Glucose Production of Cellulose Hydrolysis Enzymatically on Rice Straw

Novia, Hermansyah dan Selvia Aprilyanti

Environmental Technologies of Magister Chemical Engineering, University of Sriwijaya

Email : noviasumardi@yahoo.com, selviaaprilyanti@yahoo.co.id

ABSTRACT
Rice straw is a by-product of agricultural activities resulting in large enough quantities in
Indonesia. Rice straw has the potential to be used as raw material for alternative energy, namely
bioethanol or other alternative fuels. Rice straw contains cellulose and hemicellulose that can be
used to produce glucose by enzymatic hydrolysis. Enzymatic process is environmentally friendly
process renewable raw materials. Enzymatic hydrolysis performed using cellulase enzymes to
produce glucose. The purpose of this study was to determine the effect of hydrolysis time and
enzyme concentration used for glucose production from rice straw that had been pretreated by
alkali and ozonolisis. In this study, a 50 grams rice straw pretreated was dissolved in 100 ml of
medium and the pH was adjusted to 5. The parameters varied are the hydrolysis time 5,10, 15, 20
and 25 hours, whereas the concentration of enzyme added was 10%, 20%, 30% and 40% w / v of
the weight of rice straw. The results showed that the greatest glucose obtained was 0.08752 gr at
25 hours of hydrolysis and enzyme stater volume of 40% of the weight of rice straw.
Keywords : enzymatic hydrolysis, glucose, rice straw

I.

INTRODUCTION
Rice straw is one of the largest
agricultural wastes in southern Sumatra.
Rice production per hectare can reach 10-15
tons of dry matter each harvest, depending
on location and crop varieties. Each
kilogram of rice straw yields results in the
production of 1 - 1.5 kg of rice straw.
Global production of rice straw around 650975 million tons per year and is mostly
used as animal feed and as agricultural
waste (Binod et.al, 2009). Burning of rice
straw is one of the practical ways to address
waste rice straw, but this could increase air
pollution and negative impacts on public
health. Therefore, rice straw needs to be reprocessed into useful materials but does not
pose a problem for the environment.
Rice straw containing lignin, cellulose,
and hemicellulose. Cellulose content of rice
straw is quite large, which is about 35-45%,
so that rice straw can be used to produce
glucose (Galleti and Antonetti., 2011).
Presence of lignin in the straw will inhibit

the action of enzymes to convert cellulose

into glucose, therefore it is necessary to pretreatment (pre-treatment) to reduce the
amount of lignin in rice straw making
cellulose more easily react with the
enzyme. Conversion of cellulose to glucose
is done with the help of cellulase enzymes.
Cellulase enzymes will accelerate the
hydrolysis of cellulose to glucose (Fatma et
al, 2010).
Pretreatment technologies are grouped
into physical, chemical, physicochemical
and biological. In this study, conducted
physically Pre-treatment which aims to
minimize the size of the biomass and
reduce the crystallinity of cellulose, is done
by cutting and grinding. Pre-treatment is
then performed chemically by using a
solution of NaOH / alkaline (Yoswathana et
al, 2010; Roslam et al, 2011) and the
ozonolysys to degrade lignin and
hemicellulose in biomass (Cubero et al,
2009).

Hydrolysis is the process of solving the

polysaccharides in lignocellulosic biomass,
namely cellulose and hemicellulose into
sugar monomers constituent. Perfect
cellulose hydrolysis yield of glucose, while
hemicellulose monomer sugars produced
some pentose (C5) and hexose (C6).
Hydrolysis can be done chemically (acid) or
enzymatic. Enzymatic hydrolysis has
several advantages over acid hydrolysis,
among other things: no sugar degradation
hydrolysis results, a milder process
conditions (low temperature, pH neutral),
has the potential to provide high yields, and
equipment maintenance costs are relatively
low because no corrosive materials
(Taherzadeh and Karimi, 2007; Sun Y and
Cheng, 2002). Enzymatic process is
environmentally friendly process renewable
raw materials (renewable raw materials).
Currently, enzymatic hydrolysis is a very
promising technology to convert biomass
such as rice straw to glucose.
Aspergillus Niger is a fungi that produce
cellulase enzymes and can hydrolyze
cellulose. Aspergillus niger to produce
cellulase enzymes in rice straw media with
high enzyme activity (Ong et al, 2012).
This study aims to obtain the best operating
conditions on enzymatic hydrolysis of rice
straw to glucose by using Aspergillus Niger.
II. RESEARCH METHODOLOGY
2.1 Preparation of Raw Materials
Biomass in the form of rice straw
obtained from paddy fields in the
region MUSI 2 Palembang. Rice straw
was destroyed using grinding machine
(mill). Then dried in the hot sun for 10
days. Then heated in an oven at 45 C
to remove the water content in the
biomass. Once dry, the smaller the tool
grinder to a certain size and then sieved
to obtain the size of the straw 0.25 mm
(80 mesh).
2. 2 Alkaline pre-treatment
Rice straw that had been measured
lignin content and moisture content of
50 grams into the bottle lid (lid

