced by Cupriavidus necator CCGUG 52238 using organic acids
from fermented kitchen waste. HPLC and nuclear magnetic resonance (NMR) analyses revealed that the
acid comprised mainly of lactic and acetic acids. In shake flask culture, the lactic acid concentration
above 10 g/L inhibited both cell growth and polyhydroxybutyrate (PHB) production. The PHB
production by the strain was achieved at the highest PHB content of 52.79% in batch fermentation
using the kitchen-waste derived organic acids. The PHB yield and productivity were 0.38 g/g and 0.065
g/L/h, respectively. In fed-batch culture, about 4-fold increase in PHB productivity (0.242 g/L/h) was
achieved by applying intermittent feeding strategy.
Key words: Cupriavidus necator CCGUG 52238, kitchen waste, organic acids, polyhydroxybutyrate (PHB).
INTRODUCTION
The growing interest in the development of biodegradable alone accounted to 3,000 tonnes per day. This will
plastics with properties similar to synthetic thermoplastics continue to increase in coming years and is expected to
for the management of the existing plastic waste has reach 30,000 tons per day in 2020 (GEC). The waste
directed the attention towards bacterial usually consists of 45% food waste, 24% plastic, 7%
polyhydroxyalkanoates (PHA). The production of PHAs paper and 6% iron. In the current practice of waste
from agriculture, food processing waste materials is an management, approximately 95 to 97% of waste
attractive approach for not only effectively decreasing collected is taken to landfill for disposals and less than
PHA production cost but also constructing a process for 5% of the waste is being recycled. However, considering
effective utilization of waste. Considering that PHA con- the landfill shortage and contamination, government
tent and productivity are usually lower for bacteria grown aimed to have 22% of the waste recycled by 2020. While
in crude and inexpensive substrates, the development of composting the kitchen refuse can recycle back the
efficient processes for the successful bioconversion nutrients and energy to soil and reduce almost 50% of
remain a challenge to be pursued (Castilho et al., 2009). the waste; anaerobic digestion can also be applied to
In Peninsular Malaysia, generation of solid waste has convert the food waste into organic acids which are
increased from 17.000 tonnes per day in 2003 to 19,100 preferable substrate for PHA production. Producing PHAs
tonnes in 2005 with an average of 0.8 kilogram per capita
Farah
per day. As of 2008, 23,000 Nadia
tonnes of waste is produced impact factor
from kitchen waste can further reduce the inevitable
usage of conventional plastics when biodegradable PHA
each day in Malaysia,Omar the generation in Kuala Lumpur as packaging material would take place in the market.
0.533
Owing to higher moisture content, kitchen waste is
2011 ALUMNI - master more suitable to undergo anaerobic digestion than
thermo-chemical conversion. Significant research has
*Corresponding author. BIOmaterial
E-mail: nor_aini@biotech.upm.edu.my. been focused on using organic acids from fermented food
RESEARCH
Tel: +603 89466699. Fax: GROUP
603 89430913. wastes as raw materials for PHA production (Hafuka et

African Journal of Microbiology Research Vol. 5(21), pp. 3475-4382, 9 October, 2011
Available online http://www.academicjournals.org/AJMR
Molecular cloning and extracellular expression of

cyclodextrin glycosyltransferase gene from Bacillus sp.
NR5 UPM
Norhayati Ramli1, Suraini Abd-Aziz1*, Mohd Ali Hassan1, Noorjahan Banu Alitheen2,
Kamarulzaman Kamaruddin3 and Zoolhilmi Ibrahim3
1
43400 Serdang, Selangor, Malaysia.
2
Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences,
Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
3
Environment and Bioprocess Technology Centre, SIRIM Berhad, No. 1, Persiaran Dato’ Menteri, Section 2,
40911 Shah Alam, Malaysia.
Accepted 26 August, 2011
The cloning of a polymerase chain reaction (PCR) gene fragment from Bacillus sp. NR5 UPM isolated
from the soil in Malaysia into an Escherichia coli expression vector was successfully carried out.
Analysis of the nucleotide sequences revealed the presence of an open reading frame of 2112 bp which
encoded a protein containing 704 amino acids with a putative molecular weight of 78.6 kDa. The
deduced amino acids sequence showed about 98% homology with the CGTase from Bacillus sp.
KC201. Compared to the wild type, the CGTase that was produced in E. coli cells only required one-
fourth of culture time and neutral pH to produce CGTase. After 12 h of cultivation, the CGTase activity
in the culture medium reached 29.6 U/ml, which was approximately 2.5-fold higher than the CGTase
from the parental strain. The CGTase was produced extracellularly by E. coli (94%) indicating the signal
peptide was functional in E. coli.
Key words: Molecular cloning, nucleotide sequence, cyclodextrin glycosyltransferase, Bacillus sp. NR5 UPM.
INTRODUCTION
Cyclodextrin glycosyltransferase (CGTases, 1,4-α-D- genetic information in order to provide a better CGTase
glucopyranosyltransferase (cyclizing), EC 2.4.1.19) is an production method. The over expression of CGTase
important enzyme that catalyzes the formation of α-CD, genes could enhance the enzyme activity, reduce
β-CD and γ-CD, containing 6, 7 and 8 glucose residues cultivation time and produce less contaminating proteins
linked with α-1,4-glucosidic bonds, respectively. Due to compared to wild type (Charoensakdi et al., 2007). In this
their unique abilities to form inclusion complexes with a study, we have succeeded in isolating the CGTase gene
variety of hydrophobic materials and to entrap volatile from Bacillus sp. NR5 UPM. The isolated CGTase gene
compounds, these CDs have found extensive was cloned into an E. coli expression vector and over
applications in food, pharmaceuticals, agricultural expressed to study the improved properties of the
chemicals, cosmetics, industrial chemicals and others enzyme.
(Hashimoto, 2002).
Recently, many researchers have studied the
molecular cloning of CGTase genes and analysed the MATERIALS AND METHODS
Bacterial strain, plasmids and media
Bacillus sp. NR5 UPM was grown in Horikoshi medium II,
impact factor
*Corresponding author. E-mail: suraini@biotech.upm.edu.my. containing 1% (w/v) soluble starch, 0.5% (w/v) yeast extract, 0.5%
Tel: +6-03-89467516/7479. Fax: +6-03-89467510. Norhayati
(w/v) peptone, 0.1% (w/v) K2HPO4, 0.02% (w/v) MgSO4.7H2 O and
Ramli
0.533 phd student

RESEARCH GROUP - bioproduct

Dr Hidayah impact factor
Ariffin
group researcher
BIOmaterial
RESEARCH GROUP 0.188
Appl Biochem Biotechnol (2012)
Oil Palm Empty Fruit Bunch as Alternative Substrate

for Acetone-Butanol-Ethanol Production by Clostridium
butyricum EB6
Mohamad Faizal Ibrahim • Suraini Abd-Aziz •

Mohamad Nafis Abdul Razak • Phang Lai Yee •
Mohd Ali Hassan
IN PRESS
© Springer Science+Business Media, LLC 2012
Abstract Acetone-butanol-ethanol (ABE) production from renewable resources has

been widely reported. In this study, C. butyricum EB6 was employed for ABE
fermentation using fermentable sugar derived from treated oil palm empty fruit bunch
(OPEFB). A higher amount of ABE (2.61 g/L) was produced in a fermentation using
treated OPEFB as the substrate when compared to a glucose based medium that
produced 0.24 g/L at pH 5.5. ABE production was increased to 3.47 g/L with a yield of
0.24 g/g at pH 6.0. The fermentation using limited nitrogen concentration of 3 g/L
improved the ABE yield by 64%. The study showed that OPEFB has the potential to be
applied for renewable ABE production by C. butyricum EB6.
Keywords Clostridium butyricum - Acetone Butanol Ethanol (ABE) - Oil palm empty
fruit bunch (OPEFB) - Anaerobic fermentation - Biomass
impact factor Mohamad Faizal

Ibrahim
1.879 master student

RESEARCH GROUP - biobutanol
in press
Biotechnol Lett
DOI 10.1007/s10529-011-0783-5
ORIGINAL RESEARCH PAPER
Recovery and purification of intracellular

polyhydroxyalkanoates from recombinant
Cupriavidus necator using water and ethanol
Mitra Mohammadi • Mohd Ali Hassan •
Lai-Yee Phang • Hidayah Ariffin •

Yoshihito Shirai • Yoshito Ando
Received: 22 September 2011 / Accepted: 12 October 2011

Springer Science+Business Media B.V. 2011
Abstract A new halogen-free and environmental- in comparison to the established chloroform extraction
friendly method using water and ethanol is developed method. After optimisation, our results showed that the
as an alternative for the recovery of polyhydroxyalk- halogen-free method produced a PHA with 81% purity
anoates (PHA) from recombinant Cupriavidus necator and 96% recovery yield, in comparison to the chloro-
form extraction system which resulted in a highly pure
PHA with 95% yield. Although the purity of the PHA
using the new method is lower, the molecular weight of
the extracted PHA is not compromised. This new
M. Mohammadi M. A. Hassan (&) L.-Y. Phang method can be further developed as an alternative and
H. Ariffin more environmental-friendly method for industrial
Department of Bioprocess Technology, Faculty of
application.
Biotechnology and Biomolecular Sciences, Universiti
Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
e-mail: alihas@biotech.upm.edu.my Keywords Bioplastic recovery
M. Mohammadi Cupriavidus necator Halogen-free method
e-mail: mitramohammadi@gmail.com Polyhydroxyalkanoates
L.-Y. Phang
e-mail: phanglaiyee@biotech.upm.edu.my
H. Ariffin
e-mail: hidayah_a@biotech.upm.edu.my Introduction
M. A. Hassan
Polyhydroxyalkanoates (PHA) are biodegradable,
Department of Process and Food Engineering,
Faculty of Engineering, Universiti Putra Malaysia, biocompatible, thermoplastic and piezoelectric poly-
43400 UPM Serdang, Selangor, Malaysia mers which are accumulated in various microorgan-
isms as intracellular carbon and energy storage
Y. Shirai Y. Ando
materials during unfavorable growth conditions (Yang
Graduate School of Life Sciences and System
Engineering, Kyushu Institute of Technology, 2-4 et al. 2011). However, PHA has not been successfully
Hibikino, Wakamatsu-ku, Kitakyushu 808-0196, Japan commercialized due to its high costs as compared to
Y. Shirai
Mitra
e-mail: shirai@life.kyutech.ac.jp impact factor
conventional plastics. As an intracellular product, the
recovery of PHA contributes significantly to the
Mohammadi overall economics. Therefore developing an inexpen-
1.768
Y. Ando
sive and less-polluting PHA recovery method to
phd student
e-mail: yando@life.kyutech.ac.jp
BIOmaterial 123
RESEARCH GROUP
in press

Superlattices and Microstructures 51 (2012) 239–246
Contents lists availabl e a t SciVerse ScienceDi rect
Superlattices and Microstructures

journal homepage: www.elsevier.com/locate/superlattice s
Visible light-sensitive MnO 2 - and CeO 2 -loaded ZrO 2 /carbon

cluster/Pt nanocomposite materials
b,*
H. Matsui a , M . Ikegami a, S. Karuppuchamy , M.A. Hassan b, M . Yoshihara a
a
Department of Applied Chemistry, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka,
Osaka 577-8502, Japan
b
Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
a r t i c l e i n f o a b s t r a c t
Article history: Nano-sized ZrO 2 /carbon cluster composite materials (I c’s) were suc-
Received 13 September 2011 cessfully prepare d by the calcination of ZrOCl 2 /starch complex I. I c’s
Accepted 18 November 2011 were found to reduce methylene blue under the irradiation of visible
Available online 30 November 2011
light (k > 460 nm). The materials obtained by calcinin g at 400 and
500 C were selectively loaded with Pt particles to obtain Pt-loaded
Keywords:
ZrO 2 /carbon cluster composite materials denoted as I c400 Pt and
Carbon cluster
Metal oxide
I c500 Pt, respectively. In addition, the resultant materials were modi-
Nanocomposite fied with MnO 2 and CeO 2 particles to achieve MnO 2 - and CeO 2 -
Sensitization loaded ZrO 2 /carbon cluster/Pt composite materials denoted as
Photocatalytic activity I c400 PtMn, I c500 PtMn, I c400 PtCe and I c500 PtCe, respectively. The metal
oxides-loaded ZrO 2 /carbon cluster/Pt composite materials thus syn-
thesized could decompose an aqueous silver nitrate solution by vis-
ible light irradiation to give Ag and O 2 with the [Ag]/[O 2 ] ratios of ca.
4. Visible light-irradiated water splitting examinations with
I c400 PtMn and I c400 PtCe were also investigated and found to yield
H 2 and O2 with the [H 2 ]/[O 2 ] ratios of ca. 2.
2011 Elsevier Ltd. All rights reserved.
1. Introduction
impact factor
Multi-electron transfer by photo-irr adiation has been attracting the attention of chemists in view-
Professor Dr
point of electronic and/or magnetic devices production, artificial photosynthesis, solar cell production,
Mohd
oxidation–red uction, and so on. Various photoche mical effects have Alireported
been Hassan with the use of TiO 2
1.091
and modified inorganic/or ganic compounds, however, only under the irradiation of limited wave-
group researcher
length [1–8] . W e have considered that such a function could be achieved by the combination of
BIOmaterial biofertilizer
RESEARCH GROUP
* Corresponding author. Tel.: +603 8946 7590; fax: +603 8946 7593.
E-mail address: skchamy@ibs.upm.edu.my (S. Karuppuchamy).
0749-6036/$ - see front matter

doi: 10.1016/j.spmi.2011.11.016
in press
2011 Elsevier Ltd. All rights reserved.

3b2 Version Number : 7.51c/W (Jun 11 2001)
File path : p:/Santype/Journals/TandF_Production/Lsst/v47n3/LSST615788/lsst615788.3d
Date and Time : 19/12/11 and 12:07
Separation Science and Technology, 47: 1–8, 2012

Copyright # Taylor & Francis Group, LLC
ISSN: 0149-6395 print=1520-5754 online
DOI: 10.1080/01496395.2011.615788
Separation and Purification of Polyhydroxyalkanoates from

Newly Isolated Comamonas sp. EB172 by Simple Digestion
with Sodium Hydroxide
Mitra Mohammadi,1 Mohd Ali Hassan,1,2 Yoshihito Shirai,3 Hasfalina Che Man,4
5 Hidayah Ariffin,1 Lian-Ngit Yee,1 Tabassum Mumtaz,1 Mei-Ling Chong,1 and
Lai-Yee Phang1
1
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences,
Universiti Putra Malaysia, Selangor, Malaysia
2
Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia,
10 Selangor, Malaysia
3
Graduate School of Life Sciences and System Engineering, Kyushu Institute of Technology,
Kitakyushu, Japan
4
Department of Biological and Agriculture Engineering, Faculty of Engineering, Universiti Putra
Malaysia, Selangor, Malaysia
petrochemical-based plastic waste accumulation. However,

15 A simple, mild, and effective process for the recovery of intra- extensive application of PHA is hindered mainly by their
cellular polyhydroxyalkanoate from a newly isolated gram-negative high production cost compared to conventional plastics 40
wild-type bacteria Comamonas sp. EB172 was developed using (1,2). It has been reported that the upstream and down-
sodium hydroxide. Various parameters such as sodium hydroxide stream processing of bacterial PHA are the key cost factors
concentration, digestion time, and temperature were examined for
20 their effect on polyhydroxyalkanoate recovery. The results showed in the fermentation system (3). Therefore, application of a
that polyhydroxyalkanoate with 88.6% purity and 96.8% recovery suitable strain which can utilize the low-cost carbon
yield were obtained by incubating the dried cells with 0.05 M sodium sources can significantly decrease the PHA production 45
hydroxide at 4� C for 1 h, followed by purification steps using etha- cost. Towards the development of zero discharge strategy
nol and water. Removal of non-polymeric cellular materials from for the palm oil industry, it was indicated that organic acid
25 the Comamonas sp. EB172 was increased under alkaline solution
as a result of enhanced cell wall permeability. In addition, the pres- derived from acidogenic fermentation of palm oil mill efflu-
ence of glycerol in the polymer suspension proved that saponification ent (POME) is a promising and potential cheap substrate
of the lipid layer in the bacterial cell wall occurred due to sodium for PHA production (4). In order to lower down the 50
hydroxide reaction. PHA production cost, the development of a local strain
30
which is able to produce the copolymers from mixed
Keywords Comamonas sp. EB172; downstream processing; organic acid, would be an attractive proposition. As an
NaOH digestion; polyhydroxyalkanoates; purifi- effort, newly isolated Comamonas sp. EB172 was found
cation; recovery
to be a suitable microbe for industrial PHA production 55
because of its capability to accumulate poly(3-hydroxybuy-
INTRODUCTION rate-co-3-hydroxyvalerate) [P(3HB-co-3-HV)] copolymers
35 Nowadays, there has been considerable interest for from mixed organic acids (5). Over the fermentation pro-
usage of biodegradable polymers such as polyhydroxyalk- cess itself, downstream processing also has a significant
anoates (PHAs) to address concerns over the effect on the economics of PHA production. Therefore, it 60
is important to develop a novel, efficient, and less polluted
process for PHA recovery with high purity and recovery
Received 26 AprilMitra
2011; accepted 15 August 2011. impact factor
yield from microorganism cells.
Different methods are available in literature to recover
Mohammadi
Address correspondence to Mohd Ali Hassan, Department of the PHA. However, the selection of a suitable extraction 65
1.015
Bioprocess Technology, Faculty of Biotechnology and Biomole-
method depends on factors such as the cell type, the com-
phdPutra
cular Sciences, Universiti student
Malaysia, 43400 UPM Serdang,
Selangor, Malaysia. Tel.: þ603-89467590; Fax: þ603-89467593. position of the growth medium, the presence of certain
chemical compounds in the environment, the length of
BIOmaterial
E-mail: alihas@biotech.upm.edu.my
RESEARCH GROUP
1
in press

EES-2011-0255-ver9-Mohammadi_1P.3d 12/12/11 3:44pm Page 1
EES-2011-0255-ver9-Mohammadi_1P
Type: research-article
ENVIRONMENTAL ENGINEERING SCIENCE ORIGINAL ARTICLE
Volume 00, Number 00, 2012
ª Mary Ann Liebert, Inc.
DOI: 10.1089/ees.2011.0255
Efficient Polyhydroxyalkanoate Recovery from Recombinant Cupriavidus

necator by Using Low Concentration of NaOH
Mitra Mohammadi,1 Mohd Ali Hassan,1,2,* Lai-Yee Phang,1 Yoshihito Shirai,3 Hasfalina Che Man,4
Hidayah Ariffin,1 Al Ashraf Amirul,5 and Siti Nor Syairah 5
1
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Malaysia.
2
Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, Serdang, Malaysia.
3
Graduate School of Life Sciences and System Engineering, Kyushu Institute of Technology, Kitakyushu, Japan.
4
Department of Biological and Agriculture Engineering, Faculty of Engineering, Universiti Putra Malaysia, Serdang, Malaysia.
5
School of Biological Sciences, Universiti Sains Malaysia, Penang, Malaysia.
Received: June 21, 2011 Accepted in revised form: August 28, 2011
Abstract
An efficient method for recovering polyhydroxyalkanoate (PHA) from bacterial cell was developed by using a
low concentration of sodium hydroxide (NaOH). In this study, the effectiveness of low-concentration NaOH on
PHA recovery from recombinant Cupriavidus necator was investigated by testing several NaOH concentrations,
in relation to digestion time and reaction temperature. Gas chromatography (GC) analysis showed that PHA
with more than 96% purity and recovery yield can be achieved after the recovery process which involved
treatment of lyophilized cells with 0.05 M NaOH at 4�C for 3 h. The GC analysis was supported by transmission
electron microscope images and associated with considerable release of protein after NaOH addition. The
recovery process developed herein was found effective in recovering PHA even from cells with low PHA
content, with only 13% reduction in molecular weight (Mw). Ultimately, the present method could be an
alternative to the PHA recovery by organic solvents, with added values such as being simple, nontoxic and
environmental friendly.
Key words: Polyhydroxyalkanoate; bioplastics; recombinant Cupriavidus necator; NaOH recovery; downstream
processing
Introduction PHA recovery using organic solvents, chemical reagents,

or surfactants has the drawbacks of high cost and serious
A s an effort to prevent the harmful environmental

effect posed by nonbiodegradable, petrochemical-based
plastics, the development of environmental friendly biode-
pollution. These methods are, therefore, difficult to be
commercialized (Xuejun, 2006). Thus, developing a cleaner
recovery system without the use of solvent is essential to
gradable plastics such as polyhydroxyalkanoate (PHA) has eliminate the usage of halogenated solvents such as chlo-
drawn much attention throughout the world (Choi and Lee, roform. It has been found that some of the chemical re-
2004; Jung et al., 2005). However, their production at in- covery treatments such as alkaline pH shock, anionic
dustrial scale is limited by the high cost compared with detergents such as sodium dodecyl sulfate (SDS), and
conventional petroleum-based plastics. Anton et al. (1995) EDTA permit the partial release of intracellular products
proposed some key strategies to reduce the PHA production (Harrison et al., 1991). Strazzullo et al. (2008) reported a
cost; for example, by improving reactor productivity, em- simplified and effective method for direct PHA recovery
ploying cheaper substrates, and also developing economical from humid biomass of Halomonas campaniensis using SDS.
purification and recovery schemes. Therefore, developing a Apart from SDS, NaOH is also used to recover PHA. NaOH
process that allows a simple, efficient, and less polluted re- causes saponification of the lipid layer in the cell wall and
covery of PHA could be an attractive proposition (Chen et al., increases the cell permeability that helps release the non-
1999). polymeric protein material without rupturing the cells
(Harrison et al., 1991; Jacquel et al., 2008). Choi and Lee
(1999) reported the digestion of a non-PHA biomass of re-
combinant E. coli in 0.1 M NaOH at 30�C for 1 h. More than
*Corresponding author: Department of Bioprocess Technology, Fa-
culty of Biotechnology and Biomolecular Sciences, Universiti Putra
90% purity and recovery yield was obtained from the cells
Malaysia, Selangor, Malaysia. Phone: + 603-89467590; Fax: + 603- with 77% polyhydroxybutyrate (PHB) content. A highly
89467593; E-mail: alihas@biotech.upm.edu.my pure PHA can be obtained using a concentrated alkaline
impact factor Mitra

