Improvement of Saccharification and Delignification Efficiency of Trichoderma reesei Rut-C30 by Genetic Bioengineering

  • Gopalakrishnan, Raja Mohan; 
  • Manavalan, Tamilvendan; 
  • Ramesh, Janani; 
  • Thangavelu, Kalaichelvan Puthupalayam; 
  • Heese, Klaus
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초록

Trichoderma reesei produces various saccharification enzymes required for biomass degradation. However, the lack of an effective lignin-degrading enzyme system reduces the species' efficiency in producing fermentable sugars and increases the pre-treatment costs for biofuel production. In this study, we heterologously expressed the Ganoderma lucidum RMK1 versatile peroxidase gene (vp1) in the Rut-C30 strain of T. reesei. The expression of purified 6xHis-tag containing recombinant G. lucidum-derived protein (rVP1) was confirmed through western blot, which exhibited a single band with a relative molecular weight of 39 kDa. In saccharification and delignification studies using rice straw, the transformant (tVP7, T. reesei Rut-C30 expressing G. lucidum-derived rVP1) showed significant improvement in the yield of total reducing sugar and delignification, compared with that of the parent T. reesei Rut-C30 strain. Scanning electron microscopy (SEM) of tVP7-treated paddy straw showed extensive degradation of several layers of its surface compared with the parent strain due to the presence of G. lucidum-derived rVP1. Our results suggest that the expression of ligninolytic enzymes in cellulase hyperproducing systems helps to integrate the pre-treatment and saccharification steps that may ultimately reduce the costs of bioethanol production.

키워드

bioethanol; biomass degradation; delignification; Ganoderma lucidum; scanning electron microscope; Trichoderma reesei; versatile peroxidase; ERYNGII VERSATILE PEROXIDASE; HETEROLOGOUS PROTEIN EXPRESSION; WHITE-ROT FUNGUS; RICE STRAW; ENZYMATIC-HYDROLYSIS; TRANSFORMATION SYSTEM; PLEUROTUS-OSTREATUS; ETHANOL-PRODUCTION; FILAMENTOUS FUNGI; LACCASE GENE
제목
Improvement of Saccharification and Delignification Efficiency of Trichoderma reesei Rut-C30 by Genetic Bioengineering
저자
Gopalakrishnan, Raja Mohan; Manavalan, Tamilvendan; Ramesh, Janani; Thangavelu, Kalaichelvan Puthupalayam; Heese, Klaus
DOI
10.3390/microorganisms8020159
발행일
2020-02
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Article
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