Increasing the thermal stability of the water-soluble pyrroloquinoline quinone glucose dehydrogenase by single amino acid replacement

被引：46

作者：

Sode, K ^{[1
]}

Ootera, T ^{[1
]}

Shirahane, M ^{[1
]}

Witarto, AB ^{[1
]}

Igarashi, S ^{[1
]}

Yoshida, H ^{[1
]}

机构：

[1] Tokyo Univ Agr & Technol, Dept Biotechnol, Tokyo 1848588, Japan

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 2000年 / 26卷 / 07期

关键词：

pyrroloquinoline quinone; glucose dehydrogenase; biosensor; thermal stability;

D O I：

10.1016/S0141-0229(99)00196-9

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：

Based on the characterization of a PCR mutation of water-soluble glucose dehydrogenase possessing pyrroloquinoline quinone (pQQ), PQQGDH-B, Ser231Cys, we have constructed a series of Ser231 variants. The replacement of Ser231 to Cys, Met, Leu, Asp, Asn, His, or Lys resulted in an increase in thermal stability. Among these variants, Ser231Lys showed the highest level of thermal stability and also showed high catalytic activity. Considering that Ser231Lys showed more than an 8-fold increase in its half-life during the thermal inactivation at 55 degrees C compared with the wild-type enzyme, and also retained catalytic activity similar to a wild-type enzyme, the application of this mutant enzyme as a glucose sensor constituent may develop into a stable glucose sensor construction. (C) 2000 Elsevier Science Inc. All rights reserved.

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页码：491 / 496

页数：6

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