ダウンロード数 : ?
ファイル
言語
英語
著者
ChoongSoo YUN Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University
松田 英幸
内容記述(抄録等)
Chitosanase (ChoA) from Mitsuaria chitosanitabida 3001 was successfully evolved with secretion efficiency and thermal stability. The inactive ChoA mutant (G151D) gene was used to mutate by an error-prone PCR technique and mutant genes that restored chitosanase activity were isolated. Two desirable mutants, designated M5S and M7T, were isolated. Two amino acids, Leu74 and Val75, in the signal peptide of ChoA were changed to Gln and Ile respectively in the M7T mutant, in addition to the G151D mutation. The L74Q/V75I double ChoA mutant was 1.5-fold higher in specific activity than wild-type ChoA due to efficient secretion of ChoA. One amino acid Asn222 was changed to Ser in the M5S mutant in addition to the G151D mutation. The N222S single ChoA mutant was 1.2-fold higher in specific activity and showed a 17% increase in thermal stability at 50 °C as compared with wild-type ChoA. This is the first study to achieve an evolutional increase in enzyme capability among chitosanses.
主題
chitosanase
random mutagenesis
error-prone PCR
thermostability
掲載誌名
Bioscience, Biotechnology, and Biochemistry
70
2
開始ページ
559
終了ページ
563
ISSN
0916-8451
ISSN(Online)
1347-6947
発行日
2006-01-01
DOI
出版者
Japan Society for Bioscience, Biotechnology, and Agrochemistry
資料タイプ
学術雑誌論文
ファイル形式
PDF
著者版/出版社版
著者版
部局
生物資源科学部
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