International Journal of Modern Science and Technology
May 2019, Vol. 4, No. 5, pp 128-137.
A Novel Strain Developed through Protoplast Fusion for Consolidated Bioprocessing of Lignocellulosic Waste Mixture
R. Navnitkumar, T. R. Sambavi, S. Renganathan*
Biofuels Laboratory, Centre for Biotechnology, Anna University, Chennai – 600025. India.
*Corresponding author’s e-mail: email@example.com
A consolidated bioprocessing (CBP) strain is capable of producing cellulolytic enzymes to saccharify the lignocellulosic biomass and fermenting the produced monosaccharides to ethanol. In this work, protoplasts of Trichoderma harzanium ATCC® 20846™ and Saccharomyces cereviseae were obtained and they were fused using a PEG fusion method. The fused protoplasts were regenerated and screened for their cellulolytic activity and fermentative activity. An optimized Consolidated Bioprocessing was carried out for 160 hours using the fused protoplasts showing a regeneration frequency of 5 x 10-4. Around 0.04 g/L ethanol was produced by the CBP strain with a percentage cellulose conversion of 1.96%. SDS PAGE analysis was done to observe the proteins secreted by the CBP strain. The enzyme activities of the produced cellulase observed at the end of 160 hours were FPases: 0.9 IU/mL, CMCases: 6.30 IU/mL, Xylanases: 352.9 IU/mL and Beta-glucosidases 1491.2 IU/mL.
Keywords: Protoplast fusion; Consolidated bioprocessing; Lignocellulosic waste; Regeneration frequency; Cellulase.
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