Mohammadi
0.893 phd student

BIOmaterial
RESEARCH GROUP
in press
©TBC Poultry Science Association, Inc. JAPR379
Brown rice as a potential feedstuff for poultry
M. N. Asyifah,* S. Abd-Aziz,* L. Y. Phang,* and M. N. Azlian†
*Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular

Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia;
and †Strategic Livestock Research Center, Malaysian Agricultural Research
and Development Institute (MARDI), GPO12301, 50774 Kuala Lumpur, Malaysia
Primary Audience: Researchers, Nutritionists, Feed Manufacturers
SUMMARY
Rice, especially brown rice, has the potential to replace corn as a feedstuff for poultry. It is an
inexpensive local feed source with high availability and low production and processing costs.
Two local varieties of brown rice, MR239 and MR257, were investigated for use as feedstuffs
in the poultry industry, including their composition and TME values (using the force-feeding
technique). The MR239 and MR257 varieties of brown rice contained nutrients such as CP, fat,
DRAFT
ash, and carbohydrates. The energy content and amino acid profile of MR239 and MR257 are
reported. The nonstarch polysaccharides in MR239 and MR257 consisted of CF, NDF, ADF,
and acid detergent lignin. The β-glucan and arabinoxylan contents in MR239 and MR257 were
determined. Both varieties of brown rice were found to be potential sources of feed for poultry.
Key words: brown rice, nonstarch polysaccharide, nutrient composition, poultry, true metabolizable
energy
TBC J. Appl. Poult. Res. TBC:1–8
doi:10.3382/japr.2011-00379
DESCRIPTION OF PROBLEM monogastric animals. The feedstuffs available

for poultry include corn [AU1: Please verify
Poultry is one of the most advanced indus- change to corn throughout.][1], soybeans
tries in the livestock sector because it involves [2], barley [3], wheat [4], and rice or rice by-
processes such as the production of poultry products [5]. However, the choice of feedstuff is
meat, eggs, and feeds; feed formulation; the dependent on its availability, quality, and overall
feeding process; and the import and export of cost of production.
poultry productsNurul
or the Asyifah
poultry themselves. Toimpact factor
provide complete nutrition for poultry,
This industry is among the
Mustapha most efficient food feed compounds are mostly based on a mixture
0.745
commodity groups supplying the fast-growing of corn and other ingredients. In Malaysia, a
human population, master student
especially through the con- search for local sources of poultry feed has been
sumption of chicken. To produce
BIOENERGY poultry prod-
BIOPRODUCT underway to replace expensive imported feed-
ucts, the feedstuffRESEARCH
must firstGROUP - BIOproduct
be produced as an stuffs, mainly corn and soybeans. The average
energy and protein source for poultry. Natural amount of corn imported for the Malaysian feed
resources from plant materials are the most industry was reported to be 2.879 million tons
in press
significant ingredients in poultry feedstuffs be- per year from 2002 to 2007 [5]. Alternatively,
cause they are excellent sources of protein for
36
1 RESEARCH
Corresponding REPORT
author: 2011 > EB GROUP
suraini@biotech.upm.edu.my
African Journal of Biotechnology Vol. 10(32), pp. 5979-5987, 4 July, 2011
Isolation, characterization and identification of potential

actinobacteria with antifungal activities towards chilli
anthracnose
Jeffrey Lim Seng Heng1, Umi Kalsom Md. Shah2* and Halizah Hamzah3
1
Strategic Resource Research Centre, Malaysian Agriculture Research and Development Institute, Mardi Headquarters,
Persiaran MARDI-UPM, 43400 Serdang, Selangor, Malaysia.
2
Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Accepted 11 April, 2011
Actinobacteria from the genus of Streptomycetes have been regarded as the most potent producers of
bioactive compounds in the world. In this study, a total of 132 isolates of actinobacteria were isolated
from rhizospheres of various plant species planted at MARDI Langkawi Agro Technology Park,
Malaysia. These isolates were screened for the ability to inhibit the growth of pathogenic fungi,
Colletotrichum capsisi and Colletotrichum gloeosporioides isolated from chilli fruit. From these
screening it revealed that 45 isolates of actinobacteria were able to produce antifungal activity towards
C. capsici, while 67 isolates produced antifungal activity towards C. gloeosporioides. Out of these 132
isolates, 2 of the best antifungal-producer were selected and identified as Streptomyces spp. strain PM2
and PM4. Observation using scanning electron microscope (SEM) showed that the spore surface for
both Streptomyces spp. strain PM2 and PM4 were rough and spiky. Physiological characterization of
both strains showed their ability to grow in 1 to 4% of NaCl, growth temperature of 17 to 35°C and pH of
5 to 11. The ability of these Streptomyces spp. to secrete antifungal compounds may have been related
to the availibility of the carbon sources. These findings suggest that Streptomyces spp. strain PM2 and
PM4 are potential candidate for biocontrol against anthracnose disease.
Keywords: Actinobacteria, Colletotrichum capsici, Colletotrichum gloeosporioides, anthracnose, antifungal

activity.
INTRODUCTION
Colletotrichum capsici and Colletotrichum Soil borne actinobacteria had been known to possess the
gloeosporioides have been known to be the casual ability to produce bioactivities such as enzymes,
agents for the anthracnose disease on chilli fruits world pesticides, herbicides and also antibiotics (Parungao et
wide (Manandhar et al., 1995; Sangchote, 1999). C. al., 2007). Currently, fungicides were sprayed to the fruits
gloeosporioides was the most dominant anthracnose to inhibit the disease, but the used of chemical fungicides
causing agent with its ability to infect other crops and would caused the development of resistance by the
fruits such as yam and mango. Anthracnose attack on fungal and also the risk of polluting the environments.
chilli fruits causes the fruits to be blemishes and thus, The used of biological control agents had been
making it unmarketable. The typical symptoms of suggested as an alternative way of controlling plant
anthracnose attack on chilli fruit are characterized by the diseases (Compant et al., 2005). Jeffrey (2008) has
formation of sunken necrotic lession, with concentric suggested that actinobacteria belonging to the genus of
rings of acervuli. Streptomyces can be used as biocontrol agents for
several plant diseases. Study by Johnson (1954) shows
that Streptomycetes spp. gave significant control towards
pythium root rot of corn and sugarcane. While Lee and
impact factor
*Corresponding author. E-mail: umi@biotech.upm.edu.my Tel:
+603- 89468478. Fax: +603-89467510. Assoc. Prof.isolated
Dr Umi
Hwang (2002) showed the production of antifungal
activity by actinomycete from Korean soil
Kalsom Md Shah
0.610 ASSOCIATE RESEACHER

RESEARCH GROUP - BIOPRODUCT
in press
Ezyana Kamal Bahrin1, Mohamad Faizal Ibrahim1, Mohamad Nafis
Abdul Razak1, Suraini Abd-Aziz*1, Umi Kalsom Md Shah1, Noorjahan Alitheen2

and Madihah Md Salleh3
1 Bioprocess Technology Department, Faculty of Biotechnology and Biomolecular
Sciences, Universiti Putra Malaysia, Malaysia.
2 Cell and Molecular Biology Department, Faculty of Biotechnology and Biomolecular
Sciences, Universiti Putra Malaysia, Malaysia.
3 Department of Biotechnology Industrial, Faculty of Biosciences and Bioengineering,
Universiti Teknologi Malaysia, Malaysia.
 The response surface method was applied in this study 26 to improve cellulase production
from oil palm empty fruit bunch (OPEFB) by Botryosphaeria rhodina. An experimental design
based on two-level factorial was employed to screen the significant environmental factors for
cellulase production. Locally isolated fungus, Botryosphaeria rhodina was cultivated on OPEFB
under solid state fermentation (SSF). From the analysis of variance (ANOVA), initial moisture
content, amount of substrate and initial pH of nutrient supplied in the SSF system significantly
influenced cellulase production. Then, the optimization of the variables was done using the
response surface method according to Central Composite Design (CCD). Botryosphaeria rhodina
exhibited its best performance with high predicted value of FPase enzyme production (17.95
U/g) when the initial moisture content was at 24.32 %, initial pH of nutrient was 5.96 and 3.98g
of substrate. The statistical optimization from actual experiment resulted in a significant
increment of FPase production from 3.26 to 17.91 U/g (5.49 fold). High cellulase production at
low moisture content is a very rare condition for fungi cultured in solid state fermentation.
Keywords cellulase, solid state fermentation, optimization, response surface

methodology
Ezyana Kamal impact factor

Bahrin
PhD student
RESEARCH GROUP - BIOETHANOL 0.603
in press

Ahmad Muhaimin
CITED
Roslan
PhD STUDENT
JOURNAL BIOENERGY BIOPRODUCT

RESEARCH GROUP - BIOETHANOL

Australian Journal of Basic and Applied Sciences, 5(3): 276-280, 2011
ISSN 1991-8178
Effect of Oil Palm Empty Fruit Bunch Particle Size on Cellulase Production by
Botryosphaeria sp. Under Solid State Fermentation
Ezyana Kamal Bahrin, Piong Yeau Seng and Suraini Abd-Aziz
Bioprocess Technology Department, Faculty of Biotechnology and Biomolecular Sciences,

Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
Abstract: Locally isolated Botryosphaeria sp. showed the ablity to produce cellulases (FPase, CMCase
and β-glucosidase) from oil palm empty fruit bunch (OPEFB) as substrate. Different particle sizes
(0.25-0.3 mm, 0.42-0.6 mm, 0.84-1.0 mm and 5.0-10 mm) of OPEFB were investigated under solid
state fermentation on the cellulase production. The highest production of FPase and β-glucosidase were
obtained from OPEFB particle size of 0.42 – 0.60 mm with 3.261 ± 0.011 U/g and 0.115 ± 0.008
U/g, respectively. It was found that among the four different OPEFB particle sizes studied, particle
size of 0.84 – 1.0 mm gave the highest activity of CMCase (8.134 ± 0.071 U/g). Highest
concentration of reducing sugars produced in this experiment was 4.303 ± 0.095 mg/ml.
Key words: substrate particle size, cellulase, solid state fermentation, Botryosphaeria sp.
INTRODUCTION
Oil palm mills produce a large amount of biomass waste from its daily operation. Generally, 17.08 million
tones per annum of oil palm empty fruit bunches (OPEFB) have been produced continuously in 2005 (MPOB
2006). Fully utilization of OPEFB can be achieved by generating value added products such as activated
carbon, enzymes, citric acid and others. OPEFB is categorized as lignocellulosic feedstock since it is rich in
cellulose contents .Moreover the usage of OPEFB as substrate in cellulases production can reduce the operation
cost since substrate cost became one of the major operational costs, representing 30-40% of total production
cost (Tanaka et al., 2006; Zhang et al., 2007).
Insolubility of OPEFB is one of the limitations in submerged fermentation. Solid state fermentation (SSF)
is more capable in producing certain enzymes and metabolites that usually produced with low yield in
submerged fermentation. The bioconversion of OPEFB into polyoses by using SSF resembles the natural
condition of growth for the majority of fungi. Bacteria, yeast and fungi are able to grow on solid substrate but
filamentous fungi are the best adapted for SSF (Krishna, 2005). The hypha of the fungi has the power to
penetrate into the solid substrate.
There are several factors involved in the selection of a suitable substrate for SSF such as macromolecular
structure, particle size and shape, porosity and particle consistency (Krishna, 2005; Tao et al., 1997). The
substrate must be in a limited size range for an optimal production of cellulase. This process can be facilitated
by chipping, milling and grinding the biomass into a fine powder to increase the surface area/volume ratio of
the cellulose particle. An optimal sized particle lead to better nutrient absorption, gas exchange and heat
transfer thus high enzyme production.
The species ofEzyana Kamal
Botryosphaeria fungus attacks woody host and it is described as an endophyte (Crous,
CITED
2006) and pathogen Bahrin
on plants. The ascomycete fungus Botryosphaeria sp. produces a broad range of
lignocellulolytic enzymes such as laccases (Barbosa et al., 1996), pectinases (Da Cunha et al., 2003) , cellulase
and xylanases (DekkerPhD et
student
al., 2001). These enzymes play an important role in the degradation process of
JOURNAL
lignocellulosic materials through a synergistic action (Lynd et al., 2002; Zhang and Lynd, 2004).
Less report areRESEARCH
available GROUP
on cellulases production by Botryosphaeria sp. using lignocellulose material.
- BIOETHANOL
Dekker et al. (2001) only reported low activity of filter paper cellulase in the media containing veratryl
alcohol. The main objective of this study is to evaluate the potential of locally isolated fungus, Botryosphaeria
sp. to produce cellulases under solid state fermentation by investigating the effect of different substrate particle
size.

Corresponding Author: Suraini Abd Aziz, Department of Bioprocess Technology, Faculty of Biotechnology and
Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
E-mail: suraini@biotech.upm.edu.my
276
4 Mt. J. Environment and Waste Management, In Press
Bioconversion of Restaurant Waste into

Polyhydroxybutyrate (PHB) by Recombinant E.coli
through Anaerobic Digestion
Majd Khalid Eshtaya, Nor Aini Abdul

Rahman, Mohd Ali Hassan
Abstract: Organic acids (lactic, acetic and butyric) are the major fermentation
acids produced on acidogenesis of organic waste such as food waste. In this
study, polyhydroxybutyrate (PHB) was produced from restaurant waste in a two-
step process of microbial acidogenesis and acid polymerization. The effect of
temperature (30°C, 37°C and uncontrolled) and initial pH adjustment at pH 7 in
the anaerobic digestion process were investigated to enhance the production of
organic acids from the waste. Anaerobic digestion of blended restaurant waste
was carried out using a 500 mL Erlenmeyer flask agitated on an orbital shaking
incubator. The highest organic acids level obtained were 39.6 g/L on the fifth day
of fermentation conducted at 30°C and initial pH 7. The acids produced
corresponded to 39.4% of the yield based on the initial COD of substrate. The
main organic acids produced were lactic acid and acetic acid while other acids
were produced in small amount. The fermentation acids were tested for
polyhydroxybutyrate (PHB) production by recombinant Escherichia coli
pnDTM2 and the results were comparable to those of mixed pure acids. Using
organic acids from fermented restaurant waste, recombinant E.coli gave PHB
concentration, PHB content and PHB productivity of 9.2 g/L, 44% w/w and 0.54
gL1h1, respectively in a pH stat fed-batch culture.
Keywords: organic acids, anaerobic digestion, restaurant waste,
polyhydroxybutyrate
Dr Nor’Aini
CITED
Abdul Rahman
GROUP RESEACHER
JOURNAL BIOENERGY BIOPRODUCT

RESEARCH GROUP
in press
A Method for Treating Renewable Sugars From
Oil Palm Biomass Oil Palm Waste
Inventor : Inventor :
Mohd Ali Hassan, Azhari Samsu Baharuddin, Alawi Mohd Ali Hassan, Hidayah Ariffin, Mior Ahmad Khushairi
Sulaiman, Minato Wakisaka, Haruo Nishida, Yoshihito Mohd Zahari, Mohd Rafein Zakaria, Jailani Salihon,
Shirai, Mohd Zulkhairi Mohd Yusof, Ezyana Kamal Bahrin, Mohd Noriznan Mokhtar and Yoshihito Shirai
Noriznan Mokhtar and Lim Siong Hock
Abstract :
The invention provides method for producing sugars from oil palm
Abstract :
fronds wherein the steps comprises of (1) extracting oil palm frond juice
The present invention relates to a method for treating oil palm biomass using
from the oil palm fronds; and (2) treating the oil palm frond juice. The
high pressure steam and/or superheated steam available at the palm oil mill
invention also provides a method of using the said oil palm frond juice for
which is operated at temperature range 100°C-350°C and pressure range
producing polyhydroxyalkanoates such as polymer of hydroxyalkanoic acid,
40-450psig (0.27-3.1MPa) for up to 180 minutes of holding time to enhance
hydroxybutyric acid, hydroxyvaleric acid, and a copolymer thereof, wherein
its degradation characteristics for potential use as a feedstock material for
the copolymers maybe poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV),
production of high value-added bioproducts such as biocompost, biogas,
poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P3HB4HB), polymers
biohydrogen, biosugar, bioethanol and others. The oil palm biomass is such as
and/or copolymers of hydroxyterminated polyhydroxybutyrate (PHB-OH),
empty fruit bunches, frond, mesocarp fibres, palm oil mill effluent (POME)
heteropolymers thereof and any other polymers. In addition to that, biofuels
solid and anaerobic sludge or any combination thereof.
and organic acids such as bioethanol, biobutanol, lactic acid, succinic acid,
and all products that can be produced through chemical and biological
synthesis from these sugars from the OPF juice. The present invention also
IP Status : Pending Patent provides a method for producing polyhydroxyalkanoate (PHA) by incubating
Filed Year : 2011-02-17 at least one strain of PHA-producing microorganisms in a culture medium
Application No. : PI2011000731 comprising of sugars and/or a derivative thereof from the oil palm frond juice.
Country Filing : Malaysia The microorganisms used can be of bacteria, mold or yeast from the group
Applicant : Universiti Putra Malaysia (UPM), Azotobacter, Pseudomonas, Coliform, Alcaligenes, Bacillus, Lactobacillus
FELDA Palm Industries (FPI), and genetically modified form thereof preferably Azotobacter chroococcum,
Kyushu Institute of Technology (KIT) recombinant Escherichia coli, Alcaligenes latus, Pseudomonas oleovorans and
Cupriavidus necator.
IP Status : Pending Patent

Filed Year : 2011-09-20
Application No. : PI2011004440
Country Filing : Malaysia
Applicant : Universiti Putra Malaysia (UPM)
EB GROUP
PATENTS
System for Evaporating Crude Cellulase Cocktail

Final Discharge Wastewater for Lignocellulosic
generated in the Palm Oil Mill Materials Degradation
Inventor : Inventor :
Mohd Ali Hassan, Azhari Samsu Baharuddin, Suraini Abd-Aziz, Mohd Ali Hassan, Mohd Sukri Ismail,
Alawi Sulaiman, Amir Mohd Ali and Yoshihito Shirai Razali Sarbini, Nurul Kartini Abu Bakar, Mohd Faizal Ibrahim,
Mohd Nafis Abdul Razak and Phang Lai Yee
Abstract : Abstract :
A system for evaporating final discharged wastewater generated The invention provides enzymes which are useful for degradation of
in a palm oil mill comprising removal of wastewater grit, lignocellulosic materials, which is obtained from Trichoderma asperellum
equalization and stabilization of steam, evaporation, collection UPM1 and Aspergillus fumigatus UPM2 and wherein the steps includes: (a)
of condensed steam and water vapour and, collection and growing of the Trichoderma asperellum UPM1 and Aspergillus fumigatus
distribution of clean water. UPM2; (b) carrying out fermentation process using the fungi grown from
step (a) with substrates which has been treated to obtain enzymes; and (c)
IP Status : Pending Patent recovering the enzymes from fermented products obtained from step (b). The
Filed Year : 2011-11-08 enzymes recovered include cellulases, xylaneses, pectinases or a combination
Application No. : PI2011005385 of these enzymes which can be further used to hydrolyze lignocellulosic
Country Filing : Malaysia materials for fermentable sugars
Applicant : Universiti Putra Malaysia (UPM)
IP Status : Pending Patent
Filed Year : 2011-06-10
Application No. : PI2011002674
Country Filing : Malaysia
Applicant : Universiti Putra Malaysia (UPM),
AlafPutra Biowealth Sdn. Bhd.

