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Applied and Environmental Microbiology, April 2003, p. 2383-2385, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.2383-2385.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Improving the Thermostability of Raw-Starch-Digesting Amylase from a Cytophaga sp. by Site-Directed Mutagenesis

Rong-Jen Shiau, Hui-Chen Hung, and Chii-Ling Jeang*

Department of Food Science, National Chung Hsing University, Taichung, Taiwan, Republic of China

Received 23 September 2002/ Accepted 15 January 2003

A heat-stable raw-starch-digesting amylase (RSDA) was generated through PCR-based site-directed mutagenesis. At 65°C, the half-life of this mutant RSDA, which, compared with the wild-type RSDA, lacks amino acids R178 and G179, was increased 20-fold. While the wild type was inactivated completely at pH 3.0, the mutant RSDA still retained 41% of its enzymatic activity. The enhancement of RSDA thermostability was demonstrated to be via a Ca2+-independent mechanism.

* Corresponding author. Mailing address: Department of Food Science, National Chung Hsing University, 250 Kuokang Rd., Taichung 402, Taiwan, Republic of China. Phone and fax: 886-422862797. E-mail: cljeang{at}nchu.edu.tw.

Applied and Environmental Microbiology, April 2003, p. 2383-2385, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.2383-2385.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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