Purification, refolding, and characterization of recombinant Pseudomonas fluorescens lipase
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, KR | - |
dc.contributor.author | Kwon, DY | - |
dc.contributor.author | Yoon, SH | - |
dc.contributor.author | Kim, WY | - |
dc.contributor.author | Kim, KH | - |
dc.date.available | 2019-05-30T08:33:21Z | - |
dc.date.issued | 2005-01 | - |
dc.identifier.issn | 1046-5928 | - |
dc.identifier.issn | 1096-0279 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/24688 | - |
dc.description.abstract | Thermostable Pseudomonas fluorescens SIK W1 lipase (PFL), which is responsible for the spoilage of milk, was overexpressed as inclusion bodies in Escherichia coli. Renaturation of solubilized PFL was achieved by using size-exclusion protein refolding chromatography. The renatured enzyme was purified homogeneously using a combination of gel filtration and ion-exchange FPLC. Its specific activity was found to be enhanced in the presence of Ca2+. Secondary structural changes induced by Ca2+ were monitored by circular dichroism, which demonstrated that the activity increase of PFL in the presence of Ca2+ is strongly correlated with significant increases in alpha-helix and beta-sheet content. In the presence of Ca2+, the PFL structure was found resistant to denaturation by guanidine hydrochloride and to enzyme activity loss due to cosolvents like DMSO and trifluoroethanol, suggesting that Ca2+ plays an important role in inducing conformational changes and consequently in maintaining enzyme structural stability. (C) 2004 Elsevier Inc. All rights reserved. | - |
dc.format.extent | 6 | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | ACADEMIC PRESS INC ELSEVIER SCIENCE | - |
dc.title | Purification, refolding, and characterization of recombinant Pseudomonas fluorescens lipase | - |
dc.type | Article | - |
dc.identifier.doi | 10.1016/j.pep.2004.09.014 | - |
dc.identifier.bibliographicCitation | PROTEIN EXPRESSION AND PURIFICATION, v.39, no.1, pp 124 - 129 | - |
dc.description.isOpenAccess | N | - |
dc.identifier.wosid | 000226153600015 | - |
dc.identifier.scopusid | 2-s2.0-10644246117 | - |
dc.citation.endPage | 129 | - |
dc.citation.number | 1 | - |
dc.citation.startPage | 124 | - |
dc.citation.title | PROTEIN EXPRESSION AND PURIFICATION | - |
dc.citation.volume | 39 | - |
dc.type.docType | Article | - |
dc.publisher.location | 미국 | - |
dc.subject.keywordAuthor | refolding | - |
dc.subject.keywordAuthor | purification | - |
dc.subject.keywordAuthor | secondary structure | - |
dc.subject.keywordAuthor | lipase | - |
dc.subject.keywordAuthor | calcium ion | - |
dc.subject.keywordPlus | ESCHERICHIA-COLI | - |
dc.subject.keywordPlus | THERMOSTABLE LIPASE | - |
dc.subject.keywordPlus | BACTERIAL LIPASES | - |
dc.subject.keywordPlus | BACILLUS | - |
dc.subject.keywordPlus | GENE | - |
dc.subject.keywordPlus | CONFORMATION | - |
dc.subject.keywordPlus | AERUGINOSA | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | MILK | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Biotechnology & Applied Microbiology | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.relation.journalWebOfScienceCategory | Biotechnology & Applied Microbiology | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.
84, Heukseok-ro, Dongjak-gu, Seoul, Republic of Korea (06974)02-820-6194
COPYRIGHT 2019 Chung-Ang University All Rights Reserved.
Certain data included herein are derived from the © Web of Science of Clarivate Analytics. All rights reserved.
You may not copy or re-distribute this material in whole or in part without the prior written consent of Clarivate Analytics.