consultancy
Clients /
Research theme Duration
Industrial partner
Biomass Technology
Research Centre,
National Institute of
Utilization of Palm Biomass as Feedstock Advanced Industrial renew until
for Biofuels and Biomaterials
Science and Technology march 2013
(AIST) Japan &
Kyushu Institute of
Technology (KIT) Japan
Effective Utilization of Biomass Through Ajinomoto Co. Ltd, renew until

Biotechnology Japan march 2013
Bioconversion of Palm Oil and Its Products to renew until

SIRIM Bhd
Polyhydroxalkanoates (PHA) march 2012
Improved Biogas Production FELDA Proctor Gamble 2007 ~ 2011

from Glycerine Washwater (FPG) Sdn Bhd completed
Production of Biocompost from Oil Palm Empty YPJ Plantation 2009 ~ 2010
Fruit Bunches (EFB) and Goat Manure Sdn Bhd completed
Feasibility Study on CDM Biomass Projects Tokyo Electric Power 2008

for Renewable Energy Company (TEPCO), Japan completed
Improvement of Wastewater Treatment 2006 ~ 2008

System in Miri Crude Oil Terminal Petronas
completed
Microbial Production of Ethanol Idemitsu Co. Ltd., 2007

from Glycerine Wastewater Japan completed
Production of Activated Carbon and 2005 ~ 2007

NewTech Sdn. Bhd
Pyroligneous Acid from Palm biomass completed
Industrial Grant Scheme (IGS) on

2003 ~ 2005
Bioplastics, Bioacids and Compost Kasa Ganda Sdn. Bhd
from Organic Wastes completed

AWARDS
INTERNATIONAL INVENTION, INNOVATION

& TECHNOLOGY EXHIBITION, ITEX 2011
Kuala Lumpur Convention Centre, Malaysia
Comamonas Putranensis Eco-Friendly

sp. nov., Non-Halogenated
A Novel Bacterium Method for Recovery
Producing Of Intracellular
Polyhydroxyalkanoates Polyhydroxyalkanoates
From Palm Oil Mill (PHA)
Effluent
Prof. Dr. Mohd Ali Hassan, Prof. Dr. Mohd Ali Hassan,
Mr. Mohd Rafein Zakaria, Ms. Mitra Mohammadi,
Mr. Noor Azman Mohd Johar, Dr Phang Lai Yee, Dr. Hidayah
Prof. Dr. Yoshito Shirai, Ariffin, Dr. Amirul Al-Ashraf
Dr. Hidayah Ariffin, Abdullah, Dr. Hasfalina Che Man,
Prof. Dr Suraini Abd. Aziz Prof. Dr. Yoshito Shirai
Gold MEDAL Gold MEDAL
▶ ITEX 2011 - judging process

AWARDS
SHELL INTERVARSITY STUDENT

PAPER PRESENTATION CONTEST 2011
(S-SPEC 2011)
solely sponsored by organized by
4-5 April 2011, University Teknologi Malaysia, Skudai, Johor
Competition: Poster Presentation

Poster Title: Biobutanol as an alternative energy in
developing low carbon economy for Malaysia
Price: Grand Price (First Place)
Name: Mohamad Faizal Bin Ibrahim
Competition: Paper Presentation

Paper Title: Production of bioethanol from rice straw
using Cellulase by Local Aspergillus sp.
Price: First Runner Up
Name: Ahmad Muhaimin Roslan
▶ Faizal was presenting his research findings ▶ Muhaimin's plaque

BIOENERGY
RESEARCH
gineering approaches.
 GROUP LEADER  GROUP INTRODUCTION Currently, there are
T his research group

consists of four
research areas includ-
cultural biomass such
as palm oil mill effluent
(POME), oil palm empty
nine PhD students,
eight Master students
and one research assis-
ing Bioethanol, Biobu- fruit bunch (OPEFB), tant under this group.
tanol, Biohydrogen oil palm decanter cake Our research group
and Bioproduct. Our and sago waste residues emphasizes on develop-
Professor Dr collaborative research into biobased product ing indigenous technol-
Suraini Abd Aziz area focused on the through biotechnology for beneficial bio-
bioconversion of agri- ogy and bioprocess en- energy production using
 RESEARCH AREA
BIOETHANOL BIOBUTANOL
 Bioethanol is an alternative energy source and usually related  Biobutanol is suitable for replacing petrol as fuel in gasoline
to high world market demand. Recently, it has been reported that engines. Besides, it is one of the alternative bioenergy in the next
conversion of biomass into bioethanol involved physical, chemi- future. There are five steps involving utilization of biomass and other
cal and biological methods where the lignocellulosic material will waste materials for biobutanol production including pretreatment
be pretreated by thermo-chemical process, steam or mechani- of biomass, cellulases production, enzymatic hydrolysis, biobutanol
cal grinder before it can be further hydrolyzed into fermentable production and finally is the recovery of biobutanol. In addition,
sugars. The sugars obtained from hydrolysis process can be subse- substrates like oil palm empty fruit bunches (OPEFB), oil palm
quently utilised for bioethanol production using local isolates from decanter cake (OPDC) and sago pith residue (SPR) will have to be
EB culture collection. ✳ hydrolyzed in order to biosugars and later can be further converted
into biobutanol by locally isolated microorganisms from EB
Research Group culture collection. ✳
▶ Bioethanol production using 1liter bioreactor ▶ Biobutanol production from OPDC hydrolysate ▶ Super-Heated Steam Equipment for pretreatement

BIOPRODUCT
GROUP
local resources. On top using several indigenous or enzymatically prior
of that, this research isolates. Bioconversion to actual production/
 PRINCIPAL RESEARCHER
group also involved in of biomass materials fermentation process.
downstream process- into bioenergy and/ Our industrial partners
ing and basic molecular or bioproduct can be include FELDA Palm
biotechnology-based. achieved by one-step or Oil Industry Sdn, Bhd,
Besides, our research two-step fermentation AlafPutra Biowealth
activity has been ex- process that involves Sdn.Bhd and other re-
tended to downstream enzymatic reaction. In search institutes include Dr Phang Dr Nor’Aini
processing and basic some cases, the biomass SIRIM Berhad, MAR- Lai Yee Abdul Rahman
molecular biology study is pretreated chemically DI, KIT and AIST. ✳
BIOHYDROGEN BIOPRODUCT
 It has been known for many years that hydrogen can be  This research group involves in producing different biobased
produced from a variety of renewable resources such as agri- products by utilization of different substrates and microorganisms.
cultural or food industry wastes by using indigenous micro- All the projects apply biological pathways and microbial
organisms. Our research works are focuses on the production fermentation in order to produce beneficial enzymes and flavour
of biohydrogen from sago waste residues as substrates. One of compound as well as hydrolysis process for improvement of
the important factors governing the hydrogen conversion that product quality. Apart from that, this group aims to achieve higher
needs to be taken into account is the engineering process, such enzymatic activity in cyclodextrin glycosyltransferase production
as bioreactor design and operating parameters. Apart from that, via molecular approaches, improving the nutritional value of
co-production of hydrogen and ethanol from biomass becomes poultry feed and production of flavour compound, biovanillin
one of our research focus areas too. We aim to enhance the bio- through microbial fermentation, genetic engineering as well as
mass utilisation efficiency and bioenergy production through recovery of biovanillin. ✳
genetic modification and bioprocess control. ✳
▶ Submerge fermentation for production

▶ Top view of SSF during enzyme extraction ▶ Crude glycerol obtained from palm oil-based biodiesel
of cellulase from OPEFB

Ezyana Zuraidah
 GROUP BIG PICTURE
Cellulase
Ahmad Nur Fairouz

Saleha
Muhaimin Ain Jahaan
Biosugar
Sheril Dayang Rozaimi

Norliana Salwani
Bioethanol Biovanillin
Mohd Nur
Zulkhairi Amelia
Biohydrogen
BIOENERGY BIOPRODUCT Bioproduct
RESEARCH GROUP
Biobutanol
Mohd Mohamad
Azwan Faizal
CGTase
Biosugar Poultry Feed

Mohamad
Nafis
Cellulase
Nurul
Siren Norhayati
Asyifah
▶ Hollow zone produce by bioethanol producing bacteria in screening method for bioethanol

Saleha Shamsudin
b i o ET H A N O L PhD Student, Semester 8
sala_kl@yahoo.com
CELLULOSIC BIOETHANOL FROM STEAM PRETREATED

OIL PALM EMPTY FRUIT BUNCH (OPEFB)
I n the palm oil mill, large amount of by- drolysis. To obtain the most favorable
condition of high pressure steam
RESEARCH OBJECTIVES
about 10% of the total product produced from Steam pretreatment has
dry biomass produced oil palm plantations and been chosen as the most pretreatment that increase OPEFB
by the palm is the oils; palm oil mills. It can be favourable pretreat- digestibility to biosugars production
the other 90% of the used as raw material to ment of OPEFB. This
palm represents a fur- produce bioethanol due pretreatment has to be To obtain high sugars concentration
ther huge source of fiber to it contains of cellu- thought to have rela- from steam pretreated OPEFB by
and cellulosic materials lose and hemicellulose tively moderate energy Acremonium cellulase using
which await a further that can be degraded cost production due Response Surface Methodology
commercial exploita- into fermentable sug- to the steam is already
tion. The availability of ars through enzymatic generated as part of the To obtain high ethanol yield by
this excess energy sourc- hydrolysis. In order to mill operation for elec-
Saccharomyces cerevisie using
es at the mill and its obtain high fermentable tricity and sterilising the
utilization, could helps sugars, the structure of fruit. Besides that, all
Response Surface Methodology
to minimize the cost of OPEFB has to be altered raw materials to initiate
palm oil production in or removed by a suitable the pretreatment (water,
overall. One of alterna- or ideal pretreatment OPEFB, fiber and shells) three main components. the pretreated lignocel-
tive ways in further use in the production of are available in the mill. Heating biomass in the lulosic biomass to more
of these waste materials bioethanol. This crucial This in overall will en- presence of saturated than 80% of sugars
is the utilization in the process is primarily to hance the sustainability steam of 190oC and yielded from enzymatic
area of pretreatment for make the biomass easily of oil palm plantations. 220oC and pressure 1.2 saccharification. In this
bioethanol production. broken down into sugars Furthermore steam to 4.1 MPa normally is study, a variety of op-
Oil palm empty fruit by it opened structure pretreatment is suitable efficient in partially hy- eration temperatures,
bunch (OPEFB) is a through enzymatic hy- to be implementing in drolysed hemicelluloses, pressures and residence
the palm oil mill modified the lignin, times to be applied to
as the OPEFB can increase in accessible the OPEFB have to be
immediately be surface area, decrease tested. In general, these
processed and sac- of the cellulose crystal- parameters however are
charified to the linity and its degrees of different depending on
biosugars for sub- polymerization. There- the pretreatment strate-
sequent bioethanol fore, in this study the gy as well as on the type
production. high pressure steam of and physical of the raw
High pressure 0.8 to 2.3 MPa will be material used to make
steaming is con- studied on the effect of the pretreatment suc-
sidered one of the enzymatic hydrolysis for cessfully, effectively and
most successful op- biosugars production. had a positive im-
tions for fraction- The steam pretreatment pact on the over-
ating wood into its is believed to convert all process. ✳
[SUPERVISOR] Professor Dr Mohd Ali Hassan

Ahmad Muhaimin
Bin Roslan
bioETHANOL
PhD Student, Semester 1
emin85@yahoo.com
ENHANCEMENT OF BIOETHANOL PRODUCTION AND

ADDED VALUE PRODUCTS FROM PALM BIOMASS
I n Malaysia, palm oil

industries play an es-
sential role and deemed
bio-fuels, which include
ethanol, methanol, bio-
diesel and biohydrogen.
and fermentation, SFF).
However saccharifi-
cation required high
ties in ethanol purifica-
tion. And purification
of bioethanol is costly,
the advent of bioengi-
neering to the microor-
ganisms firmly explicit
one of the major con- In the present situation, temperature which will depending on the sub- in to the synthetic media
tributions in agro-in- various attempts have inhibit yeast growth for strate, therefore require or well controlled condi-
dustries, beside of other been made to produce fermentation. suitable technique. tion and possess ex-
commercial crops, like bioethanol as alterna- There are few things to tremely higher yield of
rubbers and cocoas. In tives renewable energy be considered: No. (2) and (3) and (4) product. We are trying
year 2002, approximate- from biomass. Oil palm can be solved through to elucidate the knowl-
ly 11.9 million tones biomass relatively has 1. SFF technique in- metabolic engineering edge to the environmen-
of crude palm oil were potential as a substrate volved mixing enzyme to create novel pathway tal application.
produced in Malaysia. for generation of bioeth- with microorganisms for bioethanol produc- Our laboratory has
at running temperature
In palm oil processing, a anol, hence, the devel- tion. While no. (1) and demonstrated expertise
for the enzyme which
lot of biomass produced opment of an improved (5) require a depth study. in both theoretical anal-
is usually high. A good
as a waste materials and fermentation process thermodynamic balance ysis of metabolic net-
oil palm biomass has for this organic waste is. must be find to stabilize Important of the study works as well as genetic
been identified as one Biological production of microorganisms growth construction of meta-
of the biggest resources ethanol (bioethanol) is and products produc- The study mends to bolically engineered or-
that can be easily find one of the compromis- tion. access appraisal of ganisms. Therefore, we
in a bulk amount Mean- ing technique which is 2. Yeast must survive molecular-based bio- propose to pursue the
while, in the palm oil applicable in the current at saccharification tem- technology technique to following objectives
processing generate situation since the palm perature. circumvent and enhance for the design and con-
3. Utilization of 5-car-
large quantities of solid oil industries manage to fermentation process on struction of an efficient
bon sugar by yeast is
biomass which are rich produce lot of biological very low. bioethanol production microorganism for
in cellulose, hemicellu- substance as a biomass. 4. The microorgan- from harboring sources ethanol produc-
loses and lignin and can isms used are sensitive like oil palm biomass. tion. ✳
be classified as a biomass Background of to the by-products and Putatively,
waste. Biomass is one of the study and to the ethanol itself. This
the materials, which can problem arose will limit the maximum
be used as fuel in en- ethanol production.
ergy industry for future There have been a new Ethanol produced will
demand, and could be- interest to produce be accumulated in the
come largest source of bioethanol simultane- fermentation medium
domestic renewable en- ously as the saccharifica- and only partially sepa-
ergy. Conversion of bio- tion is running (simul- rated. This will
mass can also produce taneous saccharification lead to difficul-
[SUPERVISOR] Professor Dr Yoshihito Shirai

Ezyana Kamal Bahrin
b i o ET H A N O L PhD Student, Semester 8
ezie2501@yahoo.com
CELLULASE PRODUCTION
FROM OIL PALM EMPTY FRUIT BUNCH
(OPEFB) BY BOTRYOSPHAERIA RHODINA
UPM3 IN SOLID STATE FERMENTATION (SSF)
this research is to study
the effects of SSF pa- To investigate the effect of
environmental factors on cellulase
RESEARCH OBJECTIVES
rameters that influenced
cellulase production by production in SSF
down Botryosphaeria rhodina.
a l l Botryosphaeria rhodina
produc- exhibited its best perfor- To obtain an optimized condition
tion costs. mance on day 7 of incu- of cellulase production by
Value added bation when the initial Botryosphaeria rhodina using
the solid waste moisture content was at Response Surface Methodology (RSM)
into useful products 20-25 %, initial pH of
such as organic acid, nutrient was 6 to 7 and
To study the biodegradation of
sugars, compost, biogas with 3-5 g of substrate.
and enzymes may over- Generally, fungi were
fermented OPEFB by Botryosphaeria
come the waste disposal cultivated at more than rhodina under SSF condition
▶ Botryosphaeria rhodina was
cultivated on OPEFB in solid problem in the mill. 50% of moisture content
state fermentation for cellulase Bioconversion of value in solid state fermenta- of the variables was In addition, fermented
production
added product using tion. However, high cel- preceded in response OPEFB by Botryospha-
solid state fermenta- lulase production at low surface methodology ac- eria rhodina also was
M alaysia has a well

positioned as the
major producers and ex-
tion (SSF) is economical
because it requires low
energy consumption,
moisture content (20-
25%) is a very rare con-
dition for fungi cultured
cording to Central Com-
posite Design (CCD).
Botryosphaeria rhodina
analysed according to
SEM, FTIR, TG-DTA
and compositional
porters in world’s palm low cost equipment, in solid state fermenta- exhibited its best perfor- analysis to have a bet-
oil industry. Oil palm limited water usage and tion but Botryosphaeria mance when the initial ter understanding to-
less effluent produced. rhodina was capable to moisture content was at wards the macroscopic
industry is currently
Moreover, solid state tolerate this condition. 26.3 %, initial pH of nu- observation in SSF sys-
producing the largest
fermentation condi- Response surface meth- trient was 6 and 3.95 g tem. SEM micrographs
amount of biomass in tions resemble the real od was applied in this of substrate. The model showed a remarkable
Malaysia with 85.5% out cultivation of fungi in study to improve the cel- and design on the op- fungal growth cultivated
of more than 70 million nature. Locally isolated lulase production from timization of the envi- on OPEFB for day 5 and
tonnes. In line with the fungus, Botryosphaeria OPEFB by Botryospha- ronmental factors in this 7. Cellulose and hemi-
Malaysian government rhodina is described as eria rhodina. An experi- study was dependable cellulose composition
approach to maximize an endophyte which mental design based on to predict the cellulase were gradually declined
attacks woody host. two-level factorial was production by Botry- throughout the fermen-
the use of all feedstocks,
The ascomycete fungus employed to screen the osphaeria rhodina. High tation period. However,
byproducts and waste
Botryosphaeria rhodina significant environmen- cellulase production at lignin content resided
streams, oil palm empty produces a broad range tal factors for cellulase low moisture content is in the OPEFB fiber were
fruit bunch is a poten- of lignocellulolytic en- production. From the very rare condition for remain unchanged until
tial feedstock for in- zymes such as laccases, analysis of variance fungi cultured in solid the end of the fermenta-
dustry sectors since it is pectinases, cellulases (ANOVA), initial mois- state fermentation. Thus, tion. This fact was sup-
abundant and available and xylanases. These ture content, amount the cellulase production ported with TG-DTA,
throughout the year. In- complex enzymes play of substrate and initial by this locally isolated FTIR and lignin deter-
an important role in the pH of nutrient supplied fungus required less mination which con-
tegration of Waste to
degradation process of in the SSF system were moisture and humid- clude that B. rhodina
Wealth concept is appli-
lignocellulosic materi- significantly influenced ity which render a great was unable to de-
cable to the palm oil in- als through a synergistic the cellulase production. advantage for large scale compose lignin
dustry in order to drive action. The objective of Then, the optimization production. in a week. ✳
[SUPERVISOR] Professor Dr Suraini Abd. Aziz

Zuraidah Zanirun
bioETHANOL
ieda_ring@yahoo.com
PRODUCTION OF BIOETHANOL FROM OIL PALM EMPTY

