文献类型： 外文期刊

作者： Fu, Jianhong ¹ ; Huang, Huoqing ³ ; Meng, Kun ³ ; Yuan, Tiezheng ³ ; Yao, Bin ³ ; Shi, Yuhu ¹ ; Ouyang, Pingkai ¹ ;

作者机构： 1.Nanjing Technol Univ, Nanjing 210009, Peoples R China

2.Xinjiang Acad Agr Sci, Inst Appl Microbiol, Urumqi 830000, Peoples R China

3.Chinese Acad Agr Sci, Feed Res Inst, Beijing 100081, Peoples R China

关键词： cold-adapted phospholipase A(1);Serratia sp.;expression;characterization

期刊名称：ENZYME AND MICROBIAL TECHNOLOGY （ 影响因子：3.493； 五年影响因子：3.699 ）

ISSN： 0141-0229

年卷期： 2008 年 42 卷 2 期

页码：

收录情况： SCI

摘要： The gene encoding a cold-adapted phospholipase A(1) (PLA(1)) from a psychrotrophic, glacier soil bacterium Serratia sp. xjF1 was cloned by two-step PCR (general PCR and TAIL-PCR). The full-length fragment comprised two open reading frames plA and plS. The gene product of plA encoding 320 amino acids with a molecular weight of 33.8 kDa was identified as a phospholipase A(1). Its amino acid sequence exhibited the highest homology to PLA(1) of Serratia marcescens (71%). plS encoded a protein of 251 amino acids, which showed no enzymatic activity. The result of plA expression in Escherichia coli indicated that plS might improve the efficient expression of PLA(1) in E coli. Furthermore, PLA(1) was functionally expressed in Pichia pastoris, yielding 41.8 U/mL in a 3.7L fermentor. The purified recombinant phospholipase A(1) (rPLA(1)) had features typical of cold-adapted enzymes with a temperature optimum of 35 degrees C and a maximum activity of 70% at 10 degrees C. The rate of catalysis was optimal at pH 9.0 and the enzyme could be slightly activated by Ca2+. This is the first report on gene isolation and expression of cold-adapted PLA(1). (C) 2007 Elsevier Inc. All rights reserved.