Inactivation of tyrosine phenol-lyase by Pictet-Spengler reaction and alleviation by T15A mutation on intertwined N-terminal arm = Pictet-Spengler 반응 Tyrosine Phenol-Lyase의 불활성화 및 N-말단 Arm

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dc.contributor.authorSeung Goo Lee-
dc.contributor.authorS P Hong-
dc.contributor.authorDo Young Kim-
dc.contributor.authorJae Jun Song-
dc.contributor.authorH S Ro-
dc.contributor.authorM H Sung-
dc.date.accessioned2017-04-19T09:05:36Z-
dc.date.available2017-04-19T09:05:36Z-
dc.date.issued2006-
dc.identifier.issn1742464X-
dc.identifier.uri10.1111/j.1742-4658.2006.05546.xko
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/7685-
dc.description.abstractCitrobacter freundii l-tyrosine phenol-lyase (TPL) was inactivated by a Pictet-Spengler reaction between the cofactor and a substrate, 3,4-dihydroxyphenyl-l-alanine (l-dopa), in proportion to an increase in the reaction temperature. Random mutagenesis of the tpl gene resulted in the generation of a Thr15 to Ala mutant (T15A), which exhibited a two-fold improved activity towards l-DOPA as the substrate. The Thr15 residue was located on the intertwined N-terminal arm of the TPL structure, and comprised an H-bond network in proximity to the hydrophobic core between the catalytic dimers. The maximum activity of the mutant and native enzymes with l-DOPA was detected at 45 and 40°C, respectively, which was 15°C lower than when using l-tyrosine as the substrate. The half-lives at 45°C were about 16.8 and 6.4 min for the mutant and native enzymes, respectively, in 10 mm l-DOPA. On treatment with excess pyridoxal-5′-phosphate (PLP), the l-DOPA-inactivated enzymes recovered over 80% of their original activities, thereby attributing the inactivation to a loss of the cofactor through Pictet-Spengler condensation with l-DOPA. Consistent with the extended half-life, the apparent Michaelis constant of the T15A enzyme for PLP (Km,PLP) increased slowly when increasing the temperature, while that of the native enzyme showed a sharp increase at temperatures higher than 50°C, implying that the loss of the cofactor with the Pictet-Spengler reaction was prevented by the tighter binding and smaller release of the cofactor in the mutant enzyme.-
dc.publisherWiley-
dc.titleInactivation of tyrosine phenol-lyase by Pictet-Spengler reaction and alleviation by T15A mutation on intertwined N-terminal arm = Pictet-Spengler 반응 Tyrosine Phenol-Lyase의 불활성화 및 N-말단 Arm-
dc.title.alternativeInactivation of tyrosine phenol-lyase by Pictet-Spengler reaction and alleviation by T15A mutation on intertwined N-terminal arm-
dc.typeArticle-
dc.citation.titleFEBS Journal-
dc.citation.number24-
dc.citation.endPage5573-
dc.citation.startPage5564-
dc.citation.volume273-
dc.contributor.affiliatedAuthorSeung Goo Lee-
dc.contributor.affiliatedAuthorDo Young Kim-
dc.contributor.affiliatedAuthorJae Jun Song-
dc.contributor.alternativeName이승구-
dc.contributor.alternativeName홍승표-
dc.contributor.alternativeName김도영-
dc.contributor.alternativeName송재준-
dc.contributor.alternativeName노현수-
dc.contributor.alternativeName성문희-
dc.identifier.bibliographicCitationFEBS Journal, vol. 273, no. 24, pp. 5564-5573-
dc.identifier.doi10.1111/j.1742-4658.2006.05546.x-
dc.subject.keywordCofactor affinity-
dc.subject.keywordL-DOPA-
dc.subject.keywordN-terminal arm-
dc.subject.keywordPictet-Spengler condensation-
dc.subject.keywordTyrosine phenol-lyase-
dc.subject.localCofactor affinity-
dc.subject.localL-DOPA-
dc.subject.localN-terminal arm-
dc.subject.localPictet-Spengler condensation-
dc.subject.localTyrosine phenol-lyase-
dc.description.journalClassY-
Appears in Collections:
Division of Biomaterials Research > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
Division of Biomaterials Research > Industrial Bio-materials Research Center > 1. Journal Articles
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
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