A Novel Strain Developed through Protoplast Fusion for Consolidated Bioprocessing of Lignocellulosic Waste Mixture

Abstract

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 h 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 h 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

References

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