米曲霉木聚糖酶N端引入二硫键对其热稳定性的影响

doi:10.3969/j.issn.1673-1689.2013.07.003

首页 > 过刊浏览>2013年第32卷第7期 >686-691. DOI:10.3969/j.issn.1673-1689.2013.07.003

米曲霉木聚糖酶N端引入二硫键对其热稳定性的影响
DOI:
                        10.3969/j.issn.1673-1689.2013.07.003
                    
作者:
                        
                        
                    
作者单位:
作者简介:
通讯作者:
中图分类号:
基金项目:国家自然科学基金项目(31101229)

Effect of Engineering a Disulfide Bridge into the N-terminal Region of Aspergillus oryzae Xylanase on its Thermostability

Author:

Affiliation:

Fund Project:

摘要

图/表

访问统计

参考文献

相似文献

引证文献

资源附件

文章评论

摘要:

为提高源自米曲霉Aspergillus oryzae 11家族木聚糖酶(AoXyn11A)的热稳定性,对其相关的氨基酸残基进行定点突变改造。通过对野生型酶AoXyn11A和一种超耐热酶EvXyn11TS的N端氨基酸残基序列进行同源比较,在野生型酶N端引入一个二硫键,获得突变酶AoXyn11AM。野生型酶和突变酶的热稳定性通过同源建模和分子动力学模拟分析评估后,其基因分别在毕赤酵母GS115中进行表达并分析温度对表达产物酶活性的影响。结果表明:突变酶的最适温度由野生型酶的55℃提高至60℃;在50℃和55℃保温30 min,突变酶保留94%和45%保温处理前的酶活性,分别较野生型酶(62.5%和1.4%)有大幅度的提高。突变酶在保留了野生酶其它优良性质的基础上,提高了热稳定性,具有潜在的工业应用价值。

Abstract:

Site-directed mutagenesises had been done in this work with the consideration of increasing the thermostability of a family 11 xylanase(AoXyn11A) from Aspergillus oryzae.The mutant,AoXyn11AM,was obtained by introducing a disulfide bridge into the N-terminal region of AoXyn11A after N-terminal amino acid homology alignment of AoXyn11A with a thermostable family 11 xylanase,EvXyn11TS.The thermostability of the wild-type enzyme and mutant were analyzed by homology modeling,and assessed by molecular dynamics(MD) simulations.The genes,AoXyn11A and AoXyn11AM,were expressed in Pichia pastoris GS115,respectively.And the effects of temperatures on the thermostability of the expressed AoXyn11A and AoXyn11AMwere analyzed.The results showed the optimal temperature of AoXyn11AMwas raised to 60 ℃ from 55 ℃ of AoXyn11A;after incubated at 50 ℃ and 55 ℃ for 30 min,respectively,the mutant retained 94% and 45% of its original activity while the wild-type enzyme retained 62.5% and 1.4% of its original activity.The mutant not only preserved the major excellent properties of the wild-type enzyme,but also enhanced the thermostability which is potentially useful in industrial applications.

参考文献

相似文献

引证文献

引用本文

陈忠法,唐存多,汪俊卿,吴静,邬敏辰.米曲霉木聚糖酶N端引入二硫键对其热稳定性的影响[J].食品与生物技术学报,2013,32(7):686-691.

CHEN Zhong-fa, TANG Cun-duo, WANG Jun-qing, WU Jing, WU Min-chen. Effect of Engineering a Disulfide Bridge into the N-terminal Region of Aspergillus oryzae Xylanase on its Thermostability[J]. Journal of Food Science and Biotechnology,2013,32(7):686-691.

复制

文章指标

点击次数:
下载次数:
HTML阅读次数:
引用次数:

历史

收稿日期:
最后修改日期:
录用日期:
在线发布日期: 2014-06-17
出版日期:

地址：江苏省无锡市蠡湖大道1800号邮政编码：214122

电话：0510-85913526 电子邮件：xbbjb@jiangnan.edu.cn

技术支持：北京勤云科技发展有限公司

引用本文

分享

文章指标

历史

微信公众号二维码

手机版网站二维码