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language
eng
Author
ChoongSoo YUN Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University
Matsuda, Hideyuki
Description
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.
Subject
chitosanase
random mutagenesis
error-prone PCR
thermostability
Journal Title
Bioscience, Biotechnology, and Biochemistry
Volume
70
Issue
2
Start Page
559
End Page
563
ISSN
0916-8451
ISSN(Online)
1347-6947
Published Date
2006-01-01
DOI
Publisher
Japan Society for Bioscience, Biotechnology, and Agrochemistry
NII Type
Journal Article
Format
PDF
Text Version
著者版
OAI-PMH Set
Faculty of Life and Environmental Science