一种来源于Paenibacillus sp. A1的中性β-甘露
聚糖酶基因克隆及其酶学性质研究

›› 2009, Vol. 11 ›› Issue (5): 60-65.

一种来源于Paenibacillus sp. A1的中性β-甘露
聚糖酶基因克隆及其酶学性质研究

刘小丹1,2,杨培龙2,刘永超3,许修宏1,孟昆2,姚斌2

(1.东北农业大学资源与环境学院, 哈尔滨150030|2.中国农业科学院饲料研究所, 北京 100081|
3.山东省莒南县农业局, 山东莒南 276600）

收稿日期:2009-03-24 修回日期:2009-07-08 出版日期:2009-10-15 发布日期:2009-10-15
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:010-82106063;E-mail:yaobin@mail.caas.net.cn。许修宏,教授,博士,博士生导师,从事应用微生物研究。
作者简介:刘小丹,硕士研究生,研究方向为微生物工程。
基金资助:
农业微生物菌种资源整理整合及共享项目（2005DKA21201）;国家863计划项目（2006AA02Z213）资助。

Studies on the Gene Cloning and its Enzymology Characteristics of a Neutral β-Mannanase from Paenibacillus sp.A1

LIU Xiao-dan1,2, YANG Pei-long2, LIU Yong-chao3, XU Xiu-hong2, |MENG Kun2, YAO Bin2

(1.School of Resources &|Environment, Northeast Agriculture University, Harbin 150030|2.Feed Research Institute,
Chinese Academy of Agricultural Sciences, Beijing 100081|3.Agriculture Bureau of Junan County, |Shandong |Junan 276600, China)

Received:2009-03-24 Revised:2009-07-08 Online:2009-10-15 Published:2009-10-15

摘要/Abstract

摘要：

采用简并PCR和TAIL-PCR技术,从来源于天山冰川土壤的类芽孢杆菌Paenibacillus sp. A1菌株基因组中扩增出β-甘露聚糖酶基因编码序列。序列分析表明,该基因全长960 bp,编码一个新的糖苷水解酶第5家族的β-甘露聚糖酶。将该基因克隆到原核表达载pET-22b(+)中,经过IPTG诱导表达,酶的胞外表达量达0.62 U/mL。酶学特性分析表明其最适pH为6.5,最适温度为55℃,属中性β-甘露聚糖酶,具有较好的底物适应性,适宜用于鱼类等水产动物养殖饲料行业中。

关键词: β-甘露聚糖酶, Paenibacillus sp. A1;基因克隆;表达;酶学性质

Abstract:

A β-mannanase encoding gene was isolated from strain of Paenibacillus sp.A1 by degenerate PCR and TAIL-PCR. Sequence analysis shows that the gene of 960 bp length encodes a novel β-mannanase, which belongs to glycosyl hydrolase family 5. The gene was ligated into the prokaryotic expression vector pET-22b(+), then expressed with the enzyme activity of 0.62 U/mL after induction by IPTG. Enzymatic characteristics analysis showed that the optimum pH for the enzyme was 6.5 and the optimum temperature was 55℃. It is a neutral β-mannanase, which has better substrate adaptability. It is suitable to be taken as feed additives for fish and other aquaculture industry.

Key words: β-mannanase, Paenibacillus sp. A1, gene cloning, expression, enzymology characteristics

中图分类号:

Q556+.2

刘小丹1,2,杨培龙2,刘永超3,许修宏1,孟昆2,姚斌2. 一种来源于Paenibacillus sp. A1的中性β-甘露
聚糖酶基因克隆及其酶学性质研究[J]. , 2009, 11(5): 60-65.

LIU Xiao-dan1,2, YANG Pei-long2, LIU Yong-chao3, XU Xiu-hong2, . Studies on the Gene Cloning and its Enzymology Characteristics of a Neutral β-Mannanase from Paenibacillus sp.A1[J]. , 2009, 11(5): 60-65.