FRUIT BUNCH (OPEFB) USING CRUDE ENZYMES COCKTAIL
PRODUCED BY TRICHODERMA ASPERELLUM UPM1
AND ASPERGILLUS FUMIGATUS UPM2
A s the world larg-
est producer and
exporter of palm oil,
bioethanol production
The conversion of lig-
from biomass resources
nocellulosic materials to
as a consequence of a
bioethanol includes two
To enhance cellulases
production by T. asperrellum
RESEARCH OBJECTIVES
Malaysia shared 50% raising oil demand with
processes; UPM1 and A. fumigatus UPM2
of the palm oil produc- unstable 1. hydrolysis of cellu-
deliverance
tion and 61% of exports. lose in the lignocellulos-
prices and the agri- To optimize the production of
Palm oil industries had ic material to ferment-
cultural land competi- fermentable sugars from oil palm
able sugar and
contributed crucial in- tion for the food and
2. fermentation of empty fruit bunch using crude
come sources for the fuel supplies. Thus, the cocktail of cellulases by T. asperrellum
sugars to bioethanol.
country. Therefore the search for an alternative
Hydrolysis is usually UPM1 and A. fumigatus UPM2 using
total planted area of of biofuel sources which
catalysed by cellulases
palm oil tree increased is renewable and sub-
enzyme and the fermen-
by 4.5% to 4.69 million stitutable was in a great
tation is carried out by
hectares (http://econ. yeast or bacteria (Sun
demand. A dramatic
To develop an integrated system for
mpob.gov.my/economy/ and Cheng, 2002). In or-
increased in bioethanol the production and separation of
O ver v iew_2009.pdf). der for a hydrolysis pro-
production using food bioethanol and fermentable sugars
Thus, more wastes are cess to take part, lignin
based technology may
should be removed. Ap-
expected to be generated not be practical (Sun and
propriate pre-treatment
in the future. Oil palm Cheng, 2002). Therefore,
is essential to break up ing alternative for nol and fermentable
Empty fruit bunches lignocellulosic residues
lignin which hinders bioethanol production sugars will developed a
(OPEFB) is one of the of biomass (e.g OPEFB)
enzyme attack. With is the use of enzyme cost effective technol-
residues remained in a could be utilised as an
suitable application of as it viewed by many ogy. Process integra-
large amount, tradition- technology, OPEFB can
alternative sources for experts to be at the tion is gaining more
al practice of open burn- be fully utilised into
the production of low very cost effective in and more interest due
ing would be ended with bioethanol as an alter-
cost bioethanol. OPEFB the long term process to advantages related to
ash which then could be native of a cheaper sub-
is a lignocellulosic bio- (Hamelinck, 2005 and the reduction of energy
strates via chemical and
used as good fertiliser. mass which consists of Sun and Cheng.,2002). cost and decreased in
biological processes.
However, it was not rec- 3 major components of Enzyme cocktail was fa- the size and number of
The potential of wide
ommended as it caused cellulose, hemicelluloses
range microorganisms voured as an improved unit operation(Cardona
an environmental prob- and lignin. Hydrolysis of
utilising lignocellulosic approach to increase and Sánchez,2007). The
lem of smoked air. Yet, cellulose and hemicel-
wastes is undeniable. It the cooperative action use of a process simula-
strategy to address the involved the use of en-
luloses into fermentable of cellulases by combin- tor (SuperPro Designer:
efficient utilisation or zyme, microbes and any
sugar was the great ad- ing different sources of Intelligen, Inc., Scotch
bioconversion into vari- biological agent either
vantage for the produc- cellulases particularly Plain NJ) will greatly fa-
ous value added prod- in single and mixed.
tion of bioethanol. How- from Trichoderma and cilitate the process anal-
Hydrolysis is tradition-
ucts such as biofuel, ani- ever, complex structures Aspergillus species and ysis, including handling
ally reliant acid process,
mal feed, chemicals and of lignin make it harder thus maximising it fer- the composition of raw
however better environ-
human nutrients should for any biological pro-
ment condition favoured mentable sugars pro- material and product,
be explored deeply. cess to occur as it block-
enzymatic hydrolysis at duction. Incorporation sizing of unit operations,
Recently, much atten- ing the way of enzymes
same cost as acid hydrol- of overall processes by estimation of capital
tion has been given to attack. ysis (Hamelinck, 2005). joining together several and operating costs
Thus, one of unit operations until the (Kwiatkowski et
[SUPERVISOR] Professor Dr Suraini Abd. Aziz the promis- separation of bioetha- al.,2006). ✳

Sheril Norliana
b i o ET H A N O L Suhaimi
Master Student, Semester 5
seri_2919@yahoo.com
BIOETHANOL PRODUCTION FROM

CRUDE GLYCEROL USING
LOCALLY ISOLATED
BACTERIA
G lycerol (glycer-
ine, 1,2,3-pro-
panetriol) is highly pres-
as prom-
ence in nature since it
ising applica-
has been used in wide
tion since it may further
range of applications. add value to the biodies- ducer. The
Biodiesel production el industry. Glycerol is potential ethanol
creates abundant of believed can be used as producer obtained was tion
glycerol, since the pro- a substitute for the com- identified as Escherichia conditions
cess of biodiesel gener- mon fermentable sub- coli SS1 with ethanol such as substrate con-
ates about 10% (w/w) of strates, such as sucrose, yield of 1.0 mol/mol centration, temperature, temper-
glucose and starch, used glycerol, hydrogen as by- pH, nitrogen source, ature and trace
glycerol as the major by-
in industrial fermenta- element solution. The
product. The increased
tion processes especially effects of the significant
of biodiesel production in ethanol production as variables (two level fac-
led to increase of crude
RESEARCH OBJECTIVES
fuels. Glycerol used as torial design) were sub-

glycerol in environment, feedstock for bioethanol To isolate potential glycerol- sequently evaluated in
which consequently rise production could be an fermenting microbes for Central Composite De-
the environmental issue alternative because it is ethanol production sign (CCD) to develop
due to the problem of its cheap and abundance. an empirical model to
Hence, the studies on determine the optimum
disposal. In response to To optimize the microbial
bioconversion of crude value of each variable.
higher availability of this conversion of crude glycerin
glycerol for bioethanol The optimized condi-
compound in nature, production were perfor ethanol production by tions obtained from
many microorganisms formed by using locally statistical approach (RSM) CCD with predicted
are known to be natu- isolated bacteria. ethanol production of
rally utilizing glycerol as In this study, potential 17.04 g/L. The valida-
sole carbon and energy g lyc e rol - c ons u m i ng tion was carried out by
source. Microbial fer- bacteria that have ca- product with and minor trace element solution using crude glycerol
pability in producing amounts of organic ac- and salt content were as substrate in 1
mentation of glycerol for
ethanol from glycerol ids. This isolated strain optimized. The obtained L bioreactor. ✳
valuable chemicals for-
were isolated from envi- has greater affinity to results were subjected
mation such as 1,3-pro- ronment. The screening glycerol as compared to two-level factorial PICTURE
panediol, dihydroxy- procedure was modi- to glucose as substrate design where only four
▶ Yellow zone surrounded
acetone, ethanol and fied to isolate glycerol- for ethanol production. paramaters showed sig- bacteria colony shows the
succinate is considered consuming ethanol pro- Furthermore, fermenta- nificant result, except presence of ethanol
[SUPERVISOR] Dr Phang Lai Yee

Dayang Salwani
bt Awang Adeni
bioETHANOL
adsalwa@frst.unimas.my
THE FEASIBILITY OF SAGO HAMPAS AS A

FEEDSTOCK FOR BIOETHANOL PRODUCTION
To pretreated the sago ‘hampas’
for glucose production using
RESEARCH OBJECTIVES
enzymatic hydrolysis
S ago hampas hydro-
lysate (SHH) contain
about 85-90% (w/w) of
germinate time of com-
mercial baker’s yeast as
well as its growth profile
media containing CG.
To ensure higher etha-
nol concentration (v/v:
To study bioethanol production from
glucose after hydrolysis was observed. From >10%) which meet in-
hydrolyzed sago ‘hampas’ through
process by dextrozyme the result obtained the dustrial needs, the fer- batch fermentation system utilizing
enzyme. Higher amount yeast was not able to mentation should be commercial baker’s yeast
of glucose in SHH gives grow in the SHH me- fed with higher glucose
extra advantage as it can dia although glucose concentration. Thus to
To optimize the bioethanol
be served as a substrate was the main com- accommodate with this production from hydrolyzed sago
for bioethanol produc- ponent in the hydro- criteria three different ‘hampas’ by response surface
tion by commercial lysate. Hence, the SHH concentration (g/L: 100, method (RSM)
bakers yeast. However was then mixed with 150 and 200) glucose
SHH also contains dex- yeast extract, peptone, in SHH were initially Some by-products such sources usable by yeast
trin and maltose, as KH2PO4, MgSO4.7H2O tested. From all trials, as glycerol, lactic acid was carried out. Three
well as other inorganic and NH4SO4, the media the 100 g/L of initial glu- and acetic acid were different nitrogen sourc-
compound due to the which usually used for cose shows higher yield also produced. Higher es (NH2SO4, urea and
complex structure of culturing S. cerevisiae. with glucose has been glycerol (9 – 10g/L) was yeast extract) were stud-
hampas. Although The same media was completely consumed observed in the media of ied on their effect on
hydrolysate has been then used for control, after 16 – 18h of 150 and 200 the ethanol production,
separated from solid fermen- g/L of ethanol yield and etha-
particles, some impuri- glucose, nol productivity. Over-
ties that exist together revealed all, urea supplemented
with glucose might in- t h a t medium has shown the
fluence the capability excess capability to influence
of SHH as a susbtrate amount better ethanol produc-
for ethanol fermenta- of NADH tion compared to NH-
tion. Hence, the fer- which 2
SO4 and yeast extract in
mentation process p r o - glucose concentration of
utilizing 80 g/L of duced 150 and 200 g/L. Thus,
glucose from SHH dur- the successful of ethanol
was initially car- i n g fermentation process
ried out. All trials syn- from glucose of SHH
were conducted in 100 but for carbon thesize together with urea as
ml working volume, source it was replaced
tation process. of bio- nitrogen source has cre-
with temperature of with commercial glu-
However for 150 g/L mass and organic acids ates another economical
30°C and initial pH of cose (CG). Efficient
and 200 g/L glucose, the have been oxidized to source of C and N, for
5.5. The agitation was fermentation was then
fermentation process NAD+. production of value-
set at 100 rpm through- observed in SHH media
took more than 24h to In order to improve the added product in
out the process. On top with the theoretical con-
complete, thus affect the fermentation process our country. ✳
of that some prelimi- version yield about 98%,
volumetric productiv- utilizing high glucose
nary study on the pre- comparable with the
ity, product yield and concentration (150 and PICTURE
fermentation 200 g/L), the studies on ▶ Ethanol fermentation utilizing
[SUPERVISOR] Professor Dr Suraini Abd. Aziz e f f i c i e n c y. the effect of nitrogen sago hampas hydrolysate

Nur Amelia Azreen
b i o ET H A N O L bt Adnan
ameliaadnan88@yahoo.com
BIOETHANOL PRODUCTION FROM

GLYCEROL BY IMMOBILIZED
ESCHERICHIA COLI
I n biodiesel produc-
tion, glycerol will be
produce as secondary
energy. Bioethanol has
the ability to reduce the
greenhouse gas emis-
known for its efficiency
in improving substrate
utilization and produc-
when
compare
with free
sion by 5-7% depend tivities of various fer- suspension cells.
by-product. These phe-
on the feedstock used mentation processes. Immobilized of E.coli
nomena have become a
which is usually derived There are few advantag- is done by entrapping be transferred to 2 litre
problem since the pro- from renewable mate- es by using immobilized the cells inside sodium bioreactor to determine
duction of biodiesel is rials for example corn, cell for example it will alginate. The develop- the performance by us-
increasingly in demand. sugar beets, wheat and increase the cell den- ment of
Glycerol which is pro- wood. The availability sity, high rate of mass cell immo-
duce in the fermenta- of these raw materials transfer, intimate phase bi l i z at i on
is a major problem for mixing, high fermenta- te ch n i qu e
RESEARCH OBJECTIVES
tion will be as waste and
bioethanol production tion rate, cell recycling in regards To optimize the
problem occur where
since these raw materials and product inhibition to physical immobilization techniques
there is no way to treat
is depends on the geo- will be lower. Attention property using sodium alginate
this glycerol. These will graphic locations and has been given for fuel of alginate
be a burden to the bio- season. As a result, the ethanol production us- bead and
diesel manufacturer. cost for bioethanol pro- ing immobilized cell cell to al-
To determine the
Thus, there is a lot of induction keeps changing technology in fermenta- ginate ra- bioethanol production by
terest in the production and expensive. It is also tion processes since it tio will be immobilized Escherichia
of value added products stated by previous study has been known to show examined. coli in sodium alginate
based on glycerol such
that ethanol from glyc- a significant role in bio- The effect using 2 litre bioreactor
erol is cheaper to pro- reactor performance. of pro-
as cosmetics, paint,
duce compare to ethanol Thus the aim of this duction
pharmaceuticals, pulp from corn. Therefore by study is to enhance the media on
and paper industry and using glycerol as carbon bioethanol production the physical proper- ing bioreactor. It is ex-
also food industry. source, it can help to re- from glycerol as carbon ties of alginate beads pected that bioethanol
One of the value added duce the bioethanol pro- source by E.coli with the is important to make production from glycer-
product that been much duction cost. use of immobilized cell sure there is no leak- ol is enhanced in terms
interest is the produc- The use of immobilized system which offers ad- age of the beads during of productivity by
microbial cells for etha- vantage on the bioetha- fermentation process. using immobi-
tion of bioethanol. It
nol fermentation has nol productivity. This Cell to alginate ratio is lized cell. ✳
has become one of the
become interesting in includes investigation on optimized to maintain
most feasible options researchers due to ad- the optimum condition high cell density in the
in the area of non- vantages compare to tra- for stability and rigidity fermentation system. PICTURE
conventional sources of ditional processes. It is of the alginate beads and Once rigid and stable ▶ Washing the beads to
also the performance beads are formed, it will remove access calcium chloride
[SUPERVISOR] Dr Phang Lai Yee

Mohd Zulkhairi
Mohd Yusoff
b i o H Y D RO G E N
zulkhairi_y@biotech.upm.edu.my
FUNCTIONAL ANALYSIS OF ESSENTIAL GENE RELATED

TO BIOHYDROGEN PRODUCTION IN ESCHERICHIA COLI
B iohydrogen produc-
tion has become
pivotal subject being
an comprehensive
search of all E. coli path-
ways for their impact
glucose or formate. In
addition, all the experi-
ment has adhered with
four, and 24 hours. Me-
tabolism pathway has
been studied in response
the process become es-
sential parameters to be
investigated.
discussed recently due to on hydrogen produc- reference strain, wild to biohydrogen produc- In this study, six mu-
their eco-friendly char- tion through screening type BW 25113 [figure tion and its metabolite. tants have elucidated
acter. Its combustion the entire Keio mutant 3]. Biohydrogen amount Thus, in order to have which their genes re-
only produces H2O and library (3985 isogenic flected to biohydrogen
it is not one of the green mutants) using chemo- production. These genes
house gases. Essentially, chromic membranes have been expended for
RESEARCH OBJECTIVES
biohydrogen can be pro- (GVD Corp., Cam- further investigation
duced by various means bridge, MA) formed To screening responsible using complex formate
chemical, physical and by a thin film of WO3 genes related to biohydrogen media and two mutants
biological approaches. covered with a catalytic production in E.Coli have showed significant
Biohydrogen from fer- layer of palladium, to deviation from glucose
mentable biomass ap- detect biohydrogen gas, whereas it consumed
pears an interesting idea by a colorimetric re- To elucidate expression formate and produce
due to low cost substrate sponse [figure 1]. Several level of related genes during hydrogen with the yield
availability. Escherichia uncharacterized genes biohydrogen production. 130.05 and 80.64 µmol
coli is a robust bacterium related to bacterial hy- H2/mg protein respec-
for developmental re- drogen production were tively. The selective
search based on genetic identified and used as strains will be elected to
engineering because its candidates for further undergo molecular level
whole genome sequence investigation [figure 2]. generated in headspace better understanding investigation using qRT-
is available and its meta- The chosen strains were was analyzed using gas of the metabolism, it’s PCR in order to eluci-
bolic pathways are rela- undergone fermentation chromatography (Agi- by-products was deter- date expression level of
tively well-established. process using respec- lent Technologies Inc., mined. Carbon balance the gene in response
To date, we conducted tive substrate such as Santa Clara, CA) after or mass balance of the to hydrogenase
one, two, output and input during operon. ✳
[SUPERVISOR] Assoc. Prof. Dr. Eng. Toshinari Maeda
> FIGURE 1 > FIGURE 2 > FIGURE 3

Mohd Azwan Jenol
biohydrogen Master Student, Semester 1
azwan.jenol@gmail.com
HYVOLUTION terest in searching a new

hydrogen production
processes with almost
no carbon emission
PRODUCTION OF BIOHYDROGEN serious environmental

problems. Therefore, it
can be used as carbon
(Balat, 2009; Holladay
et al., 2009; Moriaty and
Honnery, 2009). Hy-
source for production of drogen is expected to
FROM SAGO WASTE for hydrogen production

biofuel through fermen-
tation.
Biofuels are liquid or
become one of the ma-
jor sources of energy to
replace current energy
BY LOCAL ISOLATE since biomass is cheap

and readily available.
Biomass is one of the
gaseous fuels for power
plants and transport
based fossil in the future.
As Malaysia sago palm
most abundant and re-

newable resources that
RESEARCH OBJECTIVES
yvolution is ac- can be used in order to
ronym of the in- be converted to valuable To select suitable sago waste
products. The agricul-
tegrated project “non- for biohydrogen production
tural activity is one of
thermal production of
the biggest activities that
by local isolate
pure hydrogen from
lead to the abundantly
biomass”. The concept of of biomass especially in To optimize the production of
the hyvolution is to sub- Malaysia. Biomass pro- biohydrogen from sago waste
ject the bacteria, which vides clean, renewable
using statistical analysis
is freely and efficiency in energy source that could
producing pure hydro- intentionally improve
gen as a product during the economy, environ-
ment and energy cycle.
degrading the biomass
In Sarawak, the sago mill sectors that are pro- industry produces large
(Claassen et al., 2010).
has produced significant duced from renewable amount of sago pith
The novelty of this pro- amount of post-process- resources such as bio- residue every day, there
cess is where it used bio- ing waste and residue. mass (Demirbas, 2007). has been greater interest
mass as a substrate since The final waste prod- Hydrogen is one of the in utilizing of sago resi-
there are many study has uct in the extraction of examples of biofuel that due to production
been done by wastewa- starch compound from has been found to be of biofuel such as
sago palm in the starchy well suited as it is clean hydrogen. ✳
ter. The concept of this
fibrous pith residue or and has a high calorific
hyvolution is based on
known as sago pith resi- value fuel.
combination bioprocess due (SPR). It is usually In the last few years, ▶ Experimental design for
biohydrogen production from
in exploiting thermo- washed off into drains or there are increasing in- sago waste by local isolate2
philic and phototro- nearby streams together
phic bacteria, to provide with wastewater. This
CLOSTRIDIUM
highest pure hydrogen situation will then lead BUTYRICUM EB6
production in efficiency to pollution load and
and low cost. Recently,
the production of bio-
hydrogen from biomass OPTIMIZATION OF
HYDROLYSIS OF BIOHYDROGEN BIOHYDROGEN PRODUCTION
has become a new path SAGO WASTE SAGO WASTES PRODUCTION FROM SAGO WASTE USING
STATISTICAL ANALYSIS
[SUPERVISOR]
Professor Dr Suraini Abd. Aziz
SCREENING AND ISOLATION LOCAL ISOLATION OF

OF BIOHYDROGEN PRODUCER BIOHYDROGEN PRODUCER

Siren Linggang
b i o b u ta n o l
siren_linggang@yahoo.com
PRODUCTION OF
ACETONE-BUTANOL-ETHANOL
(ABE) FROM SAGO PITH RESIDUES
T he agricultural in- fuel, the sago pith resi- UPM1 and Aspergillus
dustry produces a due needs to be treated fumigatus UPM2.
so that the cellulose and Different parameters
significant amount of
hemicellulose in the such as substrate con-
post-processing waste
plant fibers is exposed centration, temperature,
and residue. Particularly and more accessible to pH and enzyme concen-
in Sarawak, sago starch hydrolyze into simple tration was studied to
industry is responsible hexose and pentose sug- optimize the production
for the production of a ars. In this study, dried of fermentable sugars.
notable amount of resi-
due. Sago pith residue
To produce fermentable sugars
RESEARCH OBJECTIVES
is an abundant ligno-
from sago pith residue utilizing
cellulosic residue that
local microbial enzymes
left behind after starch
extraction process and
contains significant
To produce acetone-butanol- [SUPERVISOR]
amount of starch (58%),
ethanol from sago pith residues Professor Dr Suraini Abd. Aziz
cellulose (23%), hemi-
hydrolysate using Clostridium
celluloses (9.2%) and
acetobutylicum ATCC 824
lignin (3.9%). This resi-
due has a great potential
as economical substrate
for the production of
acetone-butanol-ethanol sago pith residue or sago The fermentable sugars
as an alternative fuel due hampas was ground be- obtained were utilized
to their high content of fore undergo enzymatic as a carbon source for
cellulose and hemicel- hydrolysis using dex- Clostridium acetobutyli-
trozyme. The solid part cum ATCC 824 in ac-
lulose.
(cellulose) was sepa- etone-butanol-ethanol
Biobutanol can be pro-
rated from hydrolyzed fermentation. The total
duced fermentatively sago hampas solution of acetone-butanol-
from lignocellulosic res- through filtration. The ethanol produced was
idues through ABE fer- cellulose obtained was 8.84 g/L and biobu-
mentation by clostridia used as a substrate in tanol of 5.41 g/L was
species. It is very energy this study. This cellu- achieved after 72
lose was directly con- h of fermenta-
efficient and suitable for
verted to fermentable tion. ✳
replacing petrol as fuel
sugars using cellulases
in gasoline engines. In produced by locally
conversion of lignocel- isolated fungus namely
lulosic biomass to bio- Trichoderma asperellum

