Studies on Cloning of Acid Xylanase from Phialophora sp. G5 and its Characteristics

doi:10.3969/j.issn.1008-0864.2011.03.10

›› 2011, Vol. 13 ›› Issue (3): 60-66.DOI: 10.3969/j.issn.1008-0864.2011.03.10

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Studies on Cloning of Acid Xylanase from Phialophora sp. G5 and its Characteristics

ZHANG Fan1,2, SHI Peng-jun2, MIAO Li-hong1, YAO Bin2

(1.School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023|2.Key Laboratory for Feed Biotechnology, Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China )

Received:2011-02-22 Revised:2011-04-09 Online:2011-06-15 Published:2011-06-16

来源于瓶霉Phialophora sp. G5的酸性木聚糖酶基因的克隆及性质的研究

张帆1,2,石鹏君2,缪礼鸿1,姚斌2

(1.武汉工业学院生物与制药工程学院, 武汉 430023|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)

通讯作者: 缪礼鸿,教授,博士,硕士生导师,研究方向为资源与环境微生物学。E-mail:lhmiao2002@yahoo.com.cn；姚斌,研究员,博士,博士生导师,研究方向为微生物基因工程。E-mail: yaobin@caas-bio.net.cn
作者简介:张帆,硕士研究生,从事微生物基因工程研究。E-mail:zhangfan_well@126.com。
基金资助:
国家转基因生物新品种培育重大专项(2009ZX08003-020B);国家肉鸡产业技术体系(nycytx-42-G2-05)资助。

Abstract

Abstract:

A xylanase gene xyn11G5 was cloned from Phialophora sp. G5 by degenerate PCR and TAIL-PCR method. Its ORF was 879 bp, encoded 292 amino acids residues, and the frontal 19 residues composed a putative signal peptide. Xyn11G5 was successfully expressed in Pichia pastoris and purified. The enzymatic characteristics analysis showed that the optimum pH and temperature for the recombinant xylanase was pH 5.0 and 50℃, respectively. It showed pH stability under neutral and thermo stability. Xyn11G5 was a good candidate in animal feed and bio-fuel industrial application.

Key words: Phialophora sp. G5, b-1, 4-xylanase, Pichia pastoris, enzyme characteristics

摘要：

通过简并PCR和TAIL-PCR技术从瓶霉Phialophora sp. G5菌株基因组DNA中克隆得到一个新的编码b-1,4-木聚糖酶的基因,命名为xyn11G5。该基因ORF全长879 bp,共编码292个氨基酸和一个终止密码子,前19个氨基酸为信号肽序列。将xyn11G5在毕赤酵母中进行分泌表达,重组蛋白经纯化达到电泳纯。酶学性质分析表明,该重组木聚糖酶最适温度为50℃,最适pH为5.0,在中性条件下具有良好的稳定性,并且具有一定的热稳定性,在饲料行业和生物能源行业领域具有潜在的应用前景。

关键词: Phialophora sp. G5;木聚糖酶;毕赤酵母;酶学性质

CLC Number:

ZHANG Fan1,2, SHI Peng-jun2, MIAO Li-hong1, YAO Bin2. Studies on Cloning of Acid Xylanase from Phialophora sp. G5 and its Characteristics[J]. , 2011, 13(3): 60-66.

张帆1,2,石鹏君2,缪礼鸿1,姚斌2. 来源于瓶霉Phialophora sp. G5的酸性木聚糖酶基因的克隆及性质的研究[J]. , 2011, 13(3): 60-66.

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Studies on Cloning of Acid Xylanase from Phialophora sp. G5 and its Characteristics

来源于瓶霉Phialophora sp. G5的酸性木聚糖酶基因的克隆及性质的研究

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