Polyketide proofreading by an acyltransferase-like enzyme.

Polyketide proofreading by an acyltransferase-like enzyme.
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DOI:
10.1016/j.chembiol.2012.01.005
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发表时间:
2012-03-23
影响因子:
--
通讯作者:
Piel J
Piel J
中科院分区:
生物1区
文献类型:
--
作者:
Jensen K;Niederkrüger H;Zimmermann K;Vagstad AL;Moldenhauer J;Brendel N;Frank S;Pöplau P;Kohlhaas C;Townsend CA;Oldiges M;Hertweck C;Piel J

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反式酰基转移酶聚酮合成酶是一类重要的产生生物活性多酮的细菌酶。与教科书上的PKS的一个不同之处是存在一个或多个独立的AT样酶。虽然一个同源基因将丙二酰基单位加载到PKS上,但第二个拷贝(AT2)的功能尚不清楚。我们在一个未培养的共生菌中研究了参与Pederin生物合成的两个ATS PedC和PedD。PedD对不同的酰基载体蛋白(ACP)显示丙二酰基转移酶活性,但不显示乙酰转移酶活性。相反,AT2 PedC有效地水解了与N-乙酰半胱胺或ACP结合的酰基单元。它可以接受不同链长和官能化的底物,但不能裂解丙二酰ACP。这些数据与PedC在PKS校对中的作用一致,表明其他AT2同源物也具有类似的功能,并为聚酮滴度改进和生物合成研究提供了策略。
Trans-acyltransferase polyketide synthases (trans-AT PKSs) are an important group of bacterial enzymes producing bioactive polyketides. One difference from textbook PKSs is the presence of one or more free-standing AT-like enzymes. While one homolog loads the PKS with malonyl units, the function of the second copy (AT2) was unknown. We studied the two ATs PedC and PedD involved in pederin biosynthesis in an uncultivated symbiont. PedD displayed malonyl- but not acetyltransferase activity toward various acyl carrier proteins (ACPs). In contrast, the AT2 PedC efficiently hydrolyzed acyl units bound to N-acetylcysteamine or ACP. It accepted substrates with various chain lengths and functionalizations but did not cleave malonyl-ACP. These data are consistent with the role of PedC in PKS proofreading, suggesting a similar function for other AT2 homologs and providing strategies for polyketide titer improvement and biosynthetic investigations.
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