Mohamad faizal
b i o B U TA N O L Bin ibrahim
mf.ibrahim@ymail.com
PRODUCTION OF ACETONE-BUTANOL-ETHANOL
FROM OIL PALM EMPTY FRUIT BUNCH
B iological production
of acetone-butanol-
existing vehicles. These
criteria of biobutanol
used; it disposed
of by mulching To produce crude cellulase
RESEARCH OBJECTIVES
ethanol (ABE) is cur- have become a great re- and dumping at
cocktail from OPEFB by
rently under demand for newable energy source plantation. In
the extraction of biobu- if the production of bu- fact, the OPEFB
T. asperellum UPM1 and
tanol. Biobutanol repre- tanol can be produce at contents high A. fumigatus UPM2
sents the next significant lower cost. amount of car- for the production of
change required to meet In the recent years, bon source in the fermentable sugar
the growth in demand growing attention has form of cellulose
for environmentally re- been devoted on the and hemicellu- To obtain ABE from
sponsible and renewable production of ABE from lose, a polymer
fermentable sugar produced
fuels for transportation. lignocellulosic biomass structure made
by Clostridium sp.
Biobutanol (C4H10O) or as an alternative renew- by sugar mono-
butyl alcohol is an alco- able energy. Oil palm mer. This poly-
hol that can be used as a tree is one of the most mer is able to
solvent or fuel produced planted for edible oil be converted into sugar ma asperellum UPM1
from biomass by a mi- in tropical countries monomers through and Aspergillus fumiga-
crobial fermentation. It such as Malaysia and hydrolysis process us- tus UPM2 using treated
has low vapor pressure, Indonesia. As a leading biological catalyst OPEFB as substrate for
can be easily blended ing industry in world’s known as cellulase. Cel- subsequently used in
with gasoline, contain oil production, palm lulase is a great product the bioconversion of
much energy as gasoline, oil industry has leaved in which works synergis- OPEFB into sugar. The
better adapted to be used behind huge amount of tic in the bioconversion sugar obtain was further
FLOW CHART DIAGRAM in the present distribu- biomass from its plan- of lignocellulosic ma- used for the production
▶ Process overview of ABE
tion system, less corro- tation and milling ac- terials into fermentable of ABE by Clostridium
production from OPEFB sive, and can be used in tivities as compared to sugars. Many technolo- sp. in anaerobic fer-
other type of agriculture gies have been devel- mentation system. The
biomass. However, its oped in order to obtain aim of this study are to
lignocellulosic residues higher efficiency of produce crude cellulase
have not been effectively hydrolysis process and cocktail from OPEFB
produce cellulase at low by T. asperellum UPM1
cost. The study was suc- and A. fumigatus UPM2
cessfully developed the for the production of
crude cellulase cocktail fermentable sugar and
produced by Trichoder- to obtain ABE from fer-
mentable sugar pro-
duced by Clostrid-
ium sp. ✳

Mohamad Nafis
Abdul Razak
b i o b u ta n o l
mohamadnafis@yahoo.com
PRODUCTION OF BIOBUTANOL FROM

OIL PALM DECANTER CAKE
T he palm oil sector
generates a larg-
which are Trichoderma
asperellum UPM1 and
exoglucanase and en-
doglucanase but lower To obtain fermentable sugar
RESEARCH OBJECTIVES
est amount of biomass, Aspergillus fumigatus in β-glucosidase activ- from pretreated oil palm
estimated at 80 million UPM2 studied prior the ity. Aspergillus sp. pro- decanter cake using enzymes
dry tonnes in 2010. In saccharification produced high activities of cocktail for biobutanol
National Biomass Strat- cess. Pretreatment of β-glucosidase can be production
egy 2020, government is the substrate is essential mix to the cellulase pro-
focusing on oil palm bi- to remove the protec- duced from Trichoderma
omass utilization to in- tive layer of lignin which sp. and produce crude To optimize production of
crease the Gross Nation- covers the cellulose and cellulase cocktail. The biobutanol from oil palm
al Income (GNI) up to hemicellulose structure. suitable ratios of both decanter cake polyoses by
RM 30 billion by 2020. This barrier reduced the cellulases make the opti- Clostridium acetobutylicum
Parallel to the initiative, efficiency of cellulase to mum activities of exog- ATCC824
production of crude hydrolyze the cellulose. lucanase, endoglucanase
cellulase cocktail, poly- Oil palm decanter cake and β-glucosidase will
ose and butanol from contains high percent- be studied to increase
oil palm decanter cake age of impurities such as the efficiency of hydrol-
(OPDC) is a great effort lignin, ash and sand. The ysis process.
to support the National effectiveness of various Biobutanol has superior
Biomass Strategy. Oil chemical and physical fuel properties when are substrate
palm decanter cake is a pretreatment (NaOH, compared to bioethanol concentra-
oil palm biomass which HCl and HNO3) to al- which become great in- tion, pH,
produced by 3-phase ter lignin content and terest to the world sci- temperature,
decanter system. In the increase the percentage entist to produce higher nitrogen
system decanter system of cellulose was studied. amount of biobutanol content and
liquid from sand cyclone The effect of pretreat- from biomass. Ace- inoculum
and the condensate of ment to the lignocel- tone-Butanol-Ethanol size. Gas
crude palm oil (CPO) lulose structure was fermentation using chromatogra-
vacuum were mixed to- observed by scanning Clostridium species is phy used to determine
gether in decanter sys- electron microscope the common method for the concentration of ▶ SEM structure
pretreated OPDC
tem and produce oil, oil (SEM). The treated bio- the production of biobu- butanol produced and
palm decanter cake and mass will proceed to sac- tanol. Optimization high performance liq-
liquid sludge. charification process by of biobutanol produc- uid chromatography
Production of crude cel- commercial enzyme and tion by using response (HPLC) used to deter-
lulases from oil palm crude cellulase. surface methodology mine the concentra-
decanter cake by lo- Trichoderma sp. pro- approach also will be tion of sugar con-
cally isolated fungus duced high amount of studied. The parameters sumption. ✳

Norhayati Ramli
bioPRODUCT PhD Student, Semester 8
eman_tihani86@yahoo.com
OVER PRODUCTION OF CYCLODEXTRIN

GLYCOSYLTRANSFERASE (CGTASE) THROUGH
MOLECULAR CLONING APPROACHES
different ratios. Thus, capability in predomi- ingly, the CGTase was
complicated and tedi- nantly producing β-CD produced extracellularly
ous purification strategy as a main product and (94%) indicating the
is needed to purify the its potential in degrad- signal peptide was func-
product. Furthermore, ing the raw starch as a tional in E. coli. In addi-
the low concentration substrate, thus known as tion, the isolation of the
of CGTase produced by a raw-starch degrading promoter and transcrip-
wild type strain caused enzyme producer. tional terminator from
the major problem for Further study on the Bacillus sp. NR5 UPM
CGTase production in cloning of a PCR gene is carried out in this
large scale. fragment from Bacillus study due to their im-
In this study, the CG- sp. NR5 UPM into an portant function in the
Tase-producing bacteria E. coli expression vector transcription process.
▶ DNA walking results for known sequences
of CGTase gene from Bacillus sp. NR5 UPM. were successfully isolat-
ed from the soil in Ma-
laysia. Out of 65, eleven To screen, isolate and characterize
RESEARCH OBJECTIVES
yclodextrin gly- Tase to convert starch CGTase producers has CGTase producing bacteria
cosyltransferase into CD brings a great been further screened
interest to research- using modified Horiko-
(CGTase, EC 2.4.1.19) is To isolate the CGTase genes by using
ers. The formation of shi agar type II with spe-
an important member of primer screening technique for
CD-inclusion complex cific indicator. The size
the amylolytic glucosy- with variety of guest of halo zones formation construction of CGTase expression
lase family that catalyzes molecules is advanta- on the indicator plate system with Escherichia coli
the formation of cyclo- geous as the enhance- is a good qualitative
dextrins (CDs) through ment of the physical and measurement of CGTase
cyclization reaction. chemical properties of producer as the biggest To evaluate the production of enzyme
The CGTase can be pro- the inclusion complex diameter formed is in- by analysing the CGTase activity of
duce by several bacterial
formed are beneficial in dicating the highest CG- the recombinant CGTases produced
biotechnology, pharma- Tase activity obtained.
species with the major
ceutical, food, cosmetic, The isolates were classi- was successfully carried The construction of E.
producers are belongs to chemical and agricul- fied using Gram staining out. Compared to the coli strain harbouring
Bacillus sp. The ability of tural field. However, the and further identifica- wild type, the CGTase pTZCGT-BS verified
CG- difficulty in producing tion was done at 16S that was produced in the importance of the
the CD in larger scale rRNA level. All eleven E.coli cells only required strong promoter in the
arises as all known isolates showed the one-fourth of culture expression of enzyme.
CGTases will characteristics of Gram time and neutral pH to The functionality of the
produce a positive and identified produce CGTase. After isolated promoter in the
mixture of as Bacillus sp. The best 12 hours of cultivation, presence of T7-vector
α-, β- and CGTase producing bac- the CGTase activity in located promoter is con-
γ-CD in teria, Bacillus sp. NR5 the culture medium firm by an increment
UPM has a probability reached approximately of 3.2-fold of enzyme
to share the biggest mar- 2.5-fold higher than activity compared
ket in the industrial ap- the CGTase from the to the parent
plication since it has the parental strain. Interest- strain. ✳

▶ Screening of CGTase-
producing bacteria with
61
Horikoshi + phenolphthalein
RESEARCH REPORT 2011 > EB GROUP

Rozaimi Abu Samah
b i o P RO D U CT
rozaimiabusamah@gmail.com
DEVELOPMENT OF A RECOVERY
SYSTEM FOR BIOVANILLIN
V anillin is one of the
mostly used fla-
vors all over the world.
The bioconversion of
several substrates to
vanillin normally pro-
fruit bunch (OPEFB).
Subsequently, the fer-
mentation broth will
To adsorb vanillin using
adsorbent resin
RESEARCH OBJECTIVES
It is normally found duces relatively low undergo two different
in the bean of Vanilla yields. However, one techniques in recover-
To recover vanillin from
planifolia (Reineccius, of the mentioned sub- ing biovanillin, which fermentation broth via
2006). Presently, it is strate, ferulic acid, can are cross-flow filtration crossflow filtration
produced synthetically be considered as the and membrane-based
to meet the market de- most promising sub- solvent extraction. Both To recover vanillin from
mands. However, only strate in the production techniques will be evalu- fermentation broth using
0.2% is extracted from of biovanillin. Though, ated mainly in term of membrane-based solvent
beans (Priefert et al., it has its own drawback, its efficiency. extraction
2001). Due to these fac- in which at certain con- It continues with puri-
tors, extensive studies centration of vanillin, it fication stage involving To evaluate the economic
are carried out on the inhibits the enzymatic macroporous adsorp- feasibility for vanillin recovery
production of vanillin reaction as well as be ox- tion resin. Several res- from fermentation broth
via microbial transfor- idized further to vanillic ins are tested for their
mation from different acid (Wu et al., 2008). capabilities of adsorbing
substrates such as lignin Therefore, it is an advan- vanillin from aqueous
(white rot fungi), phe- tage to produce biovan- samples. Adsorption
nolic stilbenes (Pseu-
domonas paucimobilis),
illin using an integrated
bioprocess system which
parameters such as con-
tact time, solute initial
O H
ferulic acid (Bacillus comprises of both up- concentration, pH, tem-
coagulans), and eugenol stream and downstream perature and amount
(Corynebacterium sp.). sections. of resins is screened
Other than that, enzy- The first section of this and optimized in order
matic routes also yield
an acceptable amount
work is biovanillin pro-
duction via fermenta-
to obtain the highest
amount of biovanillin. CH3
of vanillin such as di-
oxygenase reaction on
tion using Pycnoporus
cinnabarinus. One at-
Lastly, economic assess-
ment for each separa-
O
isoeugenol (Yoshimoto
et al., 1990), soybean li-
tractive point in this
research is that the sub-
tion route will be done.
Separation cost and OH
poxygenase reaction on strate, ferulic acid, will purification yield will
eugenol (Wu et al., 2008) be obtained from one be the pivotal points in
and beta-glucosidase re- of the most abundant deciding the best sep-
▶ Vanillin structure
action on glucovanillin by-products in Malay- aration route for
(Dignum et al., 2001). sia, the oil palm empty biovanillin. ✳

Nurul Asyifah
bioPRODUCT Mustapha
syifah86@yahoo.com
ENZYMATIC IMPROVEMENT OF
NUTRITIONAL VALUE OF BROWN RICE
FOR POULTRY
T he utilization of lo-
cal feedstuff which
ence of anti-nutrients or
non-starch polysaccha-
brown rice since poul-
try cannot produce en-
To determine the nutrients and
RESEARCH OBJECTIVES
is brown rice for poultry rides in brown rice has dogenous enzymes to anti-nutritional factors (mainly
feed has been studied reduced the availability degrade these NSP. The NSP) in brown rice
using few varieties. The of other nutrients to be enzymes addition can
varieties of MR239 and absorbed by poultry. supplement or help the To optimize the enzymatic hydrolysis
MR257 showed that The fibre content, cellu- endogenous digestive conditions using Response Surface
their nutrients are not lose, hemicellulose, ara- activities of poultry, re- Methodology (RSM) for production of
preferred for human binoxylan and beta-glu- move anti-nutritional hydrolyzed brown rice
consumption but can be can are the components factors and also render
obtained in high yield. that were determined certain nutrients more
To evaluate the effect of enzyme
The energy and nutrient as non-starch polysac- readily available for ab-
composition of brown charides in brown rice. sorption and enhance
addition on poultry
rice are similar with the Even though these non- the energy value. The
maize which is the main starch polysaccharides optimization using Re-
feed used in Malaysia. presence in low percent- sponse Surface Meth- ratory scale to produce true metabolisable energy
Thus, these two varie- age, a suitable treatment odology (RSM) for en- hydrolyzed brown rice. (TME) value and com-
ties of brown rice has should be taken to re- zymatic hydrolysis of This hydrolyzed brown pared it with rice with-
been analyzed for their move it. brown rice using com- rice was force f-ed to out enzymes supple-
proximate composi- The addition of enzymes mercial enzymes was chicken to determine the mented. ✳
tion to determine the has been widely used to done in labo-
protein content, fat, en- remove the non-starch
ergy value and also the polysac-
fibre content. The data cha-
obtained from varie- rides
ties MR239 and MR257 a n d
has supported that local to im-
brown rice can replace prove
maize all together or at the nu-
certain rate for poultry tritional
feed. However, the pres- value of
▶ Chickens for force feeding
[SUPERVISOR]
Professor Dr Suraini Abd. Aziz

Fairouz Jahaan
Mohd Aanifah
bioPRODUCT
fai1387@yahoo.com
BIOVANILLIN PRODUCTION
FROM ALKALINE HYDROLYSATE OF
OIL PALM EMPTY FRUIT BUNCH (EFB)
P alm oil industry in vanilla flavour, could ferulic acid to lignin and
Malaysia has been a also be generated from hemicelluloses. A few
OPEFB fibre, where this treatment conditions are
great strength over half
actually meets the re- tried to achieve the best
centuries, especially in
quirement and demand ferulic acid release from
term of economic de- by the consumers on OPEFB. The alkaline
velopment. As oil palm natural flavours, impor- hydrolysate containing where resin is used to
plantations cover a ma- tantly to substitute the ferulic acid as the pre- trap the vanillin and
jority of planting land, synthetic flavour used cursor was fermented avoid further conver-
the vast production sion to vanillyl alcohol.
and processing result in An optimization study
will also be carried out
abundance of by-prod- To obtain alkaline hydrolysate
RESEARCH OBJECTIVES
to obtain the optimal

ucts. Oil palm empty of oil palm empty fruit bunch
conditions of biovanillin
fruit bunch (OPEFB) containing ferulic acid production. The physi- PICTURES [FROM TOP]
is the major palm oil cal aspects to be opti- ▶▶ Ground oil palm empty
industry’s waste. Agri- fruit bunch
To obtain biovanillin by mized comprise of the ▶▶ A. niger ATCC 6275
cultural wastes contain optimizing ferulic acid following parameters; ▶▶ A. niger ATCC 6275 in alka-
temperature, initial pH, line hydrolysate of OPEFB
ferulic acid, a hydroxy- bioconversion to biovanillin ▶▶ Locally isolated P. chrys-
cinnamic acid of the agitation speed and ini- osporium
through two-stage fungal tial inoculum concen-
phenolic group that can fermentation using 2-level tration. An optimized
be used as a biologi- factorial design condition of shake flask
cal precursor to be fer- fermentation to produce
mented into biovanil- biovanillin shall be
lin. Varied biomass had achieved at the end
been utilized before to in food and beverages through two-stage fun- of this study. ✳
generate biovanillin like industries. OPEFB was gal fermentation to pro-
rice bran oil, maize bran, alkaline treated to reduce biovanillin. The
lease ferulic acid. This OPEFB hydrolysate was
corn cobs, sugar beet
is because alkaline treat- converted into vanillic
pulp and others through
ment is well known for acid by Aspergillus niger
biotechnological route. its delignification effi- ATCC 6275 and further
In that case, vanillin, ciency and breaking up into biovanillin by Phan-
the active ingredient of ester bonds attaching erochaete chrysosporium,

Nur Ain Zamzuri
bioPRODUCT PhD Student, Semester 2
aenzamzuri@yahoo.com.my
ONE-STEP BIOTRANSFORMATION
OF FERULIC ACID INTO BIOVANILLIN
USING GENETICALLY ENGINEERED
ESCHERICHIA COLI To screen, isolate and
RESEARCH OBJECTIVES
identify potential biovanillin
producing bacteria
To isolate the functional genes for

illin extracted from va- biotransformation of ferulic acid into
nilla pod as well as the
biovanillin and further construct
chemically synthesize
chemically synthesized vanillin in the near fu-
genetically engineered E.coli
vanillin. However, this ture. Microbial or en-
To produce biovanillin
artificially derived van- zymatic transformation
in one step fermentation using
V anillin is the ma- illin flavour could not route has been reported
jor component be referred as a natural as the most promising
of natural vanilla and product. way to produce vanillin
genetically engineered E.coli
a secondary metabo- Therefore, the recent from the precursors like

lite of plant which is increasing demand for ferulic acid, vanillic acid, from ferulic acid, the study, bacteria named as
an important aromatic natural flavours and the eugenol or isoeugenol. main concern is on the Pseudomonas sp. AZ10
component as well as problem of vanillin de- The bacterial biotrans- vanillin further oxida- UPM has been success-
flavouring compound rived from Vanilla plant formation pathway was tion into vanillic acid fully isolated as a poten-
in the industry of food is relatively expensive compared in order to which resulted to the tial biovanillin producer
and personal products. has move the trends to- screen and select the poor yield of vanillin. using ferulic acid as sole
It is derived from the wards investigation of functional genes for bio- Based on the common carbon source. By us-
tropical Vanilla orchid other biotechnological vanillin to be produced pathway of bacteria for ing this strain, isolation
by the extraction from routes to produce van- efficiently. biovanillin production, of functional genes for
vanilla beans. Natural illin. As a result, vanil- In this study, ferulic acid vanillin will be further biovanillin production
vanillin extracted from lin production through will be used as biovanil- oxidized into vanillic can be carried out using
vanilla pods has a very biotransformation of lin precursor due to the acid due to oxidation of DNA walking strategy
high price and limited potential precursor by chemically close rela- vanillin was easily oc- and later can be cloned
supply in the market due microorganism has tionship to vanillin, low curred in compared to and expressed into
to it involved a time- been proposed towards cost, and readily availa- ferulic acid. As a result, pUC19 vector. The con-
consuming process a sustainable and en- ble. It has been reported vanillin as an intermedi- struction of genetically
which required intensive vironmental friendly that the bacterium from ate was nearly undetect- engineered E.coli con-
cultivation, pollination, process. The researchers ‘Pseudomonas’ family able at the end of the fer- taining biovanillin func-
harvesting and ripening are now investigating have the ability to pro- mentation process. tional genes is expected
of pods. It is also very the potential biovanillin duce vanillin via bio- Thus, the aim of this to produce biovanillin
dependable on the suit- production through biotransformation process study is to develop a in one step fermentation
ability of soil and climate technological approach involving ferulic acid methodology for bio- without further oxida-
conditions. Thus, cur- which can be regarded obtained from biomass. vanillin production tion of vanillin into
rent market demand for as natural product to Through the investiga- using genetically engi- vanillic acid. ✳
vanillin is fulfilled by the replace the natural van- tion of biotechnologi- neered E.coli by one step
cal routes for pathway without further PICTURE
biovanillin oxidation of vanillin into
▶ Isolated bacteria
[SUPERVISOR] Professor Dr Suraini Abd. Aziz production vanillic acid. From this (Pseudomonas sp. AZ10 UPM)

