[1]李 飞,解静聪,李 琦,等.木聚糖酶XynB分子Ser/Thr 平面导入精氨酸对酶热稳定性的影响[J].南京林业大学学报(自然科学版),2014,38(04):107-112.[doi:10.3969/j.issn.1000-2006.2014.04.020]
　LI Fei,XIE Jingcong,LI Qi,et al.Thermostability of xylanase(XynB)by introducing Arg into its Ser/Thr surface[J].Journal of Nanjing Forestry University(Natural Science Edition),2014,38(04):107-112.[doi:10.3969/j.issn.1000-2006.2014.04.020]
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木聚糖酶XynB分子Ser/Thr 平面导入精氨酸对酶热稳定性的影响
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《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

卷:
38
期数:
2014年04期
页码:
107-112
栏目:
研究论文
出版日期:
2014-07-01

文章信息/Info

Title:
Thermostability of xylanase(XynB)by introducing Arg into its Ser/Thr surface
文章编号:
1000-2006(2014)04-0107-06
作者:
李 飞1; 2; 解静聪1; 李 琦1; 张雪松4; 赵林果1;  3*
1.南京林业大学化学工程学院, 江苏 南京 210037;
2.南京师范大学泰州学院生物技术与化学工程学院, 江苏 泰州 225300;
3.江苏省生物质绿色燃料与化学品重点实验室,江苏 南京 210037;
4.江苏农林职业技术学院生物工程系,江苏 镇江 212400
Author(s):
LI Fei1; 2;  XIE Jingcong1; LI Qi1; ZHANG Xuesong4; ZHAO Linguo1; 3*
1.College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China;
2.College of Biochemical Engineering, Taizhou College, Nanjing Normal University, Taizhou 225300, China;
3.Jiangsu Key Lab of Biomass Based Green Fuels and Chemicals, Nanjing 210037, China;
4.Department of Biological Engineering, Jiangsu Polytechnic College of Agriculture and Forestry, Zhenjiang 212400, China
关键词:
定点突变;  木聚糖酶;  精氨酸;  热稳定性
Keywords:
site-directed mutagenesis;  xylanase;  arginine;  thermostability
分类号:
TQ936
DOI:
10.3969/j.issn.1000-2006.2014.04.020
文献标志码:
A
摘要:
为提高木聚糖酶的热稳定性,通过定点突变的方法,在来源于黑曲霉Aspergillus niger nl-1的木聚糖酶XynB分子Ser/Thr 平面上引入5个精氨酸,实现了突变酶在毕赤酵母中的表达。通过酶学性质的研究表明,突变酶XynB-104和XynB-77的最适反应温度为50 ℃,且与原酶相比相对酶活力得到显著提高。在存在底物的情况下,50 ℃热处理2 h,突变酶XynB-104和XynB-77残余酶活力较原酶的相对转化效率由26%提高到80%左右。突变酶XynB-104的最适反应pH与原酶一致,而突变酶XynB-77最适pH由5.0提高到5.5。结果表明,在木聚糖酶分子Ser/Thr 平面不同折叠片引入精氨酸可以增强酶结构的稳定性,特别是能显著提高热稳定性。
Abstract:
Replacing several serine and threonine residues in the Ser/Thr surface of xylanase(XynB)with arginines effectively increased the thermostability of the enzyme by site-directed mutagenesis. The mutated enzymes XynB-104 and XynB-77 were expressed in Pichia pastoris and their enzymatic properties were determined. The optimal temperature of the mutations XynB-104 and XynB-77 were 50 ℃, and the relative activity of mutations were dramatically increased. Both mutated xylanases showed about 80% of maximal activity after incubating in xylan substrate for 2 h at 50 ℃ compared to only 26% activity of the native enzyme. The optimal pH of XynB-104 enzyme had no change, but the pH optimum of the XynB-77 enzyme was increased from 5.0 to 5.5. The results of the mutated enzymes indicated that properly arginines residues introduced into Ser/Thr surface of xylanase XynB might be effective to improve the enzyme thermostability and hydrolysis capacity.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2013-01-10 修回日期:2013-10-21
基金项目:江苏省高校自然科学研究重大项目(13KJA220004); 江苏省普通高校研究生科研创新计划资助项目(CXLX12_0529); 江苏高校优势学科建设工程资助项目(PAPD)
第一作者:李飞,讲师。*通信作者:赵林果,教授。E-mail: lg.zhao@163.com。
引文格式:李飞,解静聪,李琦,等. 木聚糖酶XynB分子Ser/Thr 平面导入精氨酸对酶热稳定性的影响[J]. 南京林业大学学报:自然科学版,2014,38(4):107-112.
更新日期/Last Update: 2014-07-31