Gene cloning, sequence analysis, and expression of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase

Pimchai Chaiyen; David P. Ballou; Vincent Massey

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Gene cloning, sequence analysis, and expression of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase

机译：2-甲基-3-羟基吡啶-5-羧酸加氧酶的基因克隆，序列分析和表达

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摘要

The gene encoding 2-methyl-3-hydroxypyri- dine-5-carboxylic acid oxygenase (MHPCO; EC 1.14.12.4) was cloned by using an oligonucleotide probe corresponding to the N terminus of the enzyme to screen a DNA library of Pseudo- monas sp. MA-1. The gene encodes for a protein of 379 amino acid residues corresponding to a molecular mass of 41.7 kDa, the same as that previously estimated for MHPCO. MHPCO was expressed in Escherichia coli and found to have the same properties as the native enzyme from Pseudomonas sp. MA-1. This study shows that MHPCO is a homotetrameric protein with one flavin adenine dinucleotide bound per subunit. Sequence comparison of the enzyme with other hydroxylases reveals regions that are conserved among aromatic flavopro- tein hydroxylases.

机译：通过使用对应于该酶N末端的寡核苷酸探针克隆编码2-甲基-3-羟基吡啶-5-羧酸加氧酶的基因（MHPCO； EC 1.14.12.4），以筛选假单胞菌的DNA文库。 sp。 MA-1。该基因编码一个379个氨基酸残基的蛋白质，对应于41.7 kDa的分子量，与先前估计的MHPCO相同。 MHPCO在大肠杆菌中表达，发现与假单胞菌属的天然酶具有相同的特性。 MA-1。这项研究表明，MHPCO是一种同四聚体蛋白，每个亚基结合一个黄素腺嘌呤二核苷酸。该酶与其他羟化酶的序列比较揭示了芳香黄素羟化酶中保守的区域。

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《Proceedings of the National Academy of Sciences of the United States of America》 |1997年第14期|p.7233-7238|共6页
作者
Pimchai Chaiyen; David P. Ballou; Vincent Massey;
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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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正文语种 eng
中图分类自然科学总论;
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6. Gene cloning sequence analysis and expression of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase [O] . Pimchai Chaiyen, David P. Ballou, Vincent Massey 1997

机译：2-甲基-3-羟基吡啶-5-羧酸加氧酶的基因克隆序列分析和表达
7. Crystallization and preliminary X-ray crystallographic analysis of 2-methyl-3-hydroxypyridine-5-carboxylic acid (MHPC) oxygenase from Pseudomonas sp. MA-1 [O] . Oonanant, Worrapoj, Sucharitakul, Jeerus, Yuvaniyama, Jirundon, 2005

机译：假单胞菌2-甲基-3-羟基吡啶-5-羧酸（MHPC）加氧酶的结晶和初步X射线晶体学分析。 MA-1

Gene cloning, sequence analysis, and expression of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase

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