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8/3/2019 Utilization of Cellulosic Bio Resource
http://slidepdf.com/reader/full/utilization-of-cellulosic-bio-resource 1/23
Utilization of cellulosicUtilization of cellulosic
bioresourcebioresource CelluloseCellulose -- a major source renewablea major source renewable
of carbon sourceof carbon source
Difficult to utilizeDifficult to utilize
Technology availableTechnology available economical?economical?
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Major sources ofMajor sources of
cellulosicscellulosics Palm oil processingPalm oil processing
Rice productionRice production Timber processingTimber processing
Manucipal solid wastesManucipal solid wastes
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Cellulosics from palm oilCellulosics from palm oil
processingprocessing
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Cellulosics from palm oilCellulosics from palm oil
processingprocessing
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Palm biomassPalm biomass carboncarbon
exploitationexploitation
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Palm biomassPalm biomass aa
renewable resourcerenewable resource
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Production of PHB fromProduction of PHB from
POMEPOME
Use lipolytic organismsUse lipolytic organisms produceproducelipases to breakdown lipids to fattylipases to breakdown lipids to fattyacids and glycerolacids and glycerol
Fatty acids then broken down throughFatty acids then broken down through F F--oxidation to produce acetyl CoAoxidation to produce acetyl CoA
Acetyl CoA produced incorporated into Acetyl CoA produced incorporated intoPHBPHB
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Pathway for production of PHB Pathway for production of PHB using palm oil (Toh Mong Sahusing palm oil (Toh Mong Sah et alet al..
1995)1995)
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Screening and isolation of PHA Screening and isolation of PHA
producers ( Amirulproducers ( Amirul et alet al. 2007). 2007)Soil samplesSoil samples
663 isolates663 isolates
Nile red stainingNile red staining
119 isolates were identified as possible PHA producers119 isolates were identified as possible PHA producers
cultivated in MSM containcultivated in MSM contain --butyrolactonebutyrolactone
95 isolates confirmed by GC analysis95 isolates confirmed by GC analysis
77 isolates produced homopolymer77 isolates produced homopolymer 18 isolates produced copolymer18 isolates produced copolymer
P(3HB)P(3HB) P(3HBP(3HB--coco--4HB)4HB)
USMAA1020USMAA1020
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Observation PHA producersObservation PHA producers
under UV light microscopeunder 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.
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Characterization of isolate USMAA1020Characterization of isolate USMAA1020
Morphological and biochemical
test resulted ;
Gram-negative
Aerobic
Non-spore forming
Motile
Rod-shaped
(0.7 ± 0.9 µm x 1.53 ± 3.0 µm)
Assimilates p-hydroxy-benzoate,
mesaconate, trans-aconitate,D/L-tartrate, citrate, malate,
adipate, caprate and gluconate
Growth at 30ºC and
37ºC but not at 42ºC
DNA G + C content is 62.3%
These results suggested that USM AA1020 belongs to the genus
Cupriavidus (Vandamme and Coenye, 2004).
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Morphology of the isolateMorphology of the isolate
USMAA1020USMAA1020
Transmission electron micrograph of USM AA1020.
Morphology of cell with no PHA accumulation
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TEM of USM AA 1020 showing morphology of cell with PHA granules in
the cell cytoplasm. PHA granules appear as electron-dense inclusions.
Morphology of the isolate USMAA1020Morphology of the isolate USMAA1020
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Strategy for utilization ofStrategy for utilization of
cellulosicscellulosics Use enzyme cellulase for breakdownUse enzyme cellulase for breakdown
to produce glucoseto produce glucose
CellulaseCellulase a complex of 3 enzymesa complex of 3 enzymes
cellobiohydrolasecellobiohydrolase
endoglucanaseendoglucanase
F F--glucosidaseglucosidase
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Sugar production fromSugar production from
cellulosicscellulosics Enzymes very expensiveEnzymes very expensive
PrePre--treatment of cellulosics required totreatment of cellulosics required tofacilitate hydrolysisfacilitate hydrolysis
Sugars produced can be utilized forSugars produced can be utilized forfermentation of various productsfermentation of various products
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Mushroom productionMushroom production
A viable way to utilize cellulosics A viable way to utilize cellulosics
Has been shown to be commerciallyHas been shown to be commerciallyviableviable
Spent substrate can be used asSpent substrate can be used asfertilizerfertilizer
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What are mushrooms?What are mushrooms?
Mushroom is a simple form of lifeMushroom is a simple form of lifeknown as fungusknown as fungus
Mushrooms are the edible fleshyMushrooms are the edible fleshyfruiting bodies of certain fungifruiting bodies of certain fungi
Mushrooms are popular for theirMushrooms are popular for their
delicacy and flavoured food valuedelicacy and flavoured food value Gathered wild or grown underGathered wild or grown under
cultivationcultivation
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Varieties of mushrooms Varieties of mushrooms
commercially cultivatedcommercially cultivated Oyster mushroom (Oyster mushroom (PleurotusPleurotus sp.)sp.)
Paddy straw mushroomPaddy straw mushroom (Volvariella(Volvariellaspp.)spp.)
Shiitake (Shiitake (Lentinus edodesLentinus edodes))
Button mushroom (Button mushroom (Agaricus bisporus Agaricus bisporus))
Maitake (Maitake (Gifola frondosaGifola frondosa))
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Steps in mushroomSteps in mushroom
cultivationcultivation 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
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Oyster mushroomOyster mushroom
cultivation flowcultivation flow--chartchartRaw material Chopping (3-5 cm) Soak straw (6-18 h)
Drain off excesswater
Fill-up gunnybags
Dip in hot water80-85oC x 15 min
Drain off excess
water
When cool to roomtemperature, fill bags in 3
layers of 10 cm each andsimultaneously spawn @2% inside sterile chamber
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Oyster mushroomOyster mushroom
cultivation flowcultivation flow--chartchart Arrange bags in shelves(Spawn running requires20-25 days)
Pin head stage (takes 3-5days)
Cropping, 1st flush (3-4 daysduration after pin head ) andsubsequent flushes about 10days apart
Plucking on maturity(before spore shed starts)
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What are potentialWhat are potential
substrates?substrates? Rice strawRice straw
Palm fibres?Palm fibres?
Saw dust?Saw dust?
Wood chips?Wood chips?
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ReferencesReferences
MIAMIA Majid,Majid, DHDH Akmal, Akmal, LLLL FewFew.. A A Agustien, Agustien, MSMS Toh,Toh, MRMRSamian,Samian, NN NajimudinNajimudin andand MNMN Azizan Azizan ((19991999)).. ProductionProduction of of poly(poly(--33--hydroxybutyrate)hydroxybutyrate) byby ErwiniaErwinia spsp.. USMIUSMI--2020.. Int Int..
JournalJournal of of BiologicalBiological MacromoleculesMacromolecules 2525:: 9595--104104.. TohToh MongMong Sah,Sah, IsaIsa Abd Abd.. MajidMajid andand Azizan Azizan MohdMohd.. NoorNoor
((19951995)).. Aspects Aspects of of poly(poly(33--hydroxybutyrate)hydroxybutyrate) productionproduction bybyErwiniaErwinia spsp.. usingusing palmpalm oiloil asas thethe substratesubstrate.. 1818thth MalaysianMalaysianMicrobiologyMicrobiology SymposiumSymposium.. 2323--2424 August, August, Kuching,Kuching, SarawakSarawak..