Kinetics and mode of action of Aspergillus niger polygalacturonases

被引：22

作者：

Benen, JAE

Kester, HCM

Parenicova, L

Visser, J

机构：

[1] Section Molecular Genetics of Industrial Microorganisms, Wageningen Agricultural University, 6703 HA Wageningen

来源：

PECTINS AND PECTINASES | 1996年 / 14卷

关键词：

D O I：

10.1016/S0921-0423(96)80257-7

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 ; 0836 ; 090102 ; 100705 ;

摘要：

Endo-polygalacturonases I and II (PGI and PGII) isolated from recombinant A. niger were characterized with respect to pH optimum, activity on polygalacturonic acid (pga), mode of action and kinetics on oligogalacturonates. Vmax and Km values using pga as a substrate at the optimum pH 4.1 were calculated as 500 U/mg and 0.15 mg/ml and 2000 U/mg and 0.15 mg/ml for PGI and PGII, respectively. Mode of action analysis revealed a random cleavage pattern for PGII while for PGI multiple attack on a single chain was observed. For PGII a partial subsite map was obtained. Site directed mutagenesis of His223 of PGII with subsequent analysis of the mutated PGII revealed that His223 is essential for catalysis.

引用

页码：221 / 230

页数：10

共 15 条

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