An FAD-Dependent Pyridine Nucleotide-Disulfide Oxidoreductase Is Involved in Disulfide Bond Formation in FK228 Anticancer Depsipeptide
An FAD-Dependent Pyridine Nucleotide-Disulfide Oxidoreductase Is Involved in Disulfide Bond Formation in FK228 Anticancer Depsipeptide
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DOI：
10.1016/j.chembiol.2009.05.005
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发表时间：
2009-06-26
影响因子：
--
通讯作者：
Cheng, Yi-Qiang
中科院分区：
文献类型：
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作者：
Wang, Cheng;Wesener, Shane R.;Cheng, Yi-Qiang
二硫键在细菌天然产物中是罕见的，在这些产物中形成二硫键的机制尚不清楚。在这里，我们描述了一个基因和它的产物关键的二硫键形成的FK228抗癌肽在紫色色杆菌。缺失depH显著降低了FK228的产量，而在中等拷贝数的质粒上与depH缺失突变体互补不仅完全恢复了FK228的产量，而且显著提高了FK228的产量。纯化的天然形式的6xhis标记的deep h融合蛋白是71.0 kDa的同二聚体，每个单体含有一个FAD分子。在NADP+存在的情况下，DepH有效地将FK228的直接前体转化为FK228。我们得出结论，DepH是一种fad依赖的吡啶核苷酸二硫氧化还原酶，特异性和有效地催化FK228中二硫键的形成。
Disulfide bonds are rare in bacterial natural products, and the mechanism of disulfide bond formation in those products is unknown. Here we characterize a gene and its product critical for a disulfide bond formation in FK228 anticancer depsipeptide in Chromobacterium violaceum. Deletion of depH drastically reduced FK228 production, whereas complementation of the depH-deletion mutant with a copy of depH on a medium copy-number plasmid not only fully restored the FK228 production but also significantly increased the FK228 yield. Purified 6xHis-tagged DepH fusion protein in native form is a homodimer of 71.0 kDa, with each monomer containing one molecule of FAD. DepH efficiently converts an immediate FK228 precursor to FK228 in the presence of NADP+. We conclude that DepH is an FAD-dependent pyridine nucleotide-disulfide oxidoreductase, specifically and efficiently catalyzing a disulfide bond formation in FK228.