Enzymatic synthesis of resorcylic acid lactones by cooperation of fungal iterative polyketide synthases involved in hypothemycin biosynthesis.
Enzymatic synthesis of resorcylic acid lactones by cooperation of fungal iterative polyketide synthases involved in hypothemycin biosynthesis.
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DOI：
10.1021/ja100060k
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发表时间：
2010-04-07
影响因子：
15
通讯作者：
Tang, Yi
中科院分区：
文献类型：
--
作者：
Zhou, Hui;Qiao, Kangjian;Gao, Zhizeng;Meehan, Michael J.;Li, Jesse W. -H.;Zhao, Xiling;Dorrestein, Pieter C.;Vederas, John C.;Tang, Yi
Hypothemycin 是一种来自真菌 Hypomyces subiculosus 的大环内酯蛋白激酶抑制剂。在生物合成过程中，其碳框架由两种迭代聚酮合酶 (PKS)、Hpm8（高度还原性）和 Hpm3（非还原性）组装而成。这些在酿酒酵母 BJ5464-NpgA 中异源表达，纯化至接近同质并在体外重构，以从丙二酰辅酶 A 和 NADPH 产生 (6'S, 10'S)-反式-7',8'-脱氢玉米赤霉烯醇 (1)。 1的结构通过X射线晶体学分析确定。在缺乏功能性 Hpm3 的情况下，还原性 PKS Hpm8 会产生并卸载截短的吡喃酮产物，而不是预期的六酮化合物。非还原性 Hpm3 能够接受正确官能化的六酮化合物的 N-乙酰半胱胺硫酯以形成 1，但它无法启动丙二酰辅酶 A 形成聚酮化合物。我们表明，Hpm3 的起始单元酰基转移酶 (SAT) 对于两种酶之间的串扰至关重要，并且 1 的生物合成速率由 Hpm8 形成六酮化合物的速率决定。
Hypothemycin is a macrolide protein kinase inhibitor from the fungus Hypomyces subiculosus. During biosynthesis, its carbon framework is assembled by two iterative polyketide synthases (PKSs), Hpm8 (highly reducing) and Hpm3 (non-reducing). These were heterologously expressed in Saccharomyces cerevisiae BJ5464-NpgA, purified to near homogeneity and reconstituted in vitro to produce (6′S, 10′S)-trans-7′,8′-dehydrozearalenol (1) from malonyl-CoA and NADPH. The structure of 1 was determined by x-ray crystallographic analysis. In the absence of functional Hpm3, the reducing PKS Hpm8 produces and offloads truncated pyrone products instead of the expected hexaketide. The non-reducing Hpm3 is able to accept an N-acetylcysteamine thioester of a correctly functionalized hexaketide to form 1, but it is unable to initiate polyketide formation from malonyl-CoA. We show that the starter unit acyltransferase (SAT) of Hpm3 is critical for crosstalk between the two enzymes and that the rate of biosynthesis of 1 is determined by the rate of hexaketide formation by Hpm8.
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影响因子：
3
作者：
Rastelli, Giulio;Rosenfeld, Robin;Santi, Daniel V.
通讯作者：
Santi, Daniel V.
影响因子：
--
作者：
Watanabe, CMH;Townsend, CA
通讯作者：
Townsend, CA
影响因子：
4.4
作者：
Reeves, Christopher D.;Hu, Zhihao;Kealey, James T.
通讯作者：
Kealey, James T.
影响因子：
3.2
作者：
Crawford, Jason M.;Vagstad, Anna L.;Townsend, Craig A.
通讯作者：
Townsend, Craig A.
DOI：
10.1073/pnas.0800657105
发表时间：
2008-04-29
影响因子：
11.1
作者：
Zhou, Hui;Zhan, Jixun;Tang, Yi
通讯作者：
Tang, Yi