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假定甲基转移酶过表达对用于生物质转化的哈茨木霉纤维素分解系统的影响。

The impact of putative methyltransferase overexpression on the Trichoderma harzianum cellulolytic system for biomass conversion.

机构信息

National Centre of Research in Energy and Materials, High-tech Pole II, 10000 Giuseppe Maximo Scolfaro St, P.O Box 6192, Campinas, SP, Brazil; Synthetic Biology and Molecular Biotechnology, Research Division Biochemical Technology, Institute of Chemical Engineering, TU Wien, Gumpendorferstrasse 1a, A-1060 Vienna, Austria; Federal University of Goiás (UFG), Samambaia Campus, Goiânia, GO, Brazil.

National Centre of Research in Energy and Materials, High-tech Pole II, 10000 Giuseppe Maximo Scolfaro St, P.O Box 6192, Campinas, SP, Brazil.

出版信息

Bioresour Technol. 2020 Oct;313:123616. doi: 10.1016/j.biortech.2020.123616. Epub 2020 Jun 14.

Abstract

Trichoderma harzianum has attracting attention for its potential alternative use in biofuel production, due to a recognized competence for high diversity glycoside hydrolases (GH) enzyme complex, including higher β-glucosidases and auxiliary proteins, using low-cost carbon sources. This strain constitutively overexpressed the global regulator putative methyltransferase - LAE1, in order to improve the GHs production. The recombinant strain achieved 79-fold increase in lae1 expression and high GHs productivity. The evaluation of the LAE1 impact to induce the GHs used soluble and lignocellulose inexpensive carbon sources in a stirred-tank bioreactor. Using sugarcane bagasse with sucrose, the overexpression of lae1 resulted in significantly increment of gh61b (31x), cel7a (25x), bgl1(20x) and xyn3 (20x) genes expression. Reducing sugar released from pretreated sugarcane bagasse, which hydrolyzed by recombinant crude enzyme cocktail, achieved 41% improvement. Therefore, lae1 overexpression effectively is a promising improving GHs target for biomass degradation by T. harzianum.

摘要

哈茨木霉因其在生物燃料生产中的潜在替代用途而受到关注,因为它具有公认的使用低成本碳源生产高多样性糖苷水解酶(GH)酶复合物的能力,包括更高的β-葡萄糖苷酶和辅助蛋白。该菌株组成型过表达全局调节剂假定甲基转移酶-LAE1,以提高 GHs 的产量。重组菌株实现了 lae1 表达的 79 倍增加和高 GHs 生产力。在搅拌罐生物反应器中使用可溶性和木质纤维素廉价碳源评估 LAE1 对诱导 GHs 的影响。使用甘蔗渣和蔗糖,lae1 的过表达导致 gh61b(31x)、cel7a(25x)、bgl1(20x)和 xyn3(20x)基因表达显著增加。用重组粗酶混合物水解预处理过的甘蔗渣释放的还原糖,提高了 41%。因此,lae1 的过表达是一种很有前途的方法,可以有效地提高哈茨木霉对生物质降解的 GHs 目标。

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