Biosynthesis of Phenylnannolone A, a Multidrug Resistance Reversal Agent from the Halotolerant Myxobacterium Nannocystis pusilla B150
Biosynthesis of Phenylnannolone A, a Multidrug Resistance Reversal Agent from the Halotolerant Myxobacterium Nannocystis pusilla B150
复制标题
DOI：
10.1002/cbic.201300676
复制
发表时间：
2014-03-21
期刊：
影响因子：
3.2
通讯作者：
Koenig, Gabriele M.
中科院分区：
文献类型：
--
作者：
Bouhired, Sarah M.;Cruesemann, Max;Koenig, Gabriele M.
粘细菌菌株微小微囊藻B150合成结构上新的聚酮化合物苯基南诺酮A-C。除了一些常见的挥发性物质和铁载体外，这些是来自微囊藻属的第一批天然产物。Phenylnannolone A显示对ABCB 1基因产物P-糖蛋白的抑制活性，并逆转癌细胞中的柔红霉素抗性。为了解释导致苯基甘露醇酮A形成的生化反应，鉴定了推定的生物合成基因（phn 1，phn 2）。Phn 2是一种具有NRPS样装载模块的聚酮合酶（PKS），其结构域顺序与苯基甘露醇酮A结构一致。通过-O-18（4）-ATP焦磷酸交换和磷酸泛酰巯基乙胺排出测定来确定装载模块的功能性和底物选择性。一个特定的激活肉桂酸的AMP连接酶进行检测。Phn 1是一种假定的丁酰辅酶A羧化酶（BCC），提供乙基丙二酰辅酶A，用于形成苯基甘露醇酮A的乙基取代部分。Phn 1是在乙基取代的天然化合物的生物合成基因中发现的第一个BCC。因此，由phn 1和phn 2编码的puristone的生物合成利用了第一个生物合成机制，其中涉及BCC和PKS。
The myxobacterial strain Nannocystis pusilla B150 synthesizes the structurally new polyketides phenylnannolone A-C. Apart from some common volatiles and siderophores, these are the first natural products from the genus Nannocystis. Phenylnannolone A shows inhibitory activity towards the ABCB1 gene product P-glycoprotein and reverses daunorubicin resistance in cancer cells. To decipher the biochemical reactions leading to the formation of phenylnannolone A, the putative biosynthetic genes were identified (phn1, phn2). Phn2 is a polyketide synthase (PKS) with an NRPS-like loading module, and its domain order is consistent with the phenylnannolone A structure. The functionality and substrate selectivity of the loading module were determined by means of a -O-18(4)-ATP pyrophosphate exchange and a phosphopantetheine ejection assay. A specific activation of cinnamic acid by the AMP-ligase was detected. Phn1 is a putative butyryl-CoA carboxylase (BCC), providing ethylmalonyl-CoA for the formation of the ethyl-substituted part of phenylnannolone A. Phn1 is the first BCC found in biosynthetic genes for an ethyl-substituted natural compound. Biosynthesis of phenylnannolone A, putatively encoded by phn1 and phn2, thus utilizes the first biosynthetic machinery in which both a BCC and a PKS are involved.