Enhanced thermal stability and catalytic efficiency of an alkaline protease from pseudomonas sp. KFCC10818 by changing the autolysis sitesPseudomonas sp. KFCC10818로부터 분리된 염기성단백질분해효소의 자가 분해성을 방해함으로써 증진된 열안정성 및 촉매능

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dc.contributor.advisorYoo, Ook-Joon-
dc.contributor.advisor유욱준-
dc.contributor.authorJang, Jae-Woo-
dc.contributor.author장재우-
dc.date.accessioned2011-12-12T09:01:54Z-
dc.date.available2011-12-12T09:01:54Z-
dc.date.issued1998-
dc.identifier.urihttp://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=135371&flag=dissertation-
dc.identifier.urihttp://hdl.handle.net/10203/28564-
dc.description학위논문(석사) - 한국과학기술원 : 생물과학과, 1998.2, [ viii, 55 p. ]-
dc.description.abstractThe extracellular alkaline protease, AprP, isolated from Pseudomonas sp. KFCC10818 was degraded by autoproteolysis at high temperature. Two sites of autoproteolysis were identified as amino terminal of Ser307 and Ser331 by isolation of early autolysis products and amino terminal sequence analysis. These sites seemed to be located in an exposed surface loop by fitting the sequence to model for subtilisin BPN``. To enhance thermal stability of AprP, we replaced serine residues of these sites with aspartate residues by site-directed mutagenesis. However, the only S331D mutant of two mutants was successfully purified and characterized. This mutant enzyme had a Km value 2.4-fold and a Kcat value 2-fold smaller than wild-type as measured by hydrolysis of s-AAPF-pna at pH 10.5. The half-life of S331D mutant showed 1.5 and 2.4 times longer than wild-type at 55℃ and 60℃, respectively. Therefore, these results suggest that the change of autolytic cleavage sites enhance the thermal stability and catalytic efficiency of the enyme AprP by preventing from cleaving.eng
dc.languageeng-
dc.publisher한국과학기술원-
dc.subjectThermal stability-
dc.subjectAutolysis-
dc.subjectAlkaline protease-
dc.subjectSite-directed mutagenesis-
dc.subject특정부위 돌연변이-
dc.subject열안정-
dc.subject자가분해성-
dc.subject염기성단백질-
dc.titleEnhanced thermal stability and catalytic efficiency of an alkaline protease from pseudomonas sp. KFCC10818 by changing the autolysis sites-
dc.title.alternativePseudomonas sp. KFCC10818로부터 분리된 염기성단백질분해효소의 자가 분해성을 방해함으로써 증진된 열안정성 및 촉매능-
dc.typeThesis(Master)-
dc.identifier.CNRN135371/325007-
dc.description.department한국과학기술원 : 생물과학과, -
dc.identifier.uid000963538-
dc.contributor.localauthorYoo, Ook-Joon-
dc.contributor.localauthor유욱준-
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