erlenmeyer) and dissolved in 5%

NaOH solution of 500 ml (comparison
ratio (w / v) rice straw: NaOH = 1: 10).
Further samples were incubated in a
container in the water bath at a
temperature of 850C for 1 hour
(Yoswathana et al, 2010). After 1 hour,
samples were filtered and washed with
distilled water until neutral pH. Further
samples of solids dried in an oven at a
temperature of 1050C to constant
weight. lignin content of the final
sample is calculated using the Kappa
method.
2.3 Ozonolysys Pretreatment
Ozonation process is carried out at a
constant voltage, the 8500 Volt.
Treatment of oxygen flow rate and the
sample size of each variable at 5 L /
min and the sample size 0.177 mm 0.25 mm. Ozonolisis process rice straw
held in ozonation reactor. 50 gram
samples of rice straw in a state of
10% moisture content is inserted into
the
reactor
ozonolisis.
Before
mengozonolysis biomass, ozone levels
were analyzed first by iodometric
method. 2% KI solution into a tube
inserted into the analysis-1 (bottom
tube) and analysis of the 2nd tube (tube
top). Ozone gas flowed into the
ozonation reactor for 15 minutes. After
jetting 15 minutes, ozone is formed
analyzed from KI solution to the tube-1
with the iodometric method. While the
rest of the gas supplied to the 2nd tube
analysis. The content of residual ozone
ozonolisis reaction was analyzed as the
tube analysis to-1 with the iodometric
method. The samples were diozonasi,
analyzed the content ligninnya using
Kappa method.
2.4 Enzymatic hydrolysis
The results of pre-treatment put into
500 ml Erlenmeyer then added 100 ml
of distilled water and adjust pH 5. Then
heated in an autoclave at 100 C for 30
minutes. Rice straw pulp allowed to

cool. Cellulase enzyme is added

according to treatment variables into
mush the rice straw and Erlenmeyer
sealed with a cork. Treatment of
hydrolysis time, each consisting of the
5, 10,15, 20 and 25 hours, while the
treatment of the enzyme concentration
is 10%, 20%, 30% and 40%. Enzyme
concentration of 10% means that the
addition of the enzyme with 10% of
Hydrolysis
time
5hours

10 hours

15 hours

20 hours

25 hours

Enzim
Concentration
10%

Glucose (g)

20%

0.00672

30%

0.0096

40%

0.0144

10%

0.0192

20%

0.0298

30%

0.0388

40%

0.0468

10%

0.0468

20%

0.0488

30%

0.0518

40%

0.0588

In the study with the variation of the

weight of 50 grams with rice straw
hydrolysis time variations used were 5,
10, 15, 20 and 25 hours, and the
concentration of enzyme used 10%,
20%, 30% and 40% w / v, obtained
content glucose in each sample are
shown in Figure 1

0.00192

10%

0.0792

20%

0.08232

30%

0.0844

40%

0.08648

10%

0.08232

20%

0.0844

30%

0.0852

40%

0.08752

total enzyme fraction (5 ml enzyme per

50 grams of dry biomass) and close
tightly with a cork erlenmeyer. Then
placed on a rotary shaker at 160 rpm
for 25 hours. Glucose hydrolysis
results were analyzed by the method of
luff schoorl.

