Plasmid Display for Stabilization of Enzymes Inside the Cell to Improve Whole-Cell Biotransformation Efficiency

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dc.contributor.authorPark, Yunjeongko
dc.contributor.authorShin, Jonghyeokko
dc.contributor.authorYang, Jinkyeongko
dc.contributor.authorKim, Hooyeonko
dc.contributor.authorJung, Younghunko
dc.contributor.authorOh, Hyunseokko
dc.contributor.authorKim, Yongjoonko
dc.contributor.authorHwang, Jaehyeonko
dc.contributor.authorPark, Myeongseoko
dc.contributor.authorBan, Choongjinko
dc.contributor.authorJeong, Ki Junko
dc.contributor.authorKim, Sun-Kiko
dc.contributor.authorKweon, Dae-Hyukko
dc.date.accessioned2020-02-11T06:20:31Z-
dc.date.available2020-02-11T06:20:31Z-
dc.date.created2020-02-10-
dc.date.created2020-02-10-
dc.date.created2020-02-10-
dc.date.created2020-02-10-
dc.date.created2020-02-10-
dc.date.issued2020-01-
dc.identifier.citationFRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v.7, pp.444-
dc.identifier.issn2296-4185-
dc.identifier.urihttp://hdl.handle.net/10203/272249-
dc.description.abstractRecombinant whole-cell biocatalysts are widely used for biotransformation of valuable products. However, some key enzymes involved in biotransformation processes are unstable and cannot be easily expressed in the functional form. In this study, we describe a versatile platform for enzyme stabilization inside the cell: Intracellularly Immobilized Enzyme System (IIES). A 1,2-fucosyltransferase from Pedobactor saltans (PsFL) and a 1,3-fucosyltransferase from Helicobacter pylori (HpFL), chosen as model proteins, were fused with Oct-1 DNA-binding domain, which mediated the formation of a plasmid-protein complex. Oct-1 fusion enabled both soluble and stable expression of recombinant proteins in the cytoplasm because the fusion proteins were stabilized on the plasmid like immobilized enzymes bound to solid surface. As a result, Oct-1-fusion proteins exhibited significantly greater product titer and yield than non-fusion proteins. Use of fusion proteins PsFL-Oct-1 with C-terminal Oct-1 and Oct-1-PsFL with N-terminal Oct-1 resulted in 3- and 2-fold higher 2 '-fucosyllactose titers, respectively, than with the use of PsFL alone. When Oct-1 was fused to HpFL, which requires dimerization through heptad repeats, almost two times more 3-fucosyllactose was produced. Fucosyllactose has been used as a food additive because it has various beneficial effects on human health. We anticipate that IIES using Oct-1 fusion protein developed in this study can be applied to stabilize other unstable enzymes.-
dc.languageEnglish-
dc.publisherFRONTIERS MEDIA SA-
dc.titlePlasmid Display for Stabilization of Enzymes Inside the Cell to Improve Whole-Cell Biotransformation Efficiency-
dc.typeArticle-
dc.identifier.wosid000509235900001-
dc.identifier.scopusid2-s2.0-85078399597-
dc.type.rimsART-
dc.citation.volume7-
dc.citation.beginningpage444-
dc.citation.publicationnameFRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY-
dc.identifier.doi10.3389/fbioe.2019.00444-
dc.contributor.localauthorJeong, Ki Jun-
dc.contributor.nonIdAuthorPark, Yunjeong-
dc.contributor.nonIdAuthorShin, Jonghyeok-
dc.contributor.nonIdAuthorYang, Jinkyeong-
dc.contributor.nonIdAuthorKim, Hooyeon-
dc.contributor.nonIdAuthorJung, Younghun-
dc.contributor.nonIdAuthorOh, Hyunseok-
dc.contributor.nonIdAuthorKim, Yongjoon-
dc.contributor.nonIdAuthorHwang, Jaehyeon-
dc.contributor.nonIdAuthorPark, Myeongseo-
dc.contributor.nonIdAuthorBan, Choongjin-
dc.contributor.nonIdAuthorKim, Sun-Ki-
dc.contributor.nonIdAuthorKweon, Dae-Hyuk-
dc.description.isOpenAccessY-
dc.type.journalArticleArticle-
dc.subject.keywordAuthorOct-1-
dc.subject.keywordAuthorDNA binding protein-
dc.subject.keywordAuthorintracellularly immobilized enzyme system-
dc.subject.keywordAuthorEscherichia coli-
dc.subject.keywordAuthorwhole-cell biotransformation-
dc.subject.keywordPlusDNA-BINDING DOMAIN-
dc.subject.keywordPlusESCHERICHIA-COLI-
dc.subject.keywordPlusHELICOBACTER-PYLORI-
dc.subject.keywordPlusENHANCED PRODUCTION-
dc.subject.keywordPlusPROTEIN EXPRESSION-
dc.subject.keywordPlusINCLUSION-BODIES-
dc.subject.keywordPlusPOU-DOMAIN-
dc.subject.keywordPlusTECHNOLOGY-
dc.subject.keywordPlusMETABOLISM-
dc.subject.keywordPlusMODULATION-
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