{"title":"皮毛霉苯丙氨酸解氨酶的纯化及特性研究。","authors":"Andrea Goldson-Barnaby, Christine H Scaman","doi":"10.1155/2013/670702","DOIUrl":null,"url":null,"abstract":"<p><p>Trichosporon cutaneum phenylalanine ammonia lyase was selected as a model to investigate the dual substrate activity of this family of enzymes. Sequencing of the PAL gene identified an extensive intron region at the N-terminus. Five amino acid residues differing from a prior report were identified. Highest Phe : Tyr activities (1.6 ± 0.3 : 0.4 ± 0.1 μ mol/h g wet weight) were induced by Tyr. The enzyme has a temperature optimum of 32°C and a pH optimum of 8-8.5 and shows no metal cofactor dependence. Michaelis-Menten kinetics (Phe, K m 5.0 ± 1.1 mM) and positive allostery (Tyr, K' 2.4 ± 0.6 mM, Hill coefficient 1.9 ± 0.5) were observed. Anion exchange chromatography gave a purification fold of 50 with 20% yield. The His-Gln motif (substrate selectivity switch region) indicates the enzyme's ability to act on both substrates. </p>","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":" ","pages":"670702"},"PeriodicalIF":0.0000,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2013/670702","citationCount":"17","resultStr":"{\"title\":\"Purification and Characterization of Phenylalanine Ammonia Lyase from Trichosporon cutaneum.\",\"authors\":\"Andrea Goldson-Barnaby, Christine H Scaman\",\"doi\":\"10.1155/2013/670702\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Trichosporon cutaneum phenylalanine ammonia lyase was selected as a model to investigate the dual substrate activity of this family of enzymes. Sequencing of the PAL gene identified an extensive intron region at the N-terminus. Five amino acid residues differing from a prior report were identified. Highest Phe : Tyr activities (1.6 ± 0.3 : 0.4 ± 0.1 μ mol/h g wet weight) were induced by Tyr. The enzyme has a temperature optimum of 32°C and a pH optimum of 8-8.5 and shows no metal cofactor dependence. Michaelis-Menten kinetics (Phe, K m 5.0 ± 1.1 mM) and positive allostery (Tyr, K' 2.4 ± 0.6 mM, Hill coefficient 1.9 ± 0.5) were observed. Anion exchange chromatography gave a purification fold of 50 with 20% yield. The His-Gln motif (substrate selectivity switch region) indicates the enzyme's ability to act on both substrates. </p>\",\"PeriodicalId\":11835,\"journal\":{\"name\":\"Enzyme Research\",\"volume\":\" \",\"pages\":\"670702\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2013-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1155/2013/670702\",\"citationCount\":\"17\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Enzyme Research\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1155/2013/670702\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2013/9/12 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Enzyme Research","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1155/2013/670702","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2013/9/12 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
Purification and Characterization of Phenylalanine Ammonia Lyase from Trichosporon cutaneum.
Trichosporon cutaneum phenylalanine ammonia lyase was selected as a model to investigate the dual substrate activity of this family of enzymes. Sequencing of the PAL gene identified an extensive intron region at the N-terminus. Five amino acid residues differing from a prior report were identified. Highest Phe : Tyr activities (1.6 ± 0.3 : 0.4 ± 0.1 μ mol/h g wet weight) were induced by Tyr. The enzyme has a temperature optimum of 32°C and a pH optimum of 8-8.5 and shows no metal cofactor dependence. Michaelis-Menten kinetics (Phe, K m 5.0 ± 1.1 mM) and positive allostery (Tyr, K' 2.4 ± 0.6 mM, Hill coefficient 1.9 ± 0.5) were observed. Anion exchange chromatography gave a purification fold of 50 with 20% yield. The His-Gln motif (substrate selectivity switch region) indicates the enzyme's ability to act on both substrates.
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