Expression and purification of soluble and active human enterokinase light chain in Escherichia coli

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dc.contributor.authorYoung Su Kim-
dc.contributor.authorHye Jeong Lee-
dc.contributor.authorSang-Hyun Park-
dc.contributor.authorY C Kim-
dc.contributor.authorJungoh Ahn-
dc.date.accessioned2021-05-26T03:30:20Z-
dc.date.available2021-05-26T03:30:20Z-
dc.date.issued2021-
dc.identifier.issn2215017X-
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/24341-
dc.description.abstractHuman enterokinase light chain (hEKL) specifically cleaves the sequence (Asp)4-Lys↓X (D4K), making this a frequently used enzyme for site-specific cleavage of recombinant fusion proteins. However, hEKL production from Escherichia coli is limited due to intramolecular disulphide bonds. Here, we present strategies to obtain soluble and active hEKL from E. coli by expressing the hEKL variant C112S fused with maltose-binding protein (MBP) through D4K and molecular chaperons including GroEL/ES. The fusion protein self-cleaved in vivo, thereby removing the MBP in the E. coli cells. Thus, the self-cleaved hEKL variant was released into the culture medium. One-step purification using HisTrap™ chromatography purified the hEKL variant exhibiting an enzymatic activity of 3.1 × 103 U/mL (9.934 × 105 U/mg). The approaches presented here greatly simplify the purification of hEKL from E. coli without requiring refolding processes.-
dc.titleExpression and purification of soluble and active human enterokinase light chain in Escherichia coli-
dc.title.alternativeExpression and purification of soluble and active human enterokinase light chain in Escherichia coli-
dc.typeArticle-
dc.citation.titleBiotechnology Reports-
dc.citation.number0-
dc.citation.endPagee00626-
dc.citation.startPagee00626-
dc.citation.volume30-
dc.contributor.affiliatedAuthorYoung Su Kim-
dc.contributor.affiliatedAuthorHye Jeong Lee-
dc.contributor.affiliatedAuthorSang-Hyun Park-
dc.contributor.affiliatedAuthorJungoh Ahn-
dc.contributor.alternativeName김영수-
dc.contributor.alternativeName이혜정-
dc.contributor.alternativeName박상현-
dc.contributor.alternativeName김예천-
dc.contributor.alternativeName안정오-
dc.identifier.bibliographicCitationBiotechnology Reports, vol. 30, pp. e00626-e00626-
dc.identifier.doi10.1016/j.btre.2021.e00626-
dc.subject.keywordHuman enterokinase light chain-
dc.subject.keywordEscherichia coli-
dc.subject.keywordRecombinant protein-
dc.subject.keywordFusion technology-
dc.subject.keywordSelf-cleavage-
dc.subject.localEscherichia coli-
dc.subject.localRecombinant protein-
dc.description.journalClassN-
Appears in Collections:
Division of Bio Technology Innovation > BioProcess Engineering Center > 1. Journal Articles
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