期刊
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
卷 191, 期 2, 页码 463-481出版社
SPRINGER
DOI: 10.1007/s12010-019-03198-5
关键词
Thermophilic fungi; Heterologous expression; Lytic polysaccharide monooxygenases (LPMOs); H2O2; Hydrolysis
资金
- Department of Biotechnology, India [BT/PR15271/PBD/26/509/2015, BT/PR31115/PBD/26/766/2019]
- University of Houston
- State of Texas
In this study, two novel thermostable lytic polysaccharide monooxygenases (LPMOs) were cloned from thermophilic fungus Scytalidium thermophilum (PMO9D_SCYTH) and Malbranchea cinnamomea (PMO9D_MALCI) and expressed in the methylotrophic yeast Pichia pastoris X33. The purified PMO9D_SCYTH was active at 60 degrees C (t(1/2) = 60.58 h, pH 7.0), whereas, PMO9D_MALCI was optimally active at 50 degrees C (t(1/2) = 144 h, pH 7.0). The respective catalytic efficiency (k(cat)/K-m) of PMO9D_SCYTH and PMO9D_MALCI determined against avicel in presence of H2O2 was (6.58 x 10(-3) and 1.79 x 10(-3) mg(-1) ml min(-1)) and carboxy-methylcellulose (CMC) (1.52 x 10(-1) and 2.62 x 10(-2) mg(-1) ml min(-1)). The HRMS analysis of products obtained after hydrolysis of avicel and CMC showed the presence of both C-1 and C-4 oxidized oligosaccharides, in addition to phylogenetic tree constructed with other characterized type 1 and 3 LPMOs demonstrated that both LPMOs belongs to type-3 family of AA9s. The release of sugars during saccharification of acid/alkali pretreated sugarcane bagasse and rice straw was enhanced upon replacing one part of commercial enzyme Cellic CTec2 with these LPMOs.
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