Biosynthesis of 4-methylproline in cyanobacteria:: Cloning of nosE and nosF genes and biochemical characterization of the encoded dehydrogenase and reductase activities
Biosynthesis of 4-methylproline in cyanobacteria:: Cloning of nosE and nosF genes and biochemical characterization of the encoded dehydrogenase and reductase activities
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DOI：
10.1021/jo026479q
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发表时间：
2003-01-10
影响因子：
3.6
通讯作者：
Moore, RE
中科院分区：
文献类型：
--
作者：
Luesch, H;Hoffmann, D;Moore, RE
从遗传水平和酶促水平研究了蓝藻Nostoc属罕见氨基酸4-甲基脯氨酸的生物合成。克隆了两个参与生物合成的基因，并在大肠杆菌中过表达了一个锌依赖的长链脱氢酶和一个δ(1)-吡咯-5-羧酸（P5C）还原酶同源物，并进行了生化表征。合成了假定底物来测试酶底物的特异性，并进行了氘标记研究，以揭示酶反应与底物以及辅酶的立体特异性。
The biosynthesis of the unusual amino acid 4-methylproline in the Nostoc genus of cyanobacteria was investigated on the genetic and enzymatic level. Two genes involved in the biosynthesis were cloned and the corresponding enzymes, a zinc-dependent long-chain dehydrogenase and a Delta(1)-pyrroline-5-carboxylic acid (P5C) reductase homologue, were overexpressed in Escherichia coli and biochemically characterized. Putative substrates were synthesized to test enzyme substrate specificities, and deuterium labeling studies were carried out to reveal the stereospecificities of the enzymatic reactions with respect to the substrates as well as to the coenzymes.