›› 2009, Vol. 11 ›› Issue (4): 64-70.

• 863课题进展 • 上一篇    下一篇

链霉菌Streptomyces sp. S9木聚糖酶基因xynBS9的克隆表达及性质分析

姚国玉1,2,李宁2,石鹏君2,陈强1,姚斌2   

  1. 1.兰州大学生命科学学院生物化学与分子生物学研究所,兰州 730000;2.中国农业科学院饲料研究所农业部饲料生物技术重点实验室,北京 100081
  • 收稿日期:2009-04-14 修回日期:2009-06-15 出版日期:2009-08-15 发布日期:2009-07-24
  • 通讯作者: 姚斌,研究员,博士,博士生导师,从事微生物工程研究。Tel:010-82106053; E-mail:yaobin@mail.caas.net.cn。陈强,教授,博士,博士生导师,从事生物化学与分子生物学研究。E-mail:chenq@lzu.edu.cn
  • 作者简介:姚国玉,硕士研究生,从事微生物工程研究。E-mail:gyyao666@163.com
  • 基金资助:

    国家863计划项目(2007AA100601);国家科技攻关项目(2006BAD12B05-03)资助。

Cloning, Expression and Characterization of a Xylanase Gene, xynBS9, from Streptomyces sp. S9

YAO Guo-yu1, LI Ning2, SHI Peng-jun2, CHEN Qiang1,YAO Bin2   

  1. 1.Institute of Biochemistry and Molecule Biology, School of Life Science, Lanzhou University, Lanzhou 730000|2.Key Laboratory of Feed Biotechnology, Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2009-04-14 Revised:2009-06-15 Online:2009-08-15 Published:2009-07-24

摘要:

通过设计简并引物和构建基因组文库的方法从链霉菌Streptomyces sp.S9中克隆得到β-l,4-木聚糖酶基因xynBS9。该基因全长1 023 bp,编码340个氨基酸。将不带原基因信号肽编码序列的xynBS9以正确阅读框架克隆到表达载体pET-22b(+)上,并在大肠杆菌BL2l(DE3)中诱导表达。重组蛋白经硫酸铵分级沉淀和疏水柱纯化后达到电泳纯。酶学性质分析表明,重组木聚糖酶最适温度为60℃,最适pH为6.5,在碱性条件下具有良好的稳定性。

关键词: 链霉菌;木聚糖酶;xynBS9基因;原核表达

Abstract:

The gene xynBS9 encoding β-l, 4-xylanase was cloned from Streptomyces sp.S9 by designing degenerate primers and screening from a genomic library of Streptomyces sp. S9. The xynBS9 gene was 1 023 bp in length and encoded by a protein with 340 amino acids. The xynBS9 gene without signal peptide was inserted into the expression vector of pET-22b (+) and transformed into Escherichia coli BL21 (DE3) to express. The recombinant protein was purified by ammonium sulfate precipitant and hydrophobic interaction chromatography.Characteristic analysis indicated that the optimum temperature and pH value for the recombined xylanase were 60℃ and 6.5, respectively, and XynBS9 showed an extreme stability under alkaline condition.

Key words: Streptomyces, xylanase, xynBS9 gene, prokaryotic expression

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