Acyltransferase mediated polyketide release from a fungal megasynthase.
Acyltransferase mediated polyketide release from a fungal megasynthase.
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DOI：
10.1021/ja903203g
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发表时间：
2009-06-24
影响因子：
15
通讯作者：
Tang, Yi
中科院分区：
文献类型：
--
作者：
Xie, Xinkai;Meehan, Michael J.;Xu, Wei;Dorrestein, Pieter C.;Tang, Yi
LOVF是丝状真菌土曲霉的高度还原聚酮合成酶(HR-PKS)。洛伐他汀合成α-S-甲基丁酸甲酯侧链，然后转移到莫纳可林J，产生降胆固醇的天然产品洛伐他汀。在该报告中，我们表达了全长的LovF，并在体外重组了巨合酶的活性。我们证实了LovF的二酮产物是通过解离的酰基转移酶LovD从LovF ACP结构域上卸载出来的。这是第一个由酰基转移酶介导的从真菌PKS释放聚酮产物的例子。我们确定LovD主要与LovF的ACP结构域相互作用，蛋白质-蛋白质相互作用导致二酮产物的高效转移。用LovF代替N-乙酰半胱胺作为酰基载体，催化效率提高了近百万倍。LovF卸载机制的重建和鉴定为了解真菌HR-PKS的功能提供了新的见解。
LovF is a highly reducing polyketide synthase (HR-PKS) from the filamentous fungus Aspergillus terreus. LovF synthesizes the α-S-methylbutyrate side chain that is subsequently transferred to monacolin J to yield the cholesterol-lowering natural product lovastatin. In the report, we expressed the full length LovF and reconstituted the megasynthase activities in vitro. We confirmed the diketide product of LovF is offloaded from the LovF ACP domain by the dissociated acyltransferase LovD. This represents the first example of acyltransferase-mediated release of polyketide products from fungal PKSs. We determined LovD primarily interacts with the ACP domain of LovF and the protein-protein interactions leads to highly efficient transfer of the diketide product. The catalytic efficiency is enhanced nearly one million-fold when LovF was used as the acyl carrier instead of N-acetylcysteamine. Reconstitution and characterization of the LovF offloading mechanism provide new insights into the functions of fungal HR-PKS.
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影响因子：
56.9
作者：
Maier, Timm;Leibundgut, Marc;Ban, Nenad
通讯作者：
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影响因子：
3.3
作者：
ENDO, A;HASUMI, K;MASUDA, M
通讯作者：
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影响因子：
2.9
作者：
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影响因子：
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作者：
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通讯作者：
Hutchinson, CR
DOI：
10.1073/pnas.011399198
发表时间：
2001-12-18
影响因子：
11.1
作者：
Tsai, SC;Miercke, LJW;Stroud, RM
通讯作者：
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