BIOMATERIAL
RESEARCH GROUP
”Utilization of biomass for the production of bioplastics,
biocomposites and organic acids”
 RESEARCH SYNOPSIS
O
ur main research ids. These organic acids (PP) and bioplastics. We is being taken into ac- squeezing. We currently
activity is to can be used as the sub- are also developing PHA count. We introduce an utilize the OPF juice for
utilize biomass strates to produce bio- blends whereby PHA eco-friendly process for the production of PHA
for the production of degradable plastics, i.e. is blended with other the recovery of PHA and organic acids. Due
biomaterials such as polyhydroxyalkanoates thermoplastic polymers from the cells by using to its potential to be used
bioplastics and biocom- (PHAs) and also for the such as polyethylene water as solvent, and we as substrate for the pro-
posites, and biobased production of green (PE) and poly(methyl also introduce the chem- duction of bioplastics,
chemicals such as or- solvent which is ethyl methacrylate) (PMMA). ical recycling concept organic acids, biofuels
ganic acids and its es- lactate. Biocomposites Production of crotonic for the PHA bioplastics. and biobased chemicals,
ters. Up to date, we have from palm biomass are acid is also of our in- Recently, our group has the Malaysian govern-
successfully utilized currently under devel- terest whereby the acid successfully demon- ment has included OPF
palm oil mill effluent opment. We utilize oil can be obtained by the strated that OPF petiole juice pathway as a route
(POME) and municipal palm mesocarp fiber, oil pyrolysis of PHA. Apart can be a good source to the production of
solid waste (MSW) fresh palm empty fruit bunch from the development of of fermentable sugars biobased products, and
leachate for the produc- (EFB) and oil palm biomaterials, our focus is (glucose, fructose and this has been announced
tion of organic acids; frond (OPF) petiole fib- also on the downstream sucrose) whereby the during the launching of
namely acetic, butyric, er to reinforce polymers processing whereby the sugars can be extracted National Biomass Strat-
propionic and lactic ac- such as polypropylene environmental concern from the OPF petiole by egy 2020, recently. ✳
▶ Elmy placing the sample tube in the 500- ▶ SEM images of Chloroform extracted PHBV
▶ Bioplastic pilot plant at SIRIM, Shah Alam
MHz JEOL JNM-ECP500 FT NMR system granules from Comamonas sp. EB172

 RESEARCH AREAS
STRAIN IMPROVEMENT
FOR PHA PRODUCTION
Genetic engineering techniques are utilized in order to improve
our locally isolated bacterium, Comamonas sp. EB172. We expect
to develop a recombinant bacterium with higher capability of PHA
production in comparison to the wild type strain.
Upstream
Downstream Processing
Processing 1. DEVELOPMENT OF BIOMASS
1. PHA recovery This includes the preparation of biomass prior to the production
of biobased products, for example squeezing the OPF petiole for
An environmental friendly method for the recovery of PHA is used obtaining the sugary juice and steam treatment of palm fibers for
whereby the PHA is extracted from the bacterial cells with the use the production of biocomposites.
of non-halogenated aqueous solvents such as low concentration
NaOH and water.
2. Production of PHA, organic
2. Chemical recycling of PHA acids and ethyl lactate
Chemical recycling is aimed at converting the PHAs into mono- Fermentation is done to produce PHA and organic acids. Optimi-
mers or low molecular weight polymers which can be reused to zation of fermentation parameters is done in order to gain maxi-
produce new polymer materials. So far we have tried two methods mum yield. Ethyl lactate is produced by esterification of lactic acid
for the depolymerization of PHA, i.e pyrolysis and hydrolysis. with ethanol.
3. Recovery of crotonic acid 3. Biocomposites and PHA blends

from PHA-producing bacteria
Biocomposites are prepared from treated palm fiber and thermo-
Crotonic acid can be obtained by thermal degradation of PHA un- plastic polymers such as PP and PHA by melt-blending. The effect
der controlled temperature and retention time. Crotonic acid and of compatibilizer on the properties of biocomposites is also studied.
its derivatives can be used in cosmetics, plasticizers, paints, etc. PHA blends are prepared by blending PHA with PE and PMMA.
▶ Comamonas EB172 culture ▶ Bioreactor 2 liters for production PHBV ▶ POME and clarified acid

BIOMATERIAL
RESEARCH GROUP
SUBSTRATE
PROFESSOR DR DR HIDAYAH
MOHD ALI HASSAN ARIFFIN
Treated Kitchen Oil Palm
POME Waste Fronds PRINCIPAL RESEARCHERS
Treated POME
Syaiful Acid Recovery Elmy recycling of pha Mohd

Nizam Nahida Nor Faiz
strain
improvement
Yee
Cupriavidus Necator
CCUG52238
Mior production of pha phbv recovery

Ahmad Nor Asma Mitra
Khushairi
media / culture
optimization
Azman
crotonic acid
Mohd Noor Ida biocomposite Mohd
production
Rahimi Amalina Nor Faiz

THE FIRST MALAYSIAN
BIOPLASTIC PILOT PLANT
The launching of the 1st Bioplastic Pilot Plant in

Malaysia by the Minister of Science, Technology
and Innovation in July 2011 was our biggest
achievement in 2011. The pilot plant was developed
under TechnoFund project, a government funded
research project. Apart from UPM, other institutions
involved in the research are SIRIM, USM and MIT.
The research project led by SIRIM was aimed at
producing PHA from palm oil and its derivatives at
large scale. With the experts from the institutions
involved, the team has successfully developed a
pilot scale fermentation and recovery system which
consists of four bioreactors, with capacity of 20, 200 (2 units) and 2000 L. The pilot plant was built
under the supervision of local experts from SIRIM, using local technologies. The interesting part in
this PHA pilot plant is the PHA recovery system which does not use the conventional halogenated
organic solvent. Under this project, we managed to introduce an eco-friendly PHA recovery system
which uses mild alkali. The pilot plant is located at SIRIM, Seksyen 15, Shah Alam. 
▶ PHA recovery system using ▶ Left to Right: 20, 200 and 2000L
non-halogenated solvent bioreactors at bioplastic pilot plant

Syaiful Nizam
Bin Basri
b i o M AT E R I A L
synba_5143@yahoo.com
ANAEROBIC TREATMENT OF PALM OIL MILL

EFFLUENT FOR ORGANIC ACIDS PRODUCTION
IN PILOT SCALE BIOREACTOR
M alaysia is one of major sources, which of POME are volatile To investigate the effect
RESEARCH OBJECTIVES
the largest coun- is sterilizer condensate, fatty acids that mainly
of hydraulic retention
tries that involved in hydrocyclone waste and consist of acetic, propi-
separator sludge. POME onic and butyric acids.
time, pH and agitation
palm oil industry. The speed on organic acids
is thick brownish liquid These acids then will be
total production of palm production at pilot scale
which high nutrient used as a substrate for
oil in Malaysia contrib- content mainly oil and polyhydroxylakanoate
uted 45% of the palm fatty acids. This nutri- (PHA) production. To study the recovery
oil demand. With such ent content is able to In this research, the or- efficiency of organic
a huge production, the support bacteria growth ganic acids are produce acids in pilot scale rotary
palm oil industry gen- with the degradation in pilot scale acidogenic evaporator system
erates large amounts of of the waste to reduce bioreactor. Hydraulic
its pollution strength. retention time, pH and
wastewater known as
During the biological agitation speed were
palm oil mill effluent
treatment of POME, by control in order to ob-
(POME). POME is gen- products recovered dur- tained highest produc-
erated mainly from three ing anaerobic process tion of acids. Organic acids produced then PHA has limited the use
will be recovered in pilot of these biodegradable
[SUPERVISOR] Professor Dr Mohd Ali Hassan scale rotary evapora- plastic for the time be-
tion system. With large ing. The production of
amount of wastewater PHA based on organic
generate in palm oil acids as carbon sources
industry each year, the will be useful to reduce
raw material for organic the price and depend-
acids production will be ing on synthetic carbon
continuously exist. PHA sources only. Therefore,
is a biopolymer and bio- this study will give sig-
degradable thermoplas- nificant contribution to
tics that have been pro- new knowledge aside
duced by various types improve efficiency of
of bacteria as carbon POME treatment, reduc-
and energy reserve ma- ing the cost and enhance
terials. The high price of the environment. ✳
PICTURE (LEFT) PICTURE (BACKGROUND)
▶▶ Pilot scale rotary ▶ Pilot scale

evaporator (50 l) acidogenic bioreactor

Mohd Rahimi Zakaria
b i o m at er i a l @ Mamat
pioneer_asis01@yahoo.com
Alternative route to crotonic acid production

by pyrolysis of polyhydroxybutyrate (PHB)-
containing bacterium
C rotonic acid is a
short chain unsatu-
This is accompanied
by about 1500m3 of
polyhydroxybutyrate
(PHB)-producing bac-
To propose an alternative method for
RESEARCH OBJECTIVES
rated carboxylic acid. contaminated effluent terium. This bacterium
crotonic acid production via pyrolysis of
crotonic acid and its per ton of crotonic acid accumulates PHB as
derivatives have various from the drying process. energy reserve mate- polyhydroxybutyrate (PHB)-
specific applications; for Moreover, the crystal- rials under suitable containing bacterium
example as a compo- lization process also conditions during fer-
To optimize pyrolysis conditions for
nent in dental materials, causes product losses. mentation. PHB can
cosmetics, hair styling The present proposed later be converted to its enhanced recovery of crotonic acid
products, plasticizers, research provides an dehydrated monomer from the PHB-containing bacterium
herbicides, compatibi- alter- that is crotonic acid via
lizers, paints thermal degradation. In To analyze, quantify and determine the
and hydro- this research, recovery purity and geometric isomerism of the
gels. Current of crotonic acid from crotonic acidproduced from pyrolysis
production of PHB- producing bacte-
crotonic acid rium will be conducted
is via chemi- by the mean of pyrolysis.
cal synthesis; Pyrolysis of PHB- con-
however it taining bacterium will
has several be conducted in a glass
drawbacks. tube oven, and the py-
The chemi- rolzates will be collect-
cal synthesis ed and characterized
of crotonic by 1H-NMR, GC and
acid involves HPLC. The geometric
many steps. isomerism of crotonic
Fu r t he r more, acid produced will be
purification of determined by 1H-
crotonic acid by NMR. This study is
crystallization expected to contrib-
may contribute natives ute to the proposal of al-
to the environmental route to crotonic acid ternative route for croy-
pollution as it causes the production which in- tonic acid production
formation of about one volves biological syn- which is more environ- PICTURE (LEFT) PICTURE (ABOVE)
ton of highly contami- thesis and eco-friendly mental friendly and
▶ Freeze-dried cell ▶ Glass tube oven for
nated effluent per ton of method. This can be involves less pro- containing PHB pyrolysis experiment
processed crotonic acid. done with the use of duction steps. ✳
[SUPERVISOR] Dr Hidayah Ariffin

Yee Lian Ngit
B I O M AT E R I A L
lianngityee@hotmail.com
PRODUCTION OF POLYHYDROXYALKANOATES BY
GENETIC ENGINEERED ESCHERICHIA COLI JM109
C omamonas sp. EB
172 has been identi-
fied as a novel bacterium
the GenBank online da-
tabase under accession
numbers HM853676,
of 246 amino acids with
a calculated molecular
mass of 26.27 kDa.
polymerised P(3HB) tion for the best growth
when fed with glucose and accumulation in
and volatile mixed or- batch fermentation us-
that is able to convert JF773394, HQ650140 The PHA biosynthe- ganic acids (acetic acid: ing 2 L bioreactor. The
organic acids derived and HQ650141, respec- sis genes (phaCCo and propionic acid: n-butyr- higher concentration
from palm oil mill efflu- tively. Sequence analysis phaABCo) were success- ic acid) in ration of 3:1:1, of glucose (20 g/L) had
ent, into bioplastics. In of the phaACo, phaBCo fully cloned in a heterol- respectively. The E. coli increased the maximum
my study, PHA depoly- and phaCCo genes re- ogous host, Escherichia JM109 transformant cell concentration of
merisation gene (phaZCo vealed that they shared coli JM109. E. coli JM109 harbouring phaCABCo the recombinant E. coli.
gene, 1333 bp) and more than 85%, 89% and transformants harbour- could accumulate However, the PHA yield
biosynthesis operon of 69% identity, respec- ing pGEM’-phaCCoABRe P(3HB) at 2 g/L of pro- based on the substrate
Comamonas sp. EB172 tively, with orthologs and pGEM’-phaCReABCo pionic acid. P(3HB) utilised coefficient (g
consisting of three genes from Delftia acidovorans were shown to be func- contents of 40.9% and PHA/g substrate) and
encoding acetyl-CoA SPH-1 and Acidovorax tionally active synthesiz- 43.6% were achieved by PHA biosynthesis per
acetyltransferase (phaA- ebreus TPSY. The phaCCo ing 33 wt% and 17 wt% using 1% of glucose and cell (g PHA/g cell) were
Co
gene, 1182 bp), ace- gene encoded a protein of poly(3-hydroxybu- mixed organic acids, re- similar using lower
toacetyl-CoA reductase of 564 amino acids, and tyrate) [P(3HB)]. E. coli spectively. amount of glucose. The
(phaBCo gene, 738 bp) the calculated molecular JM109 transformant Unlike shake flask fer- increasing of carbon
and PHA synthase, class mass was 62.94 kDa. The harbouring the three mentation, pH was sources had increased
I (phaCCo gene, 1694 phaACo gene encoded genes from the acid- maintained using NaOH the productivity
bp) were identified. The a protein of 394 amino tolerant Comamonas (1M) and DO level can of the PHA. ✳
nucleotide sequences of acids with a calculated sp. EB172 (phaCABCo) be main-
phaZCo, phaCCo, phaACo molecular mass of 40.73 under the control of na- tained at
and phaBCo reported kDa, while the phaBCo tive promoter from Cu- 30% us- To determine the PHA
RESEARCH OBJECTIVES
here were deposited in gene encoded a protein priavidus necator, in vivo ing agita- biosynthesis genes from
Comamonas sp. EB172
To study heterologous expression
of phaAB and phaC genes
from Comamonas sp. EB172 in
Escherichia coli JM109
784 bp 1333 bp 659 bp
To determine PHA production by
using constructed recombinant
Escherichia coli JM109
phaZ RnfABCDGE
yjbN
PHA Depolymerase Electron transport
Protein
intracellular type B ▶▶ Organization of Comamonas sp.
EB172 PHA depolymerase gene

Mior Ahmad Khushairi
b i o m at er i a l Mohd Zahari
ahmadkhushairi@gmail.com
PRODUCTION OF POLY(3-HYDROXYBUTYRATE)
FROM OIL PALM FROND
P oly(3-hydroxybu-
tyrate) (PHB) is a
biodegradable thermo-
supplies a renewable
biomass which can be
further utilized to pro-
forth. Further pretreat-
ment by centrifugation,
membrane filtration and
To determine the appropriate
plastic polyester accu- duce other value added sterilization will be em-
mulated intracellularly product such as bio- ployed to optimize the sugar extraction and pretreatment
by many microorgan- plastic. Based on these fermentable sugars pro- method from oil palm biomass
RESEARCH OBJECTIVES
isms under unfavorable findings, an attempt has duction. To optimize poly(3-
growth conditions. In been made to ferment The next step of this
hydroxybutyrate) [P(3HB)]
PHB production, about sap from oil palm bio- study will be bioplastic
40% of the total produc- mass to produce PHB by fermentation, the stage
production by using sugar from
tion cost is for raw ma- using bacteria. in conversion of fer- oil palm biomass in shake flasks
terial. Thus, the use of a This research will be mentable sugar to PHB
cheaper carbon source divided into three dif- by wild type of bacteria To optimize the production of
is required in order to ferent stages which are C. necator CCUG52238, P(3HB) in 2-L bioreactor
reduce the high produc- extraction of sugar from and the mutant of C. To scale-up the fermentation
tion cost of PHB. Palm oil palm fronds (OPF), necator NCIMB11599.
process from 2L to 50L
oil industry in Malaysia followed by, PHB pro- During fermentation,
has contributed about duction through fer-
bioreactor and to optimize
several experiments
mentation in bioreactor will be carried out to the process with respect to the
and finally, extraction, optimize the PHB pro- process parameters
▶ Poly(3-nydroxybutyrate) purification and charac- duction in shake flasks.
from oil palm frond juice
terization of PHB from These were include;
the cell. Prior to effect of substrate con-
fermentation, centration, effect of tem-
juice from perature, effect of agita-
OPF will be tion and effect of initial eters in the fermentation C/N ratio and tempera-
extracted pH will be observed by process have been opti- ture will be optimized to
by using 500ml flasks as the mized, the 2-L bioreac- increase CDW and PHB
simple phys- fermentation system. tor system will be used yield. The conventional
ical separation During optimization in as a tool to observe the method of extracting
method. At this shake flasks, the pro- potential of scaling up PHB from fermentation
52% of stage, characterizations file of various physical to 50-L bioreactor of the broth by chloroform and
the total world oils and of OPF juice will also parameters such as cell bioplastic production evaporation will then be
fats export in year 2006. been carried out. These dry weight (CDW), PHB from oil palm biomass. applied to observe the
Apart of its contribution were including sugar concentration and PHB At this stage, several PHB characteristic
to economic growth, composition, proxi- content will also being parameters such as stir- produced from oil
palm oil industry also mate analysis, and so studied. After param- ring speed, aeration, pH, palm biomass. ✳

Noor Azman
Mohd Johar
noorazman23@yahoo.com
Optimization of polyhydroxyalkanoates
production by an acid-tolerant
bacterium, Comamonas sp. EB 172 using
M ixed organic ac- for producing polyhy- 10% (v/v)), concentra-

Optimization for production of
RESEARCH OBJECTIVES
ids derived from doxyalkanoates (PHAs). tion of (NH4)2SO4 (0-1
anaerobically treated Statistical approach, g/l) and concentration Polyhydroxyalkanoate (PHA)
palm oil mill effluent central composite de- of mixed organic acids by Comamonas EB172 using
(POME) containing sign (CCD) was used (5-10 g/l). The analysis Response Surface Methodology
acetic:propionic:butyric to investigate the com- of variance (ANOVA) (RSM)
(ratio of 3:1:1) were plex interaction among showed that all of these
used as carbon source temperature (25-37 oC), five factors were signifi- Develop a simple kinetic
in the batch culture of initial medium pH (5- cantly important in the
model for production of PHA
Comamonas sp. EB172 9), inoculum size (4- batch fermentation by
shake flask with the P
in 2 liters bioreactor
value less than 0.001.
The optimal tem-
perature, initial me-
dium pH, inoculum flask fermentation, pre- neous considerations of
size, concentration diction for the growth substrate inhibition, cell
of (NH4)2SO4 and was at 2.98 g/l of dry growth, maintenance
concentration of cell weight (DCW) with and product formation
mixed organic ac- 47.07 wt.% of PHA con- were explored. Results
ids were determined tent. The highest yield of showed that growth
at 30oC, 7.04, 4.0% PHA was 0.28 g of PHA of of Comamonas sp.
(v/v), 0.01 g/l and per g mixed organic ac- EB172 was inhibited
5.05 g/l respectively. ids. For the second ob- under initial-sufficient
Optimization of the jective for this study was conditions. From the
production medium to determine the kinetic experimental results
containing mixed or- of the PHA production verify that the model
ganic acids improved and the mixed organic established in this work
the PHA production acids consumption in was able to describe the
more than 2 fold. the 2 L bioreactor with PHA production from
Under optimal con- the optimal condition mixed organic acids
dition in the shake obtained from the first by Comamonas
objective with simulta- sp. EB172. ✳
▶ Fermentation in bioreactor for PHA production

Mitra Mohammadi
b i o m at er i a l PhD Student, Semester 7
mitramohammadi@gmail.com
ALTERNATIVE RECOVERY METHODS OF

INTRACELLULAR POLYHYDROXYALKANOATES
FROM LOCAL ISOLATE COMAMONAS SP. EB172
To evaluate the use of NaOH
RESEARCH OBJECTIVES
treatment for PHA recovery from
local isolate Comamonas sp. EB172
To evaluate the use of chemical-
free aqueous solution method for
PHA recovery from local isolate
Comamonas sp. EB172
Comamonas sp. EB172 Recombinant Cupriavidus necator
To characterize the PHA recovered
▶▶ Thickness of bacteria cell wall from local isolate Comamonas sp.
EB172 using NaOH treatment and
chemical-free aqueous solution method
I n this study, the use of

