An in vitro evaluation of a thermostable pectate lyase by using error-prone PCR

被引：2

作者：

Nakaniwa, T ^{[1
]}

Tada, T

Takao, M

Sakai, T

Nishimura, K

机构：

[1] Osaka Prefecture Univ, Adv Sci & Technol Res Inst, Sakai, Osaka 5998570, Japan

[2] IGA BioRes Co Ltd, Amagasaki, Hyogo 6600806, Japan

来源：

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC | 2004年 / 27卷 / 2-3期

关键词：

pectate lyase; thermostability; error-prone PCR; random mutation;

D O I：

10.1016/j.molcatb.2003.10.005

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：

Thermostable pectate lyase 47 (PL 47) prepared from Bacillus sp. TS 47 was mutated by a random mutation with error-prone PCR. The mutated PL 47 gene was expressed in Escherichia coli and the variants were screened over the PL activity using a plate assay method. Three less-thermostable mutants were isolated and their enzymes (LT-PLs) were isolated and characterized. The LT-PLs were inferior to the PL 47 in their thermostability and thermoactivity. Analyses of the amino acids substituted in the LT-PLs suggested that Ala325 in the PL 47 contributes to its thermostability. (C) 2003 Elsevier B.V. All rights reserved.

引用

页码：127 / 131

页数：5

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