III. RESULTS AND DISCUSSION

3.1 Results
Glucose analysis results obtained in this
study of enzymatic hydrolysis process
results shown in Table 1, ie
Table 1. Results Analysis of Glucose
3.2 Discussion
a. Effect of Hydrolysis Time on Glucose

Figure 1. Effect of hydrolysis time on

glucose
Basically increasing hydrolysis time
can increase glucose levels generated
along with the many volumes of enzyme
were added. This is because the longer
the time given the more active side of
the enzyme cellulase work cut carbon
chains in the structure of cellulose into
simpler structures such as glucose or a
long time allowing the continuous
reaction of the enzyme to hydrolyze
cellulose to glucose. Vice versa, the less
time given to carry out the process of
hydrolysis eat the less cellulase enzyme
works to hydrolyze cellulose to glucose,
so glucose produced is also getting
smaller.
Seen from a glucose test results
obtained as in Figure 1 above. Glucose
test results did show a trend of increase
in glucose values with increasing
hydrolysis time. Of Figure 1 obtained the
highest glucose value of 0.08752 at 25
hours hydrolysis time.
b. Effect of Enzyme Concentration on
Glucose
Addition of enzyme concentration
on the amount of the fixed substrate,

generally increases the rate of glucose

production (Figure 2).

From the analysis of the Luff-Schoorl

method can obtain the highest glucose was
0.08752 gr at 40% addition of enzyme
concentration and hydrolysis time of 25
hours.

Acknowledgements
Authors would like to thank the
Higher Education research grant
through the National Strategic hisbah
2012.

Figure 2. Effect of enzyme concentration

on glucose
Can be seen in Figure 2, an increase in
glucose content of the addition of the
enzyme concentration of 10% to 40%
addition of enzyme concentration. For
example, the hydrolysis time 5 hours, an
increase in glucose were almost linearly
with increasing concentration of enzyme
added. The increase in glucose levels along
with the addition of large concentrations of
cellulase enzymes is because many levels of
cellulase enzymes resulted in the active site
of enzymes to break down cellulose into
glucose chains increase so that the enzyme
activity increases. The greater activity of
the enzyme, the more active side available
enzyme to break down cellulose into
glucose up.
The best concentration of enzyme to
hydrolyze rice straw is the addition of the
enzyme concentration of 40%. Therefore, it
can be concluded that the best 25-hour
hydrolysis time and enzyme concentrations
of 40% w / v glucose will be obtained at
0.08752 gr.
IV. CONCLUSION

REFERENCES
Binod, P., Sindhu.R, Singhania, R.R,
Vikram. S, Devi. L, Nagalakshmi. S.,
Kurien. N., Sukumaran, R K (2010),
Bioethanol Production from Rice
Straw, Bioresource Technology
101 : 4767- 4774
Cubero,
M.T.G.,
Benito,
G.,
Indacoechea, I.,Coca, M., Bolado, S
(2009), Effect of Ozonolysis
Pretreatment
on
Enzymatic
Digestibility of Wheat and Rye
Straw,
Bioresource Technology
100 :pp.1608 1613.
Fatma, H.A.E., dan Fadel, M (2010),
Production of Bioethanol Via
Enzymatic Saccharification of Rice
Straw by Cellulase Produced by
Tricodherma Reesei Under Solid
State Fermentation, New York
Science Journal : pp.72-78
Galleti, A.M R dan Antonetti, C. (2011),
Biomass Pre-treatment : Separation
of Cellulose, Hemicellulose and
Lignin. Existing Technologies and
Perspectives,
Department
of
Chemistry and Industrial Chemistry,
Universitas of Pisa, Italy.
Ong, L G.A., Chan, C. H dan Chew, A.L
(2012), Enzymatic Hydrolysis of

Rice Straw : Process Optimization,

Journal
of
Medical
and
Bioengineering (JOMB) Vol.1.pp.1416.

Taherzadeh, M.J dan Karimi, K.(2007),

Acid-Based Hydrolysis Processes for
Ethanol from Lignocellulosic Materials :
A Review, BioResources 2(3), pp : 472499

Roslam, A.M., Yee, P.L., Shah, U.K.M.,

Aziz, S.A dan Hassan, M.A (2011),
Production of Bioethanol from Rice
Straw Using Cellulase by Local
Aspergillus sp., International Journal
of Agricultural Research 6, pp :188193.

Yoswathana,
N.,
Phuriphipat,
P.,
Treyawutthiwat, P. and Naghi E. M.
(2010), Production Bioethanol from
Rice Straw, Energy Research Journal 1
(1) pp.26 31.