In vitro formation of active carboxypeptidase Y from pro-carboxypeptidase Y inclusion bodies by fed-batch operation

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dc.contributor.authorMoon Sun Hahm-
dc.contributor.authorBong Hyun Chung-
dc.date.accessioned2017-04-19T08:58:29Z-
dc.date.available2017-04-19T08:58:29Z-
dc.date.issued2001-
dc.identifier.issn1017-7825-
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/5621-
dc.description.abstractThe gene encoding yeast pro-carboxypeptidase Y (pro-CPY) has been cloned and expressed in Escherichia coli. Most of the expressed pro-CPY was accumulated as cytoplasmic insoluble aggregates. In our previous study [3], active CPY was obtained by renaturation of entirely denatured pro-CPY followed by in vitro proteolytic processing with proteinase K along with the activation process. The same refolding process was performed to produce an active CPY from pro-CPY inclusion bodies with renaturation buffers containing proteinase K at different concentrations. The refolding efficiency decreased from 25% to 2% in the renaturation buffers containing proteinase K at concentrations of 60 μg/ml and 0.6 μg/ml, respectively. In an attempt to increase the refolding efficiency with a lesser amount of proteinase K, a novel fed-batch refolding process was developed. In a fed-batch refolding, 99 ml of the renaturation buffer containing pro-CPY was gradually added into 1 ml of the renaturation buffer containing 60 μg/ml of proteinase K to give a final proteinase K concentration of 0.6 μg/ml. The fed-batch refolding process resulted in a refolding efficiency of 18%, which corresponded to a 9-fold increase over that (2%) in the batch process.-
dc.publisherKorea Soc-Assoc-Inst-
dc.titleIn vitro formation of active carboxypeptidase Y from pro-carboxypeptidase Y inclusion bodies by fed-batch operation-
dc.title.alternativeIn vitro formation of active carboxypeptidase Y from pro-carboxypeptidase Y inclusion bodies by fed-batch operation-
dc.typeArticle-
dc.citation.titleJournal of Microbiology and Biotechnology-
dc.citation.number5-
dc.citation.endPage889-
dc.citation.startPage887-
dc.citation.volume11-
dc.contributor.affiliatedAuthorMoon Sun Hahm-
dc.contributor.affiliatedAuthorBong Hyun Chung-
dc.contributor.alternativeName함문선-
dc.contributor.alternativeName정봉현-
dc.identifier.bibliographicCitationJournal of Microbiology and Biotechnology, vol. 11, no. 5, pp. 887-889-
dc.subject.keywordE. coli-
dc.subject.keywordpro-carboxypeptidase Y (pro-CPY)-
dc.subject.keywordcarboxypeptidase Y (CPY)-
dc.subject.keywordrefolding-
dc.subject.keywordproteinase K-
dc.subject.localEscherichia coli.-
dc.subject.localescherichia coli-
dc.subject.localEscherichia Coli-
dc.subject.localEscherichia coli-
dc.subject.localE.coli-
dc.subject.localescherichia coil-
dc.subject.localE. coli-
dc.subject.localE. Coli-
dc.subject.localpro-carboxypeptidase Y-
dc.subject.localpro-carboxypeptidase Y (pro-CPY)-
dc.subject.localcarboxypeptidase Y (CPY)-
dc.subject.localRefolding-
dc.subject.localrefolding-
dc.subject.localproteinase K-
dc.description.journalClassY-
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