Improvement of heat-stability of subtilisin J by protein-engineering = 단백질 공학을 이용한 서브틸리신 J의 열안정성 증가

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dc.contributor.advisorByun, Si-Myung-
dc.contributor.advisor변시명-
dc.contributor.authorBae, Kwang-Hee-
dc.contributor.author배광희-
dc.date.accessioned2011-12-12T08:59:43Z-
dc.date.available2011-12-12T08:59:43Z-
dc.date.issued1994-
dc.identifier.urihttp://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=69126&flag=dissertation-
dc.identifier.urihttp://hdl.handle.net/10203/28419-
dc.description학위논문(석사) - 한국과학기술원 : 생명과학과, 1994.2, [ vi, 49 p. ]-
dc.description.abstractSer-49 in subtilisin J, an extracellular serine protease from B. stearothermophilus, is highly conserved residue among subtilisins from Bacillus sp. Moreover, Ala-48 - Ser-49 peptide bond was known as a primary autolysis site in subtilisin BPN`` recently. Previously constructed Asp-49 mutant protein, corresponding to that a more thermostable subtilisin variants, shows proteolytic activity, but so unstable that it was degraded during cultivation in Tyr-58 - Gln-59 peptide bond. The enzymatic activity of the Asp-49 mutant protein is higher than that of wild-type in early growth phase. To elucidate the role of Ser-49 and increase the heat-stability, first, the Ser-49 was replaced with Thr,Asn and Glu. From results of substitution of Ser-49, we concluded that hydroxyl of Ser-49, as H-bond donor and acceptor, is important in prevention of autolysis. Second, in consideration of substrate specificity of subtilisin, The Tyr-58 or Gln-59 weae replaced with various amino acid using Asp-49 mutant as a template. Among the mutants, Asp-49 -$\Delta$Tyr-58 mutant showed a considerably improved resistance to heat. These results demonstrate the powerful probability of protein-engineering in enhancement of heat stability of various enzymes.eng
dc.languageeng-
dc.publisher한국과학기술원-
dc.subject부위 특이 돌연변이 유발.-
dc.titleImprovement of heat-stability of subtilisin J by protein-engineering = 단백질 공학을 이용한 서브틸리신 J의 열안정성 증가-
dc.typeThesis(Master)-
dc.identifier.CNRN69126/325007-
dc.description.department한국과학기술원 : 생명과학과, -
dc.identifier.uid000923217-
dc.contributor.localauthorByun, Si-Myung-
dc.contributor.localauthor변시명-
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BS-Theses_Master(석사논문)
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