NaOH treatment and
chemical-free aqueous
recombinant Cupriavi-
30°C for 5 h could effi-
dus necator as control.
ciently extract the PHA
The PHA recovered un-
with more than 93%
ence in the thickness
of their cell membrane.
the mild NaOH treat-
ment and the chemical-
solution for intracellular der the most favourable
of recovery yield. Pro- The initial intracel- free aqueous solution
polyhydroxyalkanoates conditions was further
tein determination and lular PHA content methods developed can
(PHAs) recovery from characterized.
transmission electron also affected the effec- serve as alternative re-
local isolate Gram-neg- The chemical-free aque-
microscopy images as tiveness of the extrac- covery methods with
ative wild type bacteria ous solution method
well as gas chromatog- tion methods for PHA high potential instead
Comamonas sp. EB172 as a clean process and
raphy analysis proved recovery. Although, the of the conventional
at different NaOH con- low concentration of
that improvement in cell NaOH treatment could halogenated solvent ex-
centration, treatment NaOH treatment was
wall permeability and recover purer PHA as traction process such
time, temperature and successfully devel-
cell membrane break- compared to the chem- as chloroform, since
agitation is being evalu- oped for PHA extrac-
age are the possible ical-free method, both these new methods are
ated as alternative and tion from wild type
mechanisms of NaOH processes were able to environmentally more
appropriate methods Comamonas sp. EB172.
treatment and chemical- extract the polymer with benign, effective and
instead of halogenated More than 88% purity
free aqueous solution high yield. The chem- simple in operation. The
solvent extraction sys- and 96% recovery yield
method on PHA recov- ical-free aqueous solu- recovery of intracellular
tem. These methods of PHA were achieved
ery which was accom- tion method was found PHA from Comamonas
consist of recovery steps by incubating the wild
panied by considerable to be better than NaOH sp. EB172 cells via the
followed by incubation, type bacteria under the
release of protein after treatment for PHA ex- recovery methods de-
centrifugation and puri- mild alkaline treatment
the extraction step. It traction in respect of the veloped herein can con-
fication steps using etha- using 0.05 M NaOH at
was found that the effec- final polymer molecular tribute towards the sus-
nol and water washing. 4°C for 1 h. The incuba-
tiveness of chemical and weight, which is in fact tainable process of PHA
The results of this study tion of Comamonas sp.
non-chemical treat- almost double that of the production using organ-
were compared with EB172 in the chemical-
ments depends on the chloroform-extracted ic acids derived from the
PHA extraction from free aqueous solution at
microbial strain which PHA as control. anaerobic treatment of
might be due The overall results in palm oil mill efflu-
[SUPERVISOR] Professor Dr Mohd Ali Hassan to the differ- this study indicated that ent (POME). ✳

Nor Asma Ab Razak
B I O M AT E R I A L
asmaktdi2@yahoo.com
IMPROVED RECOVERY AND PURIFICATION

OF PHA (POLYHYDROXYALKANOATES)
FROM RENEWABLE RESOURCES
ing the challenge es for the complete PHA range of properties. The
of global climate recovery. PHA from Co- micrograph, chemical,
change. PHA mamonas sp. will be re- mechanical and thermal
are synthesised covered by using meth- properties are investi-
when bacteria ods sodium
are exposed to a hydroxide,
To develop an alternative,
RESEARCH OBJECTIVES
surplus of car- since it
bon and limited can reduce environmentalfriendly
for vital nutrients op erat ion recovery process using
such as nitrogen, cost and chloroform, sodium
phosphorus and also reduce hydroxide and enzyme
sulphur. Under the solvent
these conditions damage To characterize the
cells cannot grow to health recovered PHA
but they do accu- and envi-
mulate carbon- ronment. To develop mass and
based polyesters. Moreo- energy balance for the
PHA production ver to get recovery process
by using Coma- a higher
monas sp. will be purity, a
conducted using purification step could gated using high pres-
mixed organic be added to the process. sure chromatography
acids obtained In this study, different (HPLC), gas chromatog-
P olyhydroxyal-
kanoates (PHA) is a
complete biodegradable,
by anaerobically treated
palm oil mill effluent
(POME) which replaced
initial conditions, wash- raphy (GC), differential
ing step, optimization scanning calorimetry
on recovery, optimiza- (DSC), thermogravi-
biocompatible, micro- the costly conventional tion on g force and de- metric analysis (TGA),
bial thermoplastic which carbon substrate. The velop the energy mass molecular weight
has potential to replace objectives of this study balance will be done (GPC), scanning elec-
petroleum-derived ther- are to develop an alter- using Design Expert tron microscopy (SEM)
moplastics. PHA is an native process recovery software. The proper- and transmission
excellent plastic option; of intracellular PHA, to ties of PHAs are highly electron micros-
a clean energy alterna- simulate and model the dependent upon their copy (TEM). ✳
tive with no emissions optimise recovery tech- recovery techniques;
of greenhouse gases, niques and to develop hence, biodegradable
which helps in address- mass and energy balanc- polymer having a wide
PICTURE
[SUPERVISOR] Professor Dr Mohd Ali Hassan ▶ Sodium hydroxide extraction

Elmy Nahida Binti
b i o m at er i a l Othman
elnada2712@gmail.com
CONTROLLED DEGRADATION OF
POLYHYDROXYALKANOATES BY
STEAM HYDROLYSIS FOR
CHEMICAL RECYCLING To identify the potential of PHA
chemical recycling by steam
hydrolysis and to characterize the
RESEARCH OBJECTIVES
P olyhydroxyal-
kanoate (PHAs) has
ducible biopolymer able
to contribute very signif-
namely pyrolysis and
hydrolysis. Pyrolysis and
hydrolysis products
unique characteristics icantly in the biomedical enzymatic hydrolysis of To investigate the effects of high
of thermoplastic, biode- applications especially PHAs have been exten- pressure and super heated steam
gradable and biocom- in tissue engineering sively studied; however
hydrolysis parameters on the molecular
patible biopolymer that (Chen and Wu, 2005). It steam hydrolysis of PHA
can be produced intra- is a good alternative to is yet to be studied. Deg-
weight of PHB and its characteristics
cellularly by microor- plastic petroleum which radation of PHA in this
ganism and some plant is non-biodegradable study, involved with the To identify the degradation behavior
species. This promis- and currently deplet- concept of the material of PHA steam hydrolysis and to
ing biopolymer has ing. This biodegrad- conversion to molecules propose the mechanisms involved in
been researched since able carbon reserve that built up the origi- steam hydrolysis of PHA
its discovery in 1920s, plays important role in nal material or lowering
remarkably starts the the environmental car- of its origin molecular
To prepare and characterize a
green revolution of non- bon storage. Single use weight (Ariffin et al.,
petrochemical aliphatic of bioplastics does not 2010) and (Ebdon and
product produced from recycled
polyester that is produc- support the sustain- Eastmond, 1995). This PHAs for slow release mechanism
ible via fermentation bi- ability of the carbon cy- research is aimed at re-
osynthesis and has been cle, therefore chemical covering low-molecular
commercialized as early recycling of bioplastic weight polymers with
as in 1962. Dawes (1986) is proposed. Chemical hydroxyl and carboxyl experiments will be constant and activation
in his monograph em- recycling is a process to chain-ends from polyhy- conducted using auto- energy based on relative
phasized that this com- depolymerize polymers droxyalkanoates (PHAs) clave as the reactor. The molecular weight of pol-
pound acts as a reserve to low-molecular-weight by high-pressure steam hydrolysis products will ystyrene as the standard.
material and intermo- materials before repo- (HPS) hydrolysis. These be characterized and the The hydrolyzates will be
lecular molecule that lymerizing it into new low molecular weight effects of the parameters characterized by using
is highly reduced and polymers. By chemi- polymers can be used as toward the target prod- 1
H and 13C NMR, FTIR
able to exert negligible cal recycling, cascade feedstock for repolymer- uct formation will be and DSC. Mass balance
osmotic pressure. The utilization of polymers ization and other appli- investigated in details. for the PHAs hydrolysis
constituent of PHAs, could be introduced cations. Degradation of The analysis of molecu- will also be studied. At
polyhydroxybutyrate before they are finally PHAs by HPS hydrolysis lar weight will be carried the end of this study, it
acid (PHB) is biocom- being released to the will be controlled by sev- out using gel permeation is expected that the deg-
patible with human as it environment. Several eral parameters, namely; chromatography (GPC) radation mechanisms
is also a built compound methods have been used temperature, pressure and the degradation is for PHAs hydrolysis can
of blood, made this pro- to depolymerize PHAs, and retention time. The projected theoretically be proposed, with the
to follow autocatalytic selective formation of
BACKGROUND PICTURE hydrolysis mechanism targeted products
[SUPERVISOR] Dr Hidayah Ariffin with the identification for chemical re-
▶ Prepared hydrolyzate samples in
NMR tubes for proton NMR analysis of critical point, rate cycling. ✳

Che Mohd Hakiman
Bin Che Maail
cm.hakiman.cm@gmail.com
ENHANCEMENT OF FERMENTABLE SUGARS

RECOVERY FROM OIL PALM FROND PETIOLE
O il palm frond
(OPF) is one of
the major crop residues
age effect, prior to juice
extraction will also be
studied in this case. En-
skinning of OPF may the storage period for
aid to increase the juice less than 48 hours and
release from the frond were sampled every 6
it hard to press besides
lesser moisture will be
retained. The longer of
which are produced an- zymes are introduced compared to without hours, the free sugars period storage will also
nually in Malaysia. Cur- to alter the structure of de-skin treat- leave the OPF
rent strategies of man- OPF petiole for better ment. The prone to micro-
aging the residues are extraction. In this study, skin of fronds bial attack which
RESEARCH OBJECTIVES
by formulating it into two enzymes are applied actually is re- To pre-treat the OPF leads to dra-
ruminant feed. Years which are cellulase and ally tough and petiole prior to juice matic decrease
back, the residues simply pectinase. The use of the presence of sugar content.
extraction in order to
discarded and burnt due the enzymes is expected might limit Free sugars from
to the cost burden. Later, to release more sugar the moisture
improve recovery yield OPF are promis-
ongoing research found yield better besides to escape when ing prospects for
that the OPF juice con- get a clearer juice. Ther- pressing is To study the effect the development
tains high fermentation mal treatment is done done. De-skin of fermentation
of OPF petiole
sugars such as glucose, to liberate remaining also aid in the industry. The re-
storage period on
sucrose and fructose sugars from the pressed convenience liance on this al-
that can be used for an- fibre. Pressed fibre will of pressing juice recovery ternative source
other rising industries. be soaked in 70 °C hot system since of raw material
Therefore, this research water and pressed again the hard part not only will be
study is proposed to after the first press. has been re- economical but
optimize the sugars re- The mechanical pre- moved. Storage of OPF will be analyzed and contribute to the ef-
covery during juice ex- treatment proposed to prior to extraction may determined to see the ficient management of
traction with the means remove the skin of OPF regulate more free sug- best peak of free sugars agricultural residues in
of enzymatic, mechani- as the presence of the ars besides add on effect generated. Any period the country. Usage of
cal pre-treatments and skin cause difficulties in on the moisture content longer than 48 hours will OPF as sugar alterna-
thermal treatment. Stor- pressing the OPF. De- inside the OPF. Giving wither the frond making tive to the fermentation
definitely would not
[SUPERVISOR] Dr Hidayah Ariffin conflict to the food con-
sumption if rela-
tively compared
to the sugar from
sugar cane and
any other sugar
producing food
plant. This study
is hoped to con-
tribute to a better
OPF sugar recov-
Oil Palm Frond
▶
ery with through

◀ Oil Palm Pressing Machine, ▶ the convenient
Frond Petiole OPF Juice and
OPF Fibre
steps pro-
posed. ✳

Mohd Nor Faiz b.
b i o m at er i a l Norrrahim
Faiznorrrahim@gmail.com
IMPROVED MECHANICAL AND

THERMAL PROPERTIES OF
POLYHYDROXYALKANOTES
BY BLENDNG WITH OTHER
POLYMERS cibility of the polymer
blends will be deter-
mined morphologically
exhibit better thermal
and mechanical prop-
erties compared to the
by using scanning elec- homopolymer of PHA.
tron microscopy (SEM). It is also expected, that
mal properties of this bi- Besides that, thermal these polymer blends
opolymer. In this study, properties of the poly- can be separated by py-
polymers such as poly
(methyl methacrylate)
S
everal excellent re-
search done by the
(PMMA), polyethylene
(PE), and natural fiber To improve the thermal and mechanical
RESEARCH OBJECTIVES
scientist on the bioplas- is used to enhance the properties of PHA by blending with
tics has been considered mechanical and ther- PMMA, PE and mesocarp fiber and to
as the solution for the mal properties of the propose the applications of PHA blends
serious environmental blends. Mesocarp fiber based on their properties
> Hydraulic Hot problem. Good strength, is selected as the natu-
Press Machine
low production cost, and ral fiber source. To the To determine the effect of
good resistant towards best of our knowledge,
compatibilizer on the thermal and
chemical and biological there is no research done
attack are the impor- so far on the blending
mechanical properties of PHA blends
tant things need to be of PHA with mesocarp
To study on the thermal
considered to produce fiber for the production
bioplastics. Poly-hy- of biocomposite. Several
degradability of the PHA blends
droxyalkanoate (PHA) is polymeric blend ratios
a biodegradable polymer (0-50%) will be prepared mer blends will be tested rolysis after its use. In
with some advantages of using Thermo Haake by thermogravimetry order to study the sepa-
> Instron excellent biodegradabil- mixer and hydraulic hot (TG), differential scan- ration, thermal degra-
Mechanical
ity with high crystalline press machine. The ef- ning calorimetry (DSC) dation of these polymer
Analyzer
and good thermal prop- fects of plasticizer and and thermo mechani- blends will be conducted
erties, however it is not compatibilizer incorpo- cal analysis (TMA). by using glass tube oven
practical to be applied as ration in the polymer Mechanical properties and the pyrolyzates and
commodity plastics due blends will be studied. such as, tensile strength, result will be analysed
to its brittleness and nar- Maleic anhydride and si- Young’s modulus and by Nuclear Magnetic
row processing window. lane coupling agent will elongation at break of Resonance (NMR). It is
Blending of PHA with be used in this research the polymer blends will hoped that the applica-
> Thermo other polymers may en- as the compatibilizers be determined by us- tion of bioplastics and
Haake Mixer hance the mechanical while poly ethylene gly- ing Instron Mechanical biocomposite can be
and col (PEG) will be used analyzer. The polymer extended for com-
[SUPERVISOR] Dr Hidayah Ariffin ther- as plasticizer. The mis- blends are expected to modity use. ✳

Noor Ida Amalina
Ahamad Nordin
idamalina@yahoo.com
UTILIZATION OF OIL PALM

MESOCARP FIBER FOR which is biocomposites.
Mesocarp is the fibrous
mass left behind after
oils extraction. Current-
BIOCOMPOSITE
ly, mesocarp is burned as
fuel to generate energy
lion tonnes of mesocarp for mills. However, this
fiber, 4.3 million tonnes practice creates another
palm kernel shell and environmental problem
53.1 million tonnes of due to incineration and
palm oil mill effluent are emitted gas. Biocom-
generated by oil palm in- posites are defined as
M alaysia is blessed
with fertile soil
and abundant natural
dustries.
Oil palm biomass ap-
pears to be a very prom-
the materials made by
combining natural fiber
and petroleum derived
resources. Malaysia also ising alternative as a non-biodegradable pol-
is one of major producer source of raw materials ymer or biodegradable
To clarify effects of high pressure in palm oil industries. in Malaysia. In recent polymer. Natural fiber
RESEARCH OBJECTIVES
and super heated steam treatments According to USDA years, there has been an as filler in polymeric
on the characteristics of mesocarp (2007), Malaysia and In- expanding search for matrix offers several
fibers donesia are major con- new materials with high advantages such as high
tributor which is about performance at afford- bending strength, good
To prepare and characterize 87% of world palm oil able cost and increased thermal and insulation
biocomposites derived from production. Despite of attention has been paid properties, lower den-
steam treated mesocarp fiber and the economic contribu- to the use of natural sity, non-toxic, ease of
tion, palm oil industries polymers also lignocel- recycling, flexible usage
polypropylene
generate huge amount lulosic fibers. and lower cost. Meso-
Depolymerization of biocomposite of solid waste and efflu- This study will focused carp component con-
via thermal degradation method ent. In 2006, about 17.4 on oil palm mesocarp sists of 31.9% cellulose,
million tonnes empty fiber to being utilize as 35.7% hemicellulose and
fruit bunch, 10.7 mil- value added product 32.4% lignin which >

> Hot pressed > Injection moulding
samples for tensile test samples for bending test
PICTURE (TOP)
▶ Biocomposites samples prepared for tensile and bending test
PICTURE (BACKGROUND)
▶ Abundant of mesocarp available in palm oil mill
make it a potential ma- of the composite. By low moisture content morphology proper-
terial for biocomposite. solving the mentioned which makes grinding ties of biocomposite
Only the drawbacks of drawback, good thermal and mixing processes will be studied. In this
natural filler are weak and mechanical proper- easier. By removing study, polypropylene
interaction between fill- ties of the biocomposite moisture content, it is (PP) is used to mix with
er and polymer matrix can be produced. expected that hydroxyls untreated and steam
and filler dispersion due Steam treatment will group is removed and treated mesocarp fiber.
to different properties of be used to improve the promote better char- Finally, the characteris-
natural fibers and poly- characteristic of the acteristic of mesocarp tic of the biocomposites
mer. Natural fiber holds fiber in term of stabil- fibers. The treated sam- will be analysed using
hydrophilic character- ity, composition, and ple will be grinded and TGA, DSC, tensile and
istic due to presence of provide a stable bond sieve to desire particle flexural strength and
hydroxyl group resulting between the fiber and size. Effect of fiber size, morphology by SEM. ✳
poor interfacial adhe- the polymer matrix. fiber homogeneity, fiber
sion with hydrophobic It is expected that the ratio towards ther-
polymer matrix. In the treatment can enhance mal, mechani-
same time also, aggrega- the interfacial bonding cal and
tion of fiber easily can strength of the compos-
occur. Natural fibers ite. Two approaches will
also have high capabil- be conducted which are
ity of water sorption due high pressure steam
to cellulose, hemicel- and super heated
lulose and lignin com- steam. The opti-
ponents. The hydroxyls mum treatment
group which presence time, tempera-
in the fibers components ture and pres-
leading to formation of sure will be
hydrogen bonds with determined.
water. This may lead Steam treated
to poor physical and mesocarp fib-
mechanical properties ers have very
▶ Cellulose mesocarp fiber

BIOFERTILIZER
RESEARCH GROUP
 RESEARCH SYNOPSIS  PRINCIPAL RESEARCHER
The Biofertilizer Research Group is leading in advanced

composting process and waste recycling technology for
agriculture purposes and wastewater treatment. The
group main focus is on effective organic wastes recycling
technology to convert organic waste into organic fertilizer.
Currently, the researches on oil palm biomass composting
and pyrolisis process are conducted in the group under
the collaboration between UPM and KIT. The economic
feasibility analysis also been conducted for both researches PROFESSOR DR Dr Azhari Samsu
to promote zero discharge in oil palm industry. ✳ MOHD ALI HASSAN Baharuddin

BIOFERTILIZER
biocompost biochar
Large Scale small Scale
Mohd Mohd Ahmad Mohd

Kamahl Najib Amiruddin Ridzuan
production economic aspect :

of enzymes zero discharge
Mohd Nor Yoshizaki

Huzairi Hashimah Tatsuya

Mohd Huzairi
b i o f ert i l i z er Mohd Zainudin
huzzairie@yahoo.com
ISOLATION AND CHARACTERIZATION

OF CELLULASE AND XYLANASE
FROM OIL PALM EMPTY To investigate the microbial
RESEARCH OBJECTIVES
community during the oil
palm empty fruit bunch
FRUIT BUNCH COMPOST compost process
To isolate and characterize the

expressed cellulase and xylanase
from isolated microbes
M alaysia has be-
come the largest
oil palm producer with
ducing cellulase and xy-
lanase was investigated.
In this study, DNA was
from EFB compost. The
DNA is subsequently
cloned into vector (fos-
To screen and identify cellulase
the production of about extracted and purified mid) to construct the
and xylanase gene through
18 million tonnes per from EFB compost by metagenomic libarary. metagenomic approach
year and about 47% of DNA soil extraction kit. From the metagenomic
world’s supply. Besides The isolated DNA was library, the genes en-
producing oil, it has used for determining coding cellulase and
generates abundant of the microbial popula- xylanase were screened Hopefully in the future,
waste such as Palm oil tion through microbiota identified by polymer- these finding will pro- ▶ Metagenomic library of
mill effluent (POME), analysis by using cul- ase chain reaction or vide the good enzyme to EFB compost for cellulase
Empty fruit bunch ture-independent of 16s activity screening. The improve the feasibility of
(EFB), Mesocarp fiber rRNA gene amplified di- findings of this study lignocellulose biomass
and Palm kernel shell. rectly from the compost. helped to understand conversion. ✳
EFB is one the largest 1500-bp 16S rRNA PCR the microbial popula-
waste produce in the products were cloned tion throughout the
mill. Previously, EFB has and sequenced. composting process and
been dumped for soil Subsequently, with re- identify the microbes
mulching in the planta- gard to the results ob- that can produce
tion area. One way to tained from microbiota cellulase and xy-
create value added prod- analysis, screening and lanase. It is also
uct from these waste are isolation of bacteria helped to find
through the compositing producing cellulase the ligno-
using EFB with anaero- and xylanase was done. cellulosic
bic sludge POME. EFB Several microbes that enzyme
compost is manageable are able to express cel- by isolat-
product which can be lulase and xylanase have ing genes
use as soil amendment been isolated. In order encoding
and organic fertilizer. In to identify potential cellu-
composting, lignocel- cellulase and xylanase lase and
lulose material breaks enzyme from noncultur- xylanase
down due to the exist- able microbes, metagen- which was
ence of aerobic thermo- omic of EFB compost screened
philic bacteria. The mi- was done. Metagenome f r o m
crobial population and involves the extraction metagem-
microbes capable of pro- of metagenomic DNA onic library.

