Purification and characterization of solvent tolerant lipase from Bacillus sphaericus MTCC 7542

被引：15
作者：
Tamilarasan, K. [1 ]
Kumar, M. Dharmendira [2 ]
机构：
[1] Madha Engn Coll, Dept Biotechnol, Chennai 600069, Tamil Nadu, India
[2] Anna Univ, Dept Appl Sci & Technol, Chennai 600025, Tamil Nadu, India
来源：
BIOCATALYSIS AND AGRICULTURAL BIOTECHNOLOGY | 2012年 / 1卷 / 04期
关键词：
Molecular weight; Zymogram; Electrophoresis; Stability; Lipase activity;
D O I：
10.1016/j.bcab.2012.07.001
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：
Solvent tolerant extracellular lipase from Bacillus sphaericus MTCC 7542 was purified by DEAESepharose anion exchange chromatography. This purification resulted in 377 U/mg specific activity and 17.33-fold with 5.7% recovery obtained. The molecular weight of the purified lipase was determined to be 69 kDa using SDS polyacrylamide gel electrophoresis and conformed by zymogram. The purified lipase showed optimal activity and stability at pH 8.0 and 40 degrees C. The purified lipase was stable in the presence of hydrophobic solvents like n-butanol, benzene, hexane and toluene. Lipase activity was promoted in the presence of Mg2+, Ca2+, Cu2+, K+ and Tween 80. (C) 2012 Elsevier Ltd. All rights reserved.
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收藏
页码：309 / 313
页数：5
相关论文
共 21 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]   DISC ELECTROPHORESIS .2. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS [J].
DAVIS, BJ .
ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1964, 121 (A2) :404-&
[3]   Biochemical analysis of a native and proteolytic fragment of a high-molecular-weight thermostable lipase from a mesophilic Bacillus sp. [J].
Dosanjh, NS ;
Kaur, J .
PROTEIN EXPRESSION AND PURIFICATION, 2002, 24 (01) :71-75
[4]   Molecular cloning and characterization of two thermostable carboxyl esterases from Geobacillus stearothermophilus [J].
Ewis, HE ;
Abdelal, AT ;
Lu, CD .
GENE, 2004, 329 :187-195
[5]  
Gupta J.K., 2000, J PUNJAB ACAD SCI, V2, P75
[6]   Bacterial lipases: an overview of production, purification and biochemical properties [J].
Gupta, R ;
Gupta, N ;
Rathi, P .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2004, 64 (06) :763-781
[7]   Industrial applications of microbial lipases [J].
Hasan, Fariha ;
Shah, Aamer Ali ;
Hameed, Abdul .
ENZYME AND MICROBIAL TECHNOLOGY, 2006, 39 (02) :235-251
[8]   Bacterial biocatalysts: Molecular biology, three-dimensional structures, and biotechnological applications of lipases [J].
Jaeger, KE ;
Dijkstra, BW ;
Reetz, MT .
ANNUAL REVIEW OF MICROBIOLOGY, 1999, 53 :315-+
[9]   Lipases for biotechnology [J].
Jaeger, KE ;
Eggert, T .
CURRENT OPINION IN BIOTECHNOLOGY, 2002, 13 (04) :390-397
[10]   Directed evolution and the creation of enantioselective biocatalysts [J].
Jaeger, KE ;
Eggert, T ;
Eipper, A ;
Reetz, MT .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2001, 55 (05) :519-530
123