High throughput screening and directed evolution of tyrosine phenol-lyase = Tyrosine phenol-lyase의 고속탐색기술 개발 및 방향성 분자진화

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dc.contributor.authorSu-Lim Choi-
dc.contributor.authorEugene Rha-
dc.contributor.authorDo Young Kim-
dc.contributor.authorJae Jun Song-
dc.contributor.authorS P Hong-
dc.contributor.authorM H Sung-
dc.contributor.authorSeung Goo Lee-
dc.date.accessioned2017-04-19T09:04:18Z-
dc.date.available2017-04-19T09:04:18Z-
dc.date.issued2006-
dc.identifier.issn0257-2389-
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/7361-
dc.description.abstractRapid assay of enzyme is a primary requirement for successful application of directed evolution technology. Halo generation on a turbid plate would be a method of choice for high throughput screening of enzymes in this context. Here we report a new approach to prepare turbid plates, by controlling the crystallization of tyrosine to form needle-like particles. In the presence of tyrosine phenollyase (TPL), the needle-like tyrosine crystals were converted to soluble phenol rapidly than the usual rectangular tyrosine crystals. When an error-prone PCR library of Citrobacter freundii TPL was spread on the turbid plate, approximately 10% of the colonies displayed recognizable halos after 24 hours of incubation at 37°C. Representative positives from the turbid plates were transferred to LB-medium in 96-wellplates, cultivated overnight, and assayed for the enzyme activity with L-tyrosine as the substrate. The assay results were approximated to be proportional to the halo size on turbid plates, suggesting the screening system is directly applicable to the directed evolution of TPL. Actually, two best mutants on the turbid plates were identified to be 2-2.5 and 1.5-fold improved in the activity.-
dc.publisherKorea Soc-Assoc-Inst-
dc.titleHigh throughput screening and directed evolution of tyrosine phenol-lyase = Tyrosine phenol-lyase의 고속탐색기술 개발 및 방향성 분자진화-
dc.title.alternativeHigh throughput screening and directed evolution of tyrosine phenol-lyase-
dc.typeArticle-
dc.citation.titleKorean Journal of (Applied) Microbiology & Biotechnology-
dc.citation.number1-
dc.citation.endPage62-
dc.citation.startPage58-
dc.citation.volume34-
dc.contributor.affiliatedAuthorSu-Lim Choi-
dc.contributor.affiliatedAuthorEugene Rha-
dc.contributor.affiliatedAuthorDo Young Kim-
dc.contributor.affiliatedAuthorJae Jun Song-
dc.contributor.affiliatedAuthorSeung Goo Lee-
dc.contributor.alternativeName최수림-
dc.contributor.alternativeName나유진-
dc.contributor.alternativeName김도영-
dc.contributor.alternativeName송재준-
dc.contributor.alternativeName홍승표-
dc.contributor.alternativeName성문희-
dc.contributor.alternativeName이승구-
dc.identifier.bibliographicCitationKorean Journal of (Applied) Microbiology & Biotechnology, vol. 34, no. 1, pp. 58-62-
dc.subject.keyworderror-prone PCR-
dc.subject.keywordhigh throughput screening-
dc.subject.keywordneedle-like tyrosine crystal-
dc.subject.keywordturbid plate-
dc.subject.keywordtyrosine phenol-lyase-
dc.subject.keywordtyrosine phenol lyase-
dc.subject.keywordscreening-
dc.subject.localerror-prone PCR-
dc.subject.localHigh throughput screening-
dc.subject.localhigh throughput screening-
dc.subject.localHigh throughput screenig-
dc.subject.localneedle-like tyrosine crystal-
dc.subject.localturbid plate-
dc.subject.localTyrosine phenollyase-
dc.subject.localtyrosine phenol lyase-
dc.subject.localtyrosine phenol-lyase-
dc.subject.localTyrosine phenol-lyase-
dc.subject.localTyrosine phenollyase-
dc.subject.localtyrosine phenol lyase-
dc.subject.localtyrosine phenol-lyase-
dc.subject.localTyrosine phenol-lyase-
dc.subject.localScreening-
dc.subject.localscreening-
dc.description.journalClassN-
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
Division of Biomedical Research > Microbiome Convergence Research Center > 1. Journal Articles
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
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