Improvement of the Thermal Stability of a Calcium-free, Alkaline .ALPHA.-Amylase by Site-directed Mutagenesis
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- Ozawa Tadahiro
- Tochigi Research Laboratories, Kao Corporation
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- Endo Keiji
- Tochigi Research Laboratories, Kao Corporation
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- Igarashi Kazuaki
- Tochigi Research Laboratories, Kao Corporation
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- Kitayama Kaori
- Tochigi Research Laboratories, Kao Corporation
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- Hayashi Yasuhiro
- Tochigi Research Laboratories, Kao Corporation
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- Hagihara Hiroshi
- Tochigi Research Laboratories, Kao Corporation
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- Kawai Shuji
- Tochigi Research Laboratories, Kao Corporation
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- Ito Susumu
- Creative Research Initiative “Sousei”(CRIS), Hokkaido University
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- Ozaki Katsuya
- Tochigi Research Laboratories, Kao Corporation
Bibliographic Information
- Other Title
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- 部位特異的変異によるカルシウム非依存性アルカリα-アミラーゼの熱安定性の向上
- Improvement of the thermal stability of a calcium-free, alkaline α-amylase by site-directed mutagenesis
- Improvement of the thermal stability of a calcium free alkaline a amylase by site directed mutagenesis
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Abstract
Alkaline α-amylase from Bacillus sp. strain KSM-K38 (AmyK38) is a calcium-free enzyme that is stable against chelating and oxidative reagents. Recently, the thermostability of this enzyme was improved by replacement of its amino-terminal 11 amino acid residues with the corresponding residues of α-amylase from Bacillus sp. strain KSM-1378. In this study, to further improve the thermal stability, we compared the three-dimensional structure of AmyK38 with that of a hyper-thermostable α-amylase from Bacillus licheniformis. Using site-directed mutagenesis, we created a new possible ionic interaction in the flexible loop region from Gln167 to Gln170 of AmyK38 because the corresponding region in α-amylase from B. licheniformis has an ionic interaction between Glu167 and Lys170. Substitution of Gln167 or Tyr169 with Glu or Lys, respectively, was found to enhance the thermostability of AmyK38. Combination of both substitutions with replacement of the amino-terminal 11 residues further improved the thermostability of the enzyme.
Journal
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- Journal of Applied Glycoscience
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Journal of Applied Glycoscience 54 (2), 77-83, 2007
The Japanese Society of Applied Glycoscience
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Details 詳細情報について
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- CRID
- 1390282681270456064
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- NII Article ID
- 10018617400
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- NII Book ID
- AN10453916
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- ISSN
- 18807291
- 13447882
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- NDL BIB ID
- 8827830
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed