Purification and Characterization of Organic Solvent and Detergent Tolerant Lipase from Thermotolerant Bacillus sp RN2

被引：32

作者：

Kanjanavas, Pornpimon ^{[1
]}

Khuchareontaworn, Sintawee ^{[1
]}

Khawsak, Paisarn ^{[1
]}

Pakpitcharoen, Arda ^{[1
]}

Pothivejkul, Khajeenart ^{[2
]}

Santiwatanakul, Somchai ^{[3
]}

Matsui, Kenji ^{[4
]}

Kajiwara, Tadahiko ^{[4
]}

Chansiri, Kosum ^{[1
]}

机构：

[1] Srinakharinwirot Univ, Fac Med, Dept Biochem, Bangkok 10110, Thailand

[2] Srinakharinwirot Univ, Fac Sci, Dept Biol, Bangkok 10110, Thailand

[3] Srinakharinwirot Univ, Fac Med, Dept Pathol, Bangkok 10110, Thailand

[4] Yamaguchi Univ, Fac Agr, Dept Biol Chem, Yamaguchi 753, Japan

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2010年 / 11卷 / 10期

关键词：

lipase; organic tolerant; detergent tolerant; Bacillus; thermotolerant; BIOTECHNOLOGICAL APPLICATIONS; THERMOSTABLE LIPASE; PSEUDOMONAS-CEPACIA; GENE; LICHENIFORMIS; SEQUENCE; EXPRESSION; PROTEINS; CLONING; ENZYME;

D O I：

10.3390/ijms11103783

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The aim of this study was to characterize the organic solvent and detergent tolerant properties of recombinant lipase isolated from thermotolerant Bacillus sp. RN2 (Lip-SBRN2). The isolation of the lipase-coding gene was achieved by the use of inverse and direct PCR. The complete DNA sequencing of the gene revealed that the lip-SBRN2 gene contains 576 nucleotides which corresponded to 192 deduced amino acids. The purified enzyme was homogeneous with the estimated molecular mass of 19 kDa as determined by SDS-PAGE and gel filtration. The Lip-SBRN2 was stable in a pH range of 9-11 and temperature range of 45-60 degrees C. The enzyme was a non metallo-monomeric protein and was active against pNP-caprylate (C8) and pNP-laurate (C12) and coconut oil. The Lip-SBRN2 exhibited a high level of activity in the presence of 108% benzene, 102.4% diethylether and 112% SDS. It is anticipated that the organic solvent and detergent tolerant enzyme secreted by Bacillus sp. RN2 will be applicable as catalysts for reaction in the presence of organic solvents and detergents.

引用

页码：3783 / 3792

页数：10

共 23 条

[1]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[2] MINIREVIEW ON PANCREATIC LIPASE AND COLIPASE [J].

CHAPUS, C ;

ROVERY, M ;

SARDA, L ;

VERGER, R .

BIOCHIMIE, 1988, 70 (09) :1223-1234

[3] The alpha/beta fold family of proteins database and the cholinesterase gene server ESTHER [J].

Cousin, X ;

Hotelier, T ;

Giles, K ;

Lievin, P ;

Toutant, JP ;

Chatonnet, A .

NUCLEIC ACIDS RESEARCH, 1997, 25 (01) :143-146

[4] Application of the partial 16S rDNA sequence as an index for rapid identification of species in the genus Bacillus [J].

Goto, K ;

Omura, T ;

Hara, Y ;

Sadaie, Y .

JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY, 2000, 46 (01) :1-8

[5] Lichenysin -: A more efficient cation chelator than surfactin [J].

Grangemard, I ;

Wallach, J ;

Maget-Dana, R ;

Peypoux, F .

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 2001, 90 (03) :199-210

[6] A new tannase substrate for spectrophotometric assay [J].

Iacazio, G ;

Périssol, C ;

Faure, B .

JOURNAL OF MICROBIOLOGICAL METHODS, 2000, 42 (03) :209-214

[7] 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-+

[8] BIOSURFACTANTS FROM BACILLUS-LICHENIFORMIS - STRUCTURAL-ANALYSIS AND CHARACTERIZATION [J].

JENNY, K ;

KAPPELI, O ;

FIECHTER, A .

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1991, 36 (01) :5-13

[9] Gene cloning and characterization of thermostable lipase from Bacillus stearothermophilus L1 [J].

Kim, HK ;

Park, SY ;

Lee, JK ;

Oh, TK .

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1998, 62 (01) :66-71

[10] A SIMPLE AND RAPID COLORIMETRIC METHOD FOR DETERMINATION OF FREE FATTY-ACIDS FOR LIPASE ASSAY [J].

KWON, DY ;

RHEE, JS .

JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY, 1986, 63 (01) :89-92

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