EVALUATION OF DIFFERENT PHYSICO-CHEMICAL AND BIOLOGICAL PRETREATMENTS FOR ENZYMATIC HYDROLYSIS OF HARDWOOD FOR ETHANOL PRODUCTION
KrishiKosh
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Title |
EVALUATION OF DIFFERENT PHYSICO-CHEMICAL AND BIOLOGICAL PRETREATMENTS FOR ENZYMATIC HYDROLYSIS OF HARDWOOD FOR ETHANOL PRODUCTION
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Creator |
KAUSHAL, RICHA
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Contributor |
SHARMA, NIVEDITA
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Subject |
enzymes, productivity, fungi, fermentation, cellulose, bacteria, wood, polysaccharides, alcohols, poultry equipment
Hardwood , Potential Microorganisms |
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Description |
ABSTRACT In the present investigation, an attempt was made to utilize hardwood as substrate for its degradation by potential microorganisms and evaluated different pretreatments to enhance their rate of hydrolysis - a key step for its bioconversion to ethanol. In total 20 microorganisms including 17 bacteria and 3 fungi were isolated. Among them, SD5 and RS2 were screened for cellulase and SD8 for xylanase production and were identified as B. simplex SD5, B. subtilis RS2, B. subtilis SD8 by 16S rRNA PCR technique and registered with NCBI under accession no KF844070, KF844069 and KF844068, respectively. Among fungi, WF5 and RF1 were selected for enzyme production and were identified using ITS 5.8S rRNA technique as T. harzianum WF5 and R. oryzae RF1 and registered under accession no. KF844067 and KJ1921199, respectively. Cellulase and xylanase enzymes were optimized through classical approach one factor at a time (OFAT) and Response surface methodology (RSM) varying medium, pH, temperature, inoculum size, incubation time, and substrate concentration. The partial purification of hydrolytic enzymes was done by ammonium sulphate precipitation. Full length gene sequences of BsSD8-xylanase of B. subtilis SD8 and four GHs namely three subunits of cellulase, Endoglucanase (ThWF5-Endo-glucanase), Exo-glucanase (ThWF5-Exo-glucanase) and -glucosidase (ThWF5- Glucosidase), and xylanase (ThWF5-Xylanase) of T. harzianum WF5 were pulled out and characterized. To reduce the production cost of ethanol, cheap untreated and pretreated lignocellulosic forest biomass i.e. hardwood were used as a substrate for sugar production. Among different hardwood species used, Eucalyptus and P. deltoides wood were selected for saccharification by bacterial and fungal hydrolytic enzymes, respectively. Among different physical, chemical and biological pretreatments, H2SO4 + H2O2 + steam pretreatment was found best for sugar production. Bioconversion of H2SO4 + H2O2 + steam pretreated E. teretecornis and P. deltoides wood to ethanol was studied under two different fermentation processes i.e separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). Different protocols had been designed to delimit the constraints of fermentation process. SHF was evaluated by modifying four different sub-processes of non-detoxification and detoxification as well as nonpooling and pooling of pretreated liquor. Maximum ethanol was achieved in protocol IV i.e. 7.02 g/l by co-culture of S. cerevisiae I + P. stipitis in E. teretecornis wood and 15.62 g/l in P. deltoides wood with fermentation efficiency of 61.05%. Scale up of SHF with P. deltoides wood using fungal enzymes and co-culture of S. cerevisiae I + P. stipitis was performed in 7.5 L bioreactor, achieving highest ethanol production after 52 h of fermentation. Among SHF and SSF, SHF in protocol IV i.e. pooled sample followed by detoxification was found to be the best and in case of strains used for fermentation, co-culture of S. cerevisiae I + P. stipitis was observed the best combination for highest bioethanol production |
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Date |
2016-06-16T10:57:43Z
2016-06-16T10:57:43Z 2015 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/67443
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Language |
en
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Format |
application/pdf
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