Enhancement of the enzymatic cellulose saccharification by Penicillium verruculosum multienzyme cocktails containing homologously overexpressed lytic polysaccharide monooxygenase

被引:0
作者
Margarita V. Semenova
Alexander V. Gusakov
Pavel V. Volkov
Veronika Yu. Matys
Vitaly A. Nemashkalov
Vadim D. Telitsin
Aleksandra M. Rozhkova
Arkady P. Sinitsyn
机构
[1] Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences,Department of Chemistry
[2] M. V. Lomonosov Moscow State University,G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms
[3] Russian Academy of Sciences,undefined
来源
Molecular Biology Reports | 2019年 / 46卷
关键词
Lytic polysaccharide monooxygenase; Homologous expression; Cellulase; Saccharification;
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学科分类号
摘要
The gene lpmo1 encoding Penicillium verruculosum lytic polysaccharide monooxygenase (PvLPMO9A) was sequenced and homologously overexpressed in P. verruculosum B1-537 (ΔniaD) auxotrophic strain under the control of the cbh1 gene promoter in combination with either the cbh1 signal sequence (sCBH1-X series of samples) or the native lpmo1 signal sequence (sLPMO1-X series). Three enzyme samples of the sCBH1-X series were characterized by a lower overall content of cellobiohydrolases (CBHs: 26–45%) but slightly higher content of endoglucanases (EGs: 17–23%) relative to the reference B1-537 preparation (60% of CBHs and 14% of EGs), while the PvLPMO9A content in them made up 9–21% of the total secreted protein. The PvLPMO9A content in four enzyme preparations of the sLPMO1-X series was much higher (30–57%), however the portion of CBHs in most of them (except for sLPMO1-8) decreased even to a greater extent (to 21–42%) than in the samples of the sCBH1-X series. Two enzyme preparations (sCBH1-8 and sLPMO1-8), in which the content of cellulases was substantially retained and the portion of PvLPMO9A was 9–30%, demonstrated the increased yields of reducing sugars in 48-h saccharification of Avicel and milled aspen wood: 19–31 and 11–26%, respectively, compared to the reference cellulase cocktail.
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页码:2363 / 2370
页数:7
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