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Utilization of cellulosic bioresource


Cellulose - a major source renewable of carbon source Difficult to utilize Technology available economical?

Major sources of cellulosics


Palm oil processing Rice production Timber processing Manucipal solid wastes

Cellulosics from palm oil processing

Cellulosics from palm oil processing

Palm biomass exploitation


Palm biomass a renewable resource

Production of PHB from POME

Use lipolytic organisms produce lipases to breakdown lipids to fatty acids and glycerol Fatty acids then broken down through F-oxidation to produce acetyl CoA Acetyl CoA produced incorporated into PHB

Pathway for production of PHB using palm oil (Toh Mong Sah et al. al. 1995)

Screening and isolation of PHA producers (Amirul et al. 2007) al.

Soil samples 663 isolates
Nile red staining

119 isolates were identified as possible PHA producers

cultivated in MSM contain -butyrolactone

95 isolates confirmed by GC analysis 77 isolates produced homopolymer P(3HB) 18 isolates produced copolymer P(3HB-coP(3HB-co-4HB)


Observation PHA producers under UV light microscope

Individual cells when observed under UV light microscope. PHA granules in the cell cytoplasm emitted orange fluorescence when exposed to UV light.

Characterization of isolate USMAA1020

Aerobic Gram-negative Rod-shaped (0.7 0.9 m x 1.53 3.0 m) Motile Morphological and biochemical test resulted ; Non-spore forming

Growth at 30C and 37C but not at 42C

DNA G + C content is 62.3%

Assimilates p-hydroxy-benzoate, mesaconate, trans-aconitate, D/L-tartrate, citrate, malate, adipate, caprate and gluconate

These results suggested that USMAA1020 belongs to the genus Cupriavidus (Vandamme and Coenye, 2004).

Morphology of the isolate USMAA1020

Transmission electron micrograph of USMAA1020. Morphology of cell with no PHA accumulation

Morphology of the isolate USMAA1020

TEM of USMAA 1020 showing morphology of cell with PHA granules in the cell cytoplasm. PHA granules appear as electron-dense inclusions.

Strategy for utilization of cellulosics

Use enzyme cellulase for breakdown to produce glucose Cellulase a complex of 3 enzymes
cellobiohydrolase endoglucanase F-glucosidase

Sugar production from cellulosics


Enzymes very expensive PrePre-treatment of cellulosics required to facilitate hydrolysis Sugars produced can be utilized for fermentation of various products

Mushroom production

A viable way to utilize cellulosics Has been shown to be commercially viable Spent substrate can be used as fertilizer

What are mushrooms?


Mushroom is a simple form of life known as fungus Mushrooms are the edible fleshy fruiting bodies of certain fungi Mushrooms are popular for their delicacy and flavoured food value Gathered wild or grown under cultivation

Varieties of mushrooms commercially cultivated



Oyster mushroom (Pleurotus sp.) (Pleurotus Paddy straw mushroom (Volvariella spp.) Shiitake (Lentinus edodes) (Lentinus edodes) Button mushroom (Agaricus bisporus) (Agaricus bisporus) Maitake (Gifola frondosa) (Gifola frondosa)

Steps in mushroom cultivation


Choosing a growing medium ie. substrate Pasteurizing or sterilizing the medium Seeding the beds with spawn Maintaining optimal temperature, moisture, and other conditions for mycelium growth and the conditions that favor fruiting - most challenging step Harvesting, packaging, and marketing

Oyster mushroom cultivation flow-chart flowRaw material Chopping (3-5 cm) Soak straw (6-18 h) Dip in hot water 80-85oC x 15 min Fill-up gunny bags Drain off excess water

Drain off excess water

When cool to room temperature, fill bags in 3 layers of 10 cm each and simultaneously spawn @ 2% inside sterile chamber

Oyster mushroom cultivation flow-chart flowArrange bags in shelves (Spawn running requires 20-25 days) Pin head stage (takes 3-5 days)

Plucking on maturity (before spore shed starts)

Cropping, 1st flush (3-4 days duration after pin head ) and subsequent flushes about 10 days apart

What are potential substrates?


Rice straw Palm fibres? Saw dust? Wood chips?


MIA Majid, DH Akmal, LL Few. A Agustien, MS Toh, MR Few. Samian, N Najimudin and MN Azizan (1999). Production of 1999) poly(poly(-3-hydroxybutyrate) by Erwinia sp. USMI-20. Int. sp. USMI-20. Int. Journal of Biological Macromolecules 25: 95-104. 25: 95-104. Toh Mong Sah, Isa Abd. Majid and Azizan Mohd. Noor Abd. Mohd. (1995). Aspects of poly(3-hydroxybutyrate) production by 1995) poly(3 Erwinia sp. using palm oil as the substrate. 18th Malaysian sp. substrate. 18th Microbiology Symposium. 23-24 August, Kuching, Sarawak. Symposium. 23Sarawak.