Cloning and expression of organic solvent tolerant lipase gene from Staphylococcus saprophyticus M36.
- Author:
Yanchong TANG
1
;
Yaping LU
;
Fengxia LÜ
;
Xiaomei BIE
;
Yao GUO
;
Zhaoxin LU
Author Information
1. Enzyme Engineering Laboratory, College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Base Sequence;
Cloning, Molecular;
Enzyme Stability;
Escherichia coli;
genetics;
metabolism;
Lipase;
biosynthesis;
genetics;
Molecular Sequence Data;
Organic Chemicals;
chemistry;
Recombinant Proteins;
biosynthesis;
genetics;
Solvents;
chemistry;
Staphylococcus saprophyticus;
enzymology
- From:
Chinese Journal of Biotechnology
2009;25(12):1989-1995
- CountryChina
- Language:Chinese
-
Abstract:
Lipases are important biocatalysts that are widely used in food processing and bio-diesel production. However, organic solvents could inactivate some lipases during applications. Therefore, the efficient cloning and expression of the organic solvent-tolerant lipase is important to its application. In this work, we first found out an organic solvent-tolerant lipase from Staphylococcus saprophyticus M36 and amplified the 741 bp Lipase gene lip3 (GenBank Accession No. FJ979867), by PCR, which encoded a 31.6 kD polypeptide of 247 amino acid residues. But the lipase shared 83% identity with tentative lip3 gene of Staphylococcus saprophyticus (GenBank Accession No. AP008934). We connected the gene with expression vector pET-DsbA, transformed it into Escherichia coli BL21 (DE3), and obtained the recombinant pET-DsbA-lip3. With the induction by 0.4 mmol/L of isopropyl beta-D-thiogalactopyranoside at pH 8.0, OD600 1.0, 25 degrees C for 12 h, the lipase activity reached up to 25.8 U/mL. The lipase expressed was stable in the presence of methanol, n-hexane, and isooctane, n-heptane.
