首页> 外文期刊>Biotechnology Letters >Identification of two novel xylanase-encoding genes (xyn5 and xyn6) from Acrophialophora nainiana and heterologous expression of xyn6 in Trichoderma reesei
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Identification of two novel xylanase-encoding genes (xyn5 and xyn6) from Acrophialophora nainiana and heterologous expression of xyn6 in Trichoderma reesei

机译:鉴定两个新的木霉编码木聚糖酶编码基因(xyn5和xyn6)和里氏木霉中xyn6的异源表达

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摘要

Two novel genes,xyn5 and xyn6,coding for family 11 xylanases,were isolated from the thermotolerant filamentous fungus,Acrophialophora nainiana,by PCR using degenerate primers.The xyn6 gene was further expressed in Trichoderma reesei.DNA sequence analysis of xyn6 revealed an open reading frame (ORF) of 708 bp,interrupted by an intron of 58 bp.The xyn6 ORF encodes a predicted protein of 236 amino acid residues.The mature recombinant XynVI protein had a molecular mass of about 19 kDa,as estimated by gel electrophoresis.Analysis of the predicted amino acid sequence of XynVI paves the way for rational protein engineering by site-directed mutagenesis aiming to increase the thermostability of the heterologously-expressed xylanase.
机译:利用简并引物通过PCR从耐热丝状真菌Acrophialophora nainiana中分离出两个编码11家族木聚糖酶的新基因xyn5和xyn6。xyn6基因在里氏木霉中进一步表达。xyn6的DNA序列分析揭示了开放阅读框架(ORF)为708 bp,被58 bp的内含子打断.xyn6 ORF编码一个236个氨基酸残基的预测蛋白。通过凝胶电泳估计,成熟的重组XynVI蛋白的分子质量约为19 kDa。 XynVI的预测氨基酸序列的确定为通过旨在增加异源表达的木聚糖酶的热稳定性的定点诱变铺平了合理的蛋白质工程道路。

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