疏绵状嗜热丝孢菌外切β-葡聚糖酶的基因克隆与酶学特征

发布时间：2018-01-08 14:25

本文关键词：疏绵状嗜热丝孢菌外切β-葡聚糖酶的基因克隆与酶学特征　出处：《微生物学通报》2016年02期 　论文类型：期刊论文

【摘要】：【目的】疏绵状嗜热丝孢菌是一种嗜热丝状真菌,具有合成与分泌多种耐热酶的能力,从中找寻酶活高、耐热性能优良的β-葡聚糖酶。【方法】对疏绵状嗜热丝孢菌其中一个外切β-葡聚糖酶的编码基因gln B进行克隆,并在毕赤酵母GS115中表达。【结果】在摇瓶水平上重组菌的产酶活为11.5 U/m L,重组酶在SDS-PAGE中的大小约为48 k D。重组Gln B最适作用温度为65°C,最适作用p H为5.0;在低于50°C或p H 3.0-10.0之间具有良好稳定性。重组酶水解海带三糖,首先生成单糖和二糖,延长酶作用时间,可以进一步将其中的二糖部分水解为单糖。【结论】疏绵状嗜热丝孢菌来源的重组酶Gln B为外切β-葡聚糖酶,具有较好的耐热性能和p H稳定性。
[Abstract]:[objective] Trichoderma thermophilus is a thermophilic filamentous fungus, which has the ability to synthesize and secrete a variety of heat-resistant enzymes. [methods] one of the encoding genes of 尾 -glucanase gln B was cloned. And expressed in Pichia pastoris GS115. [results] the enzyme production activity of the recombinant strain was 11.5 UmL at shaking flask level. The size of recombinant enzyme in SDS-PAGE was about 48 KD, the optimum temperature and pH of recombinant Gln B were 65 掳C and 5.0, respectively. It has good stability between 50 掳C or pH 3.0-10.0.Recombinant enzyme hydrolyzed kelp trisaccharide, first of all, monosaccharide and disaccharide were formed, and the reaction time of enzyme was prolonged. [conclusion] the recombinant enzyme Gln B, from which the disaccharide was partially hydrolyzed to monosaccharide, was a 尾 -glucanase with good heat resistance and pH stability.
【作者单位】：天津科技大学生物工程学院;Department
【基金】：国家自然基金国际(地区)合作与交流项目(No.31461143026) 天津市应用基础与前沿技术研究计划(青年)项目(No.14JCQNJC09200)~~
【分类号】：Q936;Q78
【正文快照】： (2.Department of BiotechnologyFood Technology,Faculty of Applied Sciences,Durban University of Technology,P.O.Box 1334,Durban,4001,South Africa)(3.福州大学生物科学与工程学院福建福州350116)β-葡聚糖以右旋葡萄糖为基本单位,通过β-1,3-(1,4)-糖苷键聚合而成,

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