Cloning and expression of a lipase gene from Bacillus subtilis FS1403 in Escherichia coli

被引：8

作者：

Shi, Bihong ^{[1
,2
]}

Wu, Weibin ^{[1
]}

Wen, Jianxin ^{[1
]}

Shi, Qiaoqin ^{[2
]}

Wu, Songgang ^{[2
]}

机构：

[1] Min Hou Univ Town, Fujian Normal Univ, Coll Life Sci, Fuzhou 350108, Peoples R China

[2] Fujian Normal Univ, Engn Res Ctr Ind Microbiol, Minist Educ, Fuzhou 350108, Peoples R China

来源：

ANNALS OF MICROBIOLOGY | 2010年 / 60卷 / 03期

关键词：

Bacillus subtilis; Lipase; Gene cloning; Expression; NUCLEOTIDE-SEQUENCE; THERMOSTABLE LIPASE; MOLECULAR-CLONING; PURIFICATION;

D O I：

10.1007/s13213-010-0055-y

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A moderate heat-resistant lipase gene from Bacillus subtilis FS1403 was cloned. Sequence analysis revealed that the lipase gene encodes a 212-amino-acid protein containing the conserved motif Ala-X-Ser-X-Gly that is present in other Bacillus lipases. Phylogenetic analysis suggested that the B. subtilis FS1403 lipase gene belongs to family I. 4 of bacterial lipase genes. The lipase gene was inserted in pET-28a(+) and expressed in Escherichia coli BL21. SDS-PAGE analysis revealed that the recombinant lipase had a molecular weight of approximately 23 kDa, which corresponded to the predicted size (22.8 kDa) of the deduced protein. The lipase activity was the highest at 50 degrees C and at pH 8.0. The enzyme retained over 70% of the maximal activity at temperatures of 40-60 degrees C and pH 6.0-11.0.

引用

页码：399 / 404

页数：6

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