Enhancement of the thermostability and the catalytic efficiency of an alkaline protease from pseudomonas sp. KFCC10818 by site-directed mutagenesis = Pseudomonas sp. KFCC 10181로 부터 분리된 염기성 단백질분해효소의 열안정성 및 촉매능의 중진

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To obtain increasement of industrial value by enhancement of thermostability, site-directed mutagenesis in an alkaline protease, AprP from Pseudomonas sp. KFCC 10818 was carried out. By sequence alignments with aqualysin I of Thermus aquaticus YT-1, sites for cysteine substitutions to form a disulfide bond in this protease, AprP, were chosen. Gly199 and Phe236 were replaced by cysteine residues in the enzyme by site-directed mutagenesis. The G199C/F236C mutant enzyme appeared to form a disulfide bond spontaneously in its expression and showed improved kinetic parameters compared to that of the wild-type enzyme for hydrolysis of an artificial synthetic substrate at pH 8.5 and pH 10.5. The half-life of the G199C/F236C mutant was found to be 2 - 4.8 times longer than that of the wild-type enzyme depending on experimental conditions. There was not any significant differences in the half-life of both enzymes when examined under reducing condition. These results suggest that the introduction of a disulfide bond by site-directed mutagenesis enhanced both of the thermostability and the catalytic efficiency of the enzyme.
Advisors
유욱준researcherYoo, Ook-Joonresearcher
Description
한국과학기술원 : 생물과학과,
Publisher
한국과학기술원
Issue Date
1996
Identifier
115828/325007 / 000943016
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생물과학과, 1996.2, [ viii, 75 p. ]

Keywords

Alkaline protease; Thermostability; 열안정성; 염기성 단백질분해효소; Site-directed mutagenesis

URI
http://hdl.handle.net/10203/28537
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=115828&flag=dissertation
Appears in Collection
BS-Theses_Master(석사논문)
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