地衣芽孢杆菌W10 β-甘露聚糖酶基因的克隆与表达  

Cloning and expression of β-mannanase gene from Bacillus licheniformis W10

作　　者：张清霞[1] 黄姗姗[1] 童蕴慧[1] 徐敬友[1] 

机构地区：[1]扬州大学园艺与植物保护学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2010年第4期77-81,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(30571243);江苏省高校自然科学基金资助项目(09KJB210005)

摘　　要：地衣芽孢杆菌W10菌株具有较高的β-甘露聚糖酶活性。采用PCR技术从W10基因组中扩增出β-甘露聚糖酶基因序列,通过克隆、测序获得1 017 bp甘露聚糖酶基因manW10,其编码338个氨基酸。以pET-22b(+)为载体和pelB为信号肽,将manW10转入EscherichiacoliBL21(DE3)中进行表达,经IPTG诱导酶活性达19.73 U.mL-1。经镍柱亲和层析获得较纯的β-甘露聚糖酶,其分子质量约为38 ku。The high β-mannanase activity was screened in Bacillus licheniformis W10.The β-mannanase gene manW10 was cloned from W10 genome by using PCR and consisted of a 1 017 bp sequence encoding 338 amino acids.The gene was constructed in an expression vector pET22b（＋） with pelB singal peptide,transformed and expressed in Escherichia coli BL21（DE3）.The β-mannanase activity expressed in vitro was 19.73 U·mL-1 under the induction of IPTG.The enzyme was purified by affinity chromatography of nikel and had a molecular weight of approximately 38 ku.

关 键 词：地衣芽孢杆菌 Β-甘露聚糖酶 基因克隆 表达 

分 类 号：Q785[生物学—分子生物学] S154.38[农业科学—土壤学]