Ahmad Amiruddin
Mohd Ali
b i o f e rt i li z e r
amiruddin_99@yahoo.com
ENHANCED BIOMETHANE PRODUCTION

THROUGH OIL PALM BIOMASS UTILIZATION
T he palm oil industry
represents the larg-
ing diesel as in the cur-
rent practice. This will
better utilization of bio-
mass resources can be
To improve methane production
and yield from palm oil mill effluent
RESEARCH OBJECTIVES
est agro-economic sec- free up high calorific achieved in the mill. In (POME) anaerobic batch and
tor in Malaysia. Howev- value biomass such as terms of potential, mills continuous fermentation systems by
er as the demand for oil shells which are typically could be self-sufficient
utilizing added oil palm biomass
palm products increase used as fuel source to in terms of energy, capa-
globally, more atten- be sold to other parties. ble of providing surplus To evaluate performance of biogas
tion is being paid to the In addition, any surplus electricity to the grid, as scrubbing system using mill treated
impact of huge wastes electricity may be con- well as having unused river water and biochar produced
generated from the in- nected to the grid for and high value biomass from low-cost technology
dustry such as palm oil external use and gener- such as shells available
mill effluent (POME), ate additional income to for sale. All this trans- To produce biochar and activated
oil palm empty fruit the mill and industry in lates into extra revenue carbon from oil palm empty
bunch (EFB), mesocarp general. while simultaneously fruit bunch (EFB) using low-cost
fiber and oil palm fronds The first and second enhancing the image of technology and excess steam
(OPF) towards the envi- objectives of the study the palm oil industry
ronment. In the conven- focus on the method through more environ-
tional system, POME is for improving methane mentally sustain-
treated using expansive production and yield in able practices. ✳ [SUPERVISOR] Professor Dr Yoshihito Shirai
open lagoons and/or both batch and continu-
tanks prior to discharge ous anaerobic fer-
to the river or water- mentation systems > SEM image of oil palm biomass after fermentation
course. The solid wastes of POME through
on the other hand are utilization of added
being utilized as boiler oil palm biomass.
fuel, mulching agent The third objective
in the oil palm planta- involves the study
tions or incinerated. and identification
Hence this study aims of microbes and
to utilize these oil palm enzymes involved
biomass or residues spe- in the fermenta-
cifically POME and EFB, tion using Scanning
which are the largest Electron Micro-
by-products of the palm scope (SEM), De-
oil extraction process naturing Gradient
to enhance biomethane Gel Electrophoresis
production. (DGGE), Fluores-
Biomethane can be con- cent In-Situ Hybridi-
verted into electricity via zation (FISH) and
gas engines and utilized microbiota analyses.
by the mill as its source From the research,
of energy rather than us- it is envisaged that

Mohd Ridzuan
b i o f ert i l i z er Othman
dh_zuan@yahoo.com
APPROPRIATE TREATMENT OF PALM OIL MILL

FINAL DISCHARGE WASTEWATER AS
RECYCLED WATER FOR THE MILL
TO ACHIEVE ZERO DISCHARGE
I n palm oil industry,

huge amount of wa-
as appropriate treatment
of palm oil mill final dis-
processes that encour-
age the removal of COD, To study an effective dosing of
RESEARCH OBJECTIVES
ter have been utilized charge wastewater have color, suspended col- organic and inorganic coagulant
for palm oil sterilization been evaluated in order loids and organic matter usage for the treatment of palm oil
and extraction process. to recycled water for in final discharge waste-
mill final discharge wastewater to
The processing system the mill to replace fresh water which can lead to
achieve zero discharge in the mill
has been applying widely river water. Current turbidity, odor and taste
in Malaysia for year. chemical treatment used problems. In this re-
It has been estimated at the mill will be used search Jar Test is used to To develop low cost environmental
that around one ton of to treat final discharge to determine the optimum friendly methods by using organic
fresh water was needed achieve zero discharge. operating conditions for coagulant to treat palm oil mill
for processing every Activated carbon is used final discharge waste- final discharge wastewater
ton of fresh fruit bunch as absorbent material water by optimizing
(FFB). As a return, huge due to its large number value of pH, dosage of
amount of wastewa- of cavernous pores that coagulant and activated
ter has been generated, provide a large surface carbon used and mixing
treated and discharged area relative to the size time to improve the per- ▶ Zeolite for final disharge
wastewater treatment
to the river every day. of the actual carbon formance and/or capac-
Current treatment sys- particle and its visible ity of existing treatment
tem applying in oil palm exterior surface. A Jar systems and to reduce
industry is using river Test Method is used to capital expenditure
water to use for mill. In stimulate the coagula- on new treatment
present study, the effect tion and flocculation systems. ✳
of coagulant and acti-
vated carbon application [SUPERVISOR] Professor Dr Mohd Ali Hassan

Tatsuya Yoshizaki
Yoshizaki-tatsuya@edu.life.
kyutech.ac.jp
ZERO DISCHARGE ON
PALM OIL INDUSTRY
To evaluate economic performance of
RESEARCH OBJECTIVES
integrated technology of biogas and
P alm oil industry is

known as a signifi-
cant agricultural indus-
compost production in palm oil mill
try in terms of economic

To produce biochar and activated
benefit for Malaysia. carbon from oil palm empty
However, on the other fruit bunch (EFB) using low-cost
hands, palm oil mills technology and excess steam
have created environ- Its common practice is
mental problems due to to disposal into planta-
huge quantities of pol- tion as nutrients recycle,
To evaluate economic viability of
luted waste materials. however it leads to pol- integrated zerodischarge system in
Palm oil mill effluent lution problems such as palm oil mill
(POME) is highly indus- eutrophication and an
trial waste water with increase of toxicity in
several technologies
high organic content. It the soil. Although over
including biogas from
has been recognized as the past decade, the
POME as renewable en-
a main contributor of palm oil industry has
ergy, composting EFB
green house gas (GHG) developed proper utili-
through mixing with its byproducts actively.
emission especially con- zations of these byprod-
POME anaerobic sludge, Furthermore, this dem-
sisted of methane from ucts, it is less research
and carbonizing EFB. onstration can improve
open pond and tank to ensure the economic
These technologies surround environment
treatment system. Emp- viability by introduc-
should be integrated by avoiding improper Biodiversity and Eco-
ty fruit bunch (EFB) is ing these technologies.
and support each other. treatment of POME systems Conservation
a residue after milling The idea of zerodis-
This integrated system and inefficient disposal (BBEC) Program with
fresh fruit bunch (FFB). charge is to introduce
can provide a good so- of EFB. This study is Japan International
lution for palm applied in Sandakann, Company Agen-
[SUPERVISOR] Professor Dr Yoshihito Shirai oil mill to utilize Sabah, under Bornean cy (JICA). ✳

Mohd Najib Ahmad
b i o f ert i l i z er Master Student, Semester 6
mnajib@mpob.gov.my
Composting of Oil Palm Frond

by enhancement of POME
anaerobic sludge
A total of 54.44 mil-

lion tons of oil
tion and generation of
waste in oil palm sector
function of the accumu-
lation of heat generated
tures via an investigation
of bacterial species di-
RESEARCH OBJECTIVES
To study the microbial
palm fronds had been and to increase palm oil metabolically and deter- versity at different com-
population during
generated from the palm productivity. In com- minant of metabolic ac- posting temperatures by
oil industry in 2008. Re- posting, providing a tivity. The compost was using denature gradi-
production of palm
alizing the potential and stable product that is entering a thermophilic ent gel electrophoresis biomass biofertilizer
abundance of fronds as high in nutrients which phase, with temperature (DGGE). The result of
sources of renewable are easily accessible by recorded at 52 °C after 6 DGGE analysis revealed To develop rapid
raw materials, research plants is essential. The days of composting. The that the main microbes open system for oil
to produce Biocompost basic process control thermophilic condition during co-composting palm fronds (OPF)
from oil palm frond objective is to maximize encourages the com- of chipped frond and composting.
(OPF) had been initi- microbial activity at the posting process with the POME anaerobic sludge
ated. Co-composting expense of the waste carbon to nitrogen ratio belong to group of
of palm biomass into being treated. This is decrease from 80 to 15.3 Gammaproteobacteria. ✳
microbial based biofer- equivalent to maximize during 60 days of com-
tilizer is essential to metabolic heat output. posting. Process stability
reduce the impact of In the self-heating eco- is favored by moderate
environmental pollu- system, temperature is a thermophilic tempera-
BACKGROUND PICTURE
[SUPERVISOR] Professor Dr Mohd Ali Hassan ▶ Oil palm frond compost

Nor Hashimah
Abdul Rahman
hashimahrahman@gmail.com
BIODEGRADATION OF LIGNIN BY ISOLATED

LIGNINOLYTIC BACTERIAL STRAIN FROM
OIL PALM PLANTATION SOILS
L igninocellulose are breakdown is thought depolymerize the high To screen, isolate and
RESEARCH OBJECTIVES
mainly consists of to occur by concomitant molecular weight back- characterize ligninolytic
lignin, hemicellulose action of ligninolytic en- bone of the lignin poly- bacteria strains from oil palm
and cellulose (Betts zymes. mer because the bacte-
plantation soils
et. al., 1991; Sun and Microorganisms that rial cells do not secrete
Cheng, 2002). Lignin is degrade lignin through lignin-depolymerizing To study the effects of
well-known for resist- an oxidative process are enzymes unlike fungi
different temperature, pH
ance to microbial deg- fungi, actinomycetes which secrete extracel-
radation because of its and to a lesser extent, lular enzymes called
and substrates on ligninolytic
high molecular weight bacteria. In the literature ligninases (Vicuna, enzymes production from
and presence of vari- review, white rot fungi 1988). Still, bacterial isolated bacteria in submerged
ous biologically stable have received extensive lignin degradation sys- fermentation
carbon-to-carbon and attention in research for tems have ligninolytic
ether linkages. Gener- ligninolytic enzymes be- potential because it con-
ally, lignin contains cause of their powerful sists of many unique and
three aromatic alcohols lignin-degdrading enzy- specific enzymes with
which are coniferyl matic systems (Hatakka, the ability to catalyze tial ligninolytic bacteria standing of bacte-
alcohol, sinaply and 1994). Even so, fungi the production of vari- strains. The observation rial communities in the
p-coumaryl and there are unstable in practi- ous useful compounds. that several soils bac- samples will be investi-
are many problems and cal treatment under Bacteria are important teria with the ability to gated by using culture-
difficulty in dissolving extreme environmental to be studied for lignino- degrade aromatic com- based techniques of mi-
lignin without destroy- and substrate conditions lytic potential because of pounds are also able to crobiota analysis. Based
ing it and some of it such as, oxygen limita- their immense environ- degrade lignin provides on the lists of the total
subunits because of its tion, high extractive, mental adaptability and a possible link between microbial community in
exact chemical structure higher pH and lignin biochemical versatility aromatic degradation the environmental sam-
is difficult to ascertain. concentration (Nagar- (Lisboa et. al., 2005). and lignin degradation ples, isolation of poten-
Lignin iss the most re- athnamma et. al., 1999). There are several types of (Timothy et. al., 2010). tial ligninolytic bacteria
calcitrant to degrade Hence, in some studies, oil palm plantation soils In order to explore the strains can be conduct
because of its highly or- it shows evidence about in Felda Serting Hilir, full potential of ligni- and classified according
dered crystalline struc- the bacterial strains can Negeri Sembilan were nolytic bacteria strains to phylogenetic analysis
ture is more resistant to degrade the low mo- chosen as samples in contained in oil palm (Fleske et. al., 1998;
hydrolysis than hemi- lecular weight portion of this project for screening plantation soils, a clear Fritsche et. al.,
cellulose. Thus, lignin lignin, but are unable to and isolation of poten- and complete under- 1999). ✳
[SUPERVISOR] Dr Nor ‘Aini Abdul Rahman

BACKGROUND PICTURE ▶ Pure colony of ligninolytic bacteria

Mohd Kamahl
b i o f ert i l i z er Mohamad Kamarudin
kamahl.mk@felda.net.my
Improvement in Co-Composting
Process of Pressed-Shredded EFB
and Raw POME from Continuous
Sterilizer System
To investigate the effect
RESEARCH OBJECTIVES
of various OPF bulking
T he continuous steri-
lizer system is a
EFB with the addition
of raw POME was an
ing continuous steriliz-
ing empty fruit bunch
size in composting of
POME under different FFB
high performance sys- option to solves wastes (EFB). In order to get sterilization process
tem in palm oil extrac- accumulation problems better understanding
tion. In future trend, in the mills. Currently on physicochemical in To investigate the feasibility
more and more mills composting technology continuous sterilizing
of biocompost production
in Malaysia will install- on empty fruit bunch empty fruit bunch, de-
ing such fresh fruit (EFB) and raw POME tail study on chemical
from co-composting of oil
bunch (FFB) processing still in the infrant stag- and structural proper- palm EFB with raw POME
system. However, The es. The decomposition ties has been conducted.
non-ponding system in of empty fruit bunch The scanning electron
continuous sterilization (EFB) in acidic condi- microscopy (SEM) and
system has generated tion may inhibit micro- transmission electron play an important role process. This study was
huge amount of empty bial decomposition rate. microscopy (TEM) give in effective composting targeting to deliver a
fruit bunch (EFB) and Therefore, further study a full picture on struc- since raw POME lack of good composting pro-
raw POME that had cre- regarding microbial de- tural disruption under microbes. Hence, effec- cess for raw POME and
ate problem to the mills. composition on empty sterilization. For com- tively utilizing microbial empty fruit bunch
Therefore, composting fruit bunch (EFB) was posting process, micro- seeding was a key for (EFB) for industri-
of continuous sterilizer important for utilizat- bial seeding method successful composting al application. ✳

2011 IN PICTURES
▶ Prof Dr Mohd Ali, Prof. Dr Suraini and Dr Hidayah ▶ Asian Congress on Biotechnology, Shanghai, China
during visit to Shanghai Tauto Biotech Company (10th – 16th May 2011)
▶ Prof. Suraini with Y.Bhg. Dato’ Dr. Rosli Mohamad (Secretary General,
Ministry of Education), UNESCO Representative and YAB Tan Sri Dato’
Hj. Muhyiddin Hj Mohd Yassin (Deputy Prime Minister of Malaysia. ▶ Prof Dr Suraini and Ms. Dayang Salwani at 10th International
High-Tea after the Hari UNESCO Malaysia 2011 on 18th November 2011. Sago Symposium, Bogor, Indonesia (October 2011)
▶ IEA Hydrogen Implementing Agreement (HIA), Task 21 Meeting, Singapore ▶ High-Tea after the Hari UNESCO Malaysia 2011
(28th February – 1st March, 2011) on 18th November 2011

2011 IN PICTURES
▶ EB Group members at International Invention, Innovation ▶ Mitra and Yee at International Invention, Innovation
& Technology Exhibition, ITEX 2011 & Technology Exhibition, ITEX 2011
▶ Syahinaz and Shima at Pameran Reka Cipta ▶ EB Group members at Seminar New Generation and Breakthrough
dan Penyelidikan (PRPI) 2011, UPM Serdang Technology in GC, GCMS, GCMS-MS & ICPMS 1
▶ Memorial picture with other participants at Shell Intervarsity Student ▶ Shima at Malaysia Technology Expo 2012
Paper Presentation Contest (S-SPEC 2011) at UTM Skudai

Find out more about EB Group online…
www.ebgroup.upm.edu.my
G
EDITORIAL TEAM
EB GROUP
CHIEF editor Prof. Ali Hassan

editorial TEAM Dr Hidayah, Faiz, Azwan, Amelia, Aen
special thanks Mdm. Rosema, Shima, Ridzuan
DESIGN & layout EB Group Editorial Team

EB GROUP
www.ebgroup.upm.edu.my
ENVIRONMENTAL BIOTECHNOLOGY RESEARCH GROUP

Faculty of Biotechnology and Biomolecular Sciences
Universiti Putra Malaysia 43400 Serdang, Selangor MALAYSIA
Phone : +603-8946 7590 Fax : +603-8946 7593

Research Report 2011: Environmental Biotechnology Research Group

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ENVIRONMENTAL BIOTECHNOLOGY RESEARCH GROUP

RESEARCH REPORT 2011

Message from the

group Big Picture

▶ Bioplastic Pilot Plant Holding Tank

God bless. Wassalam.

Professor Dr Mohd Ali Hassan

RESEARCH REPORT 2011 > EB GROUP 01

02 RESEARCH REPORT 2011 > EB GROUP

c a r b onis a tion bio- f u el

bio- compo sit e

c e ll ulos e biosugar biohydr o gen

MARDI / UPM TPU FARM SCHEDULED GW COLLECTION

SCHEDULED & VOLUNTARY USED COOKING OIL

residential college food waste

UPM : universiti Putra Malaysia

 TO MARKET MARDI : Malaysian Agricultural Research

04 RESEARCH REPORT 2011 > EB GROUP

PARASITIC USE - upm TPU / MARDI FARM / MPSJ LANSCAPING ( BIOCOMPOST )

PARASITIC USE - BIOREFINERY PLANT

SOLID & LIQUID DIGESTATE

CTRICITY GENERATED FROM

NON-RECOVERABLE MATERIAL / Contraries

RESEARCH REPORT 2011 > EB GROUP 05

Mior Ahmad Mohd

ORGANISATION prof. ali

EB GROUP KEY // GROUP LEADER

06 RESEARCH REPORT 2011 > EB GROUP PRINCIPAL RESEARCHER

Nur Ain Dr Nor'Aini

phd PH.D. STUDENT ra RESEARCH ASSISTANT

msc MASTER STUDENT

Professor Dr Mohd Ali Hassan

08 RESEARCH REPORT 2011 > EB GROUP

Professor Dr Suraini Abd Aziz

RESEARCH REPORT 2011 > EB GROUP 09

Dr Phang Lai Yee

contacts 2011 patent

10 RESEARCH REPORT 2011 > EB GROUP

contacts 2011 Seminar presented

RESEARCH REPORT 2011 > EB GROUP 11

Dr Nor’Aini Abdul Rahman

2011 Seminar presented (poster)

12 RESEARCH REPORT 2011 > EB GROUP

Dr Azhari Samsu Baharuddin

academic qualification 2011 consultancy

RESEARCH REPORT 2011 > EB GROUP 13

YOSHIHITO SHIRAI, ph.d. Hiroaki Ogawa Haruo NISHIDA, Dr. Eng

Umi Kalsom Toshinari Helmi Wasoh @

Faculty of Biotechnology Department of Biological Functions Faculty of Biotechnology and Biomolecular

14 RESEARCH REPORT 2011 > EB GROUP

Tel: +603-9051 4421 Tel: +81-93-695-6238 Tel: +81-93-695-6066

Rosema Ghazali Prof. Shin WATANABE Dr S. Karuppuchamy

GROUP SECRETARY FELDA BIOMASS POSTDOCTORAL RESEARCHER

Tel: +603-8947 1182 Tel: +603-2162 3838 Tel: +603-8946 7593

RESEARCH REPORT 2011 > EB GROUP 15

Current Position: Current Position:

Isnazunita Nazlina Haiza LIM SIONG

Current Position: Current Position: Current Position:

Nurul Kartini Farah Nadia Halimatun

Current Position: Current Position: Current Position:

16 RESEARCH REPORT 2011 > EB GROUP

PARTICIPANTS PROGRAM RESEARCH THEME HOST/LOCATION DURATION SPONSOR

Development of novel Assoc. Professor Kyushu Institute