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Heterologous protein production using Psychrobacter sp. PAMC 21119 analyzed with a green fluorescent protein-based reporter system

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dc.contributor.author이민주-
dc.contributor.authorKim, Bomi-
dc.contributor.authorKim, Kitae-
dc.contributor.authorLee, Jun Hyuck-
dc.contributor.authorDo, Hackwon-
dc.date.accessioned2023-12-06T16:38:39Z-
dc.date.available2023-12-06T16:38:39Z-
dc.date.issued2023-
dc.identifier.urihttps://repository.kopri.re.kr/handle/201206/14918-
dc.description.abstractInsolubility and low expression are typical bottlenecks in the production of proteins for studying their function and structure using X-ray crystallography or nuclear magnetic resonance spectroscopy. Cold-active enzymes from polar microorganisms have unique structural features that render them unstable and thermolabile, and are responsible for decreased protein yield in heterologous expression systems. To address these challenges, we developed a heterologous protein expression system using a psychrophilic organism, Psychrobacter sp. PAMC 21119, as a protein expression host with its own promoter. We screened 11 promoters and evaluated their strength using quantitative real-time polymerase chain reaction and a reporter system harboring the SfGFP gene. The highest expression was achieved using promoters RH96_RS13655 (P21119_20930) and RH96_RS15090 (P21119_23410), regardless of the temperature used. The p20930 strain exhibited a maximum expression level 19.6-fold higher than that of its control at 20 °C and produced approximately 0.5 mg of protein per gram of dry cell weight. To our knowledge, this is the first report of a low-temperature recombinant protein expression system developed using Psychrobacter sp. that can be used to express various difficult-to-express and cold-active proteins.-
dc.languageEnglish-
dc.subject.classificationKing Sejong Station-
dc.titleHeterologous protein production using Psychrobacter sp. PAMC 21119 analyzed with a green fluorescent protein-based reporter system-
dc.title.alternative녹색 형광 단백질 기반의 리포터 시스템과 Psychrobacter sp. PAMC 21119을 이용한 이종 단백질 생산 분석-
dc.typeArticle-
dc.identifier.bibliographicCitation이민주, et al. 2023. "Heterologous protein production using Psychrobacter sp. PAMC 21119 analyzed with a green fluorescent protein-based reporter system". <em>PROTEIN EXPRESSION AND PURIFICATION</em>, 212(106352): 1-9.-
dc.citation.titlePROTEIN EXPRESSION AND PURIFICATION-
dc.citation.volume212-
dc.citation.number106352-
dc.identifier.doi10.1016/j.pep.2023.106352-
dc.citation.startPage1-
dc.citation.endPage9-
dc.description.articleClassificationSCIE-
dc.description.jcrRateJCR 2021:83.544-
dc.subject.keywordCold-active enzyme-
dc.subject.keywordPsychrobacter sp.-
dc.subject.keywordRecombinant protein expression-
dc.identifier.localId2023-0159-
Appears in Collections  
2023-2023, Development of potential antibiotic compounds using polar organism resources (23-23) / Lee, Jun Hyuck (PM23030)
2023-2023, Investigation of ice microstructure properties for developing low-temperature purification and environment/energy materials (23-23) / Kim, Kitae (PE23120)
2023-2023, Post-Polar Genomics Project: Functional genomic study for securing of polar useful genes (23-23) / Kim, Jin-Hyoung (PE23160)
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