Gene Cloning, High-Level Expression, and Characterization of an Alkaline and Thermostable Lipase from Trichosporon coremiiforme V3

被引：5
作者：
Wang, Jian-Rong [1 ,2 ]
Li, Yang-Yuan [1 ,2 ]
Liu, Danni [1 ,2 ]
机构：
[1] Guangdong VTR Biotech Co Ltd, Zhuhai 519060, Guangdong, Peoples R China
[2] Guangdong Feed Addit Res & Biotechnol Ctr, Zhuhai 519060, Guangdong, Peoples R China
来源：
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY | 2015年 / 25卷 / 06期
关键词：
Trichosporon coremiiforme; alkaline and thermostable lipase; high-level expression; Pichia pastoris; ORGANIC-SOLVENT-TOLERANT; PICHIA-PASTORIS; CANDIDA-RUGOSA; EXTRACELLULAR LIPASE; PURIFICATION; OPTIMIZATION;
D O I：
10.4014/jmb.1408.08039
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：
The present study describes the gene cloning and high-level expression of an alkaline and thermostable lipase gene from Trichosporon coremiiforme V3. Nucleotide analysis revealed that this lipase gene has an open reading frame of 1,692 bp without any introns, encoding a protein of 563 amino acid residues. The lipase gene without its signal sequence was cloned into plasmid pPICZ alpha A and overexpressed in Pichia pastoris X33. The maximum lipase activity of recombinant lipase was 5,000 U/ml, which was obtained in fed-batch cultivation after 168 h induction with methanol in a 50 L bioreactor. The purified lipase showed high temperature tolerance, and being stable at 60 degrees C and kept 45% enzyme activity after 1 h incubation at 70 degrees C. The stability, effects of metal ions and other reagents were also determined. The chain length specificity of the recombinant lipase showed high activity toward triolein (C18:1) and tripalmitin (C16:0).
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页码：845 / 855
页数：11
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