Enhancement of the thermostability of the maltogenic amylase MAUS149 by Gly312Ala and Lys436Arg substitutions

被引：34

作者：

Ben Mabrouk, Sameh ^{[1
]}

Aghajari, Nushin ^{[2
]}

Ben Ali, Mamdouh ^{[1
]}

Ben Messaoud, Ezzedine ^{[1
]}

Juy, Michel ^{[2
]}

Haser, Richard ^{[2
]}

Bejar, Samir ^{[1
]}

机构：

[1] Ctr Biotechnol Sfax, Lab Microorganismes & Biomol, Sfax 3018, Tunisia

[2] UMR 5086 CNRS Univ Lyon 1, Lab BioCristallog, Inst Biol & Chim Prot, BioSci Gerland Lyon Sud IFR128, F-69367 Lyon 07, France

来源：

BIORESOURCE TECHNOLOGY | 2011年 / 102卷 / 02期

关键词：

Maltogenic amylase; Site-directed mutagenesis; Thermostability; Salt bridge; Hydrophobic interactions; ALPHA-AMYLASE; THERMAL-STABILITY; SEQUENCE; ENZYMES; NEOPULLULANASE; PROTEINS; ACARBOSE; BINDING; FAMILY; CYCLOMALTODEXTRINASE;

D O I：

10.1016/j.biortech.2010.08.082

中图分类号：

S2 [农业工程];

学科分类号：

0828 ;

摘要：

Based on sequence alignments and homology modeling, Gly 312 and Lys 436 of the maltogenic amylase from Bacillus sp. US149 (MAUS149) were selected as targets for site-directed mutagenesis to improve the thermostability of the enzyme. Variants of MAUS149 with amino acid substitutions G312A, K436R and G312A-K436R had substrate specificities, kinetic parameters and pH optima similar to those of the wild-type enzyme: however, the enzymes with substitutions K436R and G312A-K436R, had an optimal temperature of 45 degrees C instead of the 40 degrees C for the wild-type enzyme. The half-life time at 55 degrees C increased from 15 to 25 min for the double mutant. Molecular modeling suggests that the increase in thermostability was due to new hydrophobic interactions and the formation of a salt bridge and hydrogen bond in the G312A and K436R variants, respectively. The double mutant could be a potential candidate for application in the bread industry. (C) 2010 Elsevier Ltd. All rights reserved.

引用

页码：1740 / 1746

页数：7

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