Cloning, heterologous expression, and enzymatic characterization of a thermostable glucoamylase from Talaromyces emersonii

被引：26

作者：

Nielsen, BR ^{[1
]}

Lehmbeck, J ^{[1
]}

Frandsen, TP ^{[1
]}

机构：

[1] Novozymes AS, DK-2880 Bagsvaerd, Denmark

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2002年 / 26卷 / 01期

关键词：

thermostability; specific activity; saccharifications; reversion; cloning;

D O I：

10.1016/S1046-5928(02)00505-3

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The gene encoding a thermostable glucoamylase from Talaromyces emersonii was cloned and, subsequently, heterologously expressed in Aspergillus niger. This glucoamylase gene encodes a 618 amino acid long protein with a calculated molecular weight of 62,827 Da. T. emersonii glucoamylase fall into glucoside hydrolase family 15, showing approximately 60%, sequence similarity to glucoamylase from A. niger. The expressed enzyme shows high specific activity towards maltose, isomaltose, and maltoheptaose, having 3-6-fold elevated k(cat) compared to A. niger glucoamylase. T emersonii glucoamylase showed significantly improved thermostability with a half life of 48 h at 65 degrees C in 30% (w/v) glucose, compared to 10 h for glucoamylase from A. niger. The ability of the glucoamylase to hydrolyse amylopectin at 65 degrees C is improved compared to A. niger glucoamylase, giving a significant higher final glucose yield at elevated temperatures. The increased thermal stability is thus reflected in the industrial performance, allowing T emersonii glucoamylase to operate at a temperature higher than the A. niger enzyme. (C) 2002 Elsevier Science (USA). All rights reserved.

引用

页码：1 / 8

页数：8

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