Purification, characterization, cloning and sequencing of phospholipase D from Streptomyces septatus TH-2

被引：60

作者：

Hatanaka, T ^{[1
]}

Negishi, T ^{[1
]}

Kubota-Akizawa, M ^{[1
]}

Hagishita, T ^{[1
]}

机构：

[1] Res Inst Biol Sci Okayama, Okayama 7161241, Japan

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 2002年 / 31卷 / 03期

关键词：

phospholipase D; Streptomyces; transphosphatidylation; purification;

D O I：

10.1016/S0141-0229(02)00121-7

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 [微生物学]; 0836 [生物工程]; 090102 [作物遗传育种]; 100705 [微生物与生化药学];

摘要：

Streptomyces septatus TH-2 produced 34 mg/l extracellular phospholipase D (PLD) when cultured in a medium containing glucose and citrate at 34degreesC for 4 days. The enzyme was purified to homogeneity by a one-column ion exchange procedure. PLD from S. septatus TH-2 gave K-m values of 0.99 and 0.67 mM for an artificial substrate, phosphatidyl-p-nitrophenol (PpNP), and k(cat) values of 86.7 and 258.8 s(-1) in hydrolytic and transphosphatidylation reactions, respectively. The PLD gene from S. septatus TH-2 was cloned and sequenced. Although the primary sequences of PLDs from S. septatus TH-2 and a commercially available PLD from Streptomyces sp. showed a high homology, these two PLDs showed similar kat values and different K-m values for PpNP and ethanol in transphosphatidylation. (C) 2002 Elsevier Science Inc. All rights reserved.

引用

页码：233 / 241

页数：9

共 32 条

[1]

Purification and applications of a phospholipase D from a new strain of Streptomyces [J].