Data_Sheet_1_New Alpiniamides From Streptomyces sp. IB2014/011-12 Assembled by an Unusual Hybrid Non-ribosomal Peptide Synthetase Trans-AT Polyketide Synthase Enzyme.docx

The environment of Lake Baikal is a well-known source of microbial diversity. The strain Streptomyces sp. IB2014/011-12, isolated from samples collected at Lake Baikal, was found to exhibit potent activity against Gram-positive bacteria. Here, we report isolation and characterization of linear polyketide alpiniamide A (1) and its new derivatives B–D (2–5). The structures of alpiniamides A–D were established and their relative configuration was determined by combination of partial Murata’s method and ROESY experiment. The absolute configuration of alpiniamide A was established through Mosher’s method. The gene cluster, responsible for the biosynthesis of alpiniamides (alp) has been identified by genome mining and gene deletion experiments. The successful expression of the cloned alp gene cluster in a heterologous host supports these findings. Analysis of the architecture of the alp gene cluster and the feeding of labeled precursors elucidated the alpiniamide biosynthetic pathway. The biosynthesis of alpiniamides is an example of a rather simple polyketide assembly line generating unusual chemical diversity through the combination of domain/module skipping and double bond migration events.

贝加尔湖的生境是公认的微生物多样性宝库。从贝加尔湖采集的样本中分离得到的链霉菌属（Streptomyces）菌株IB2014/011-12，被发现对革兰氏阳性菌具有强效抑菌活性。本文报道了线性聚酮类化合物alpiniamide A（1）及其新型衍生物B-D（2~5）的分离与表征。通过结合部分村田（Murata）分析法与ROESY（旋转坐标系核欧沃豪斯效应谱）实验，确定了alpiniamides A-D的结构及其相对构型。采用莫斯杰（Mosher）法确定了alpiniamide A的绝对构型。通过基因组挖掘与基因敲除实验，鉴定了负责alpiniamides生物合成的alp基因簇。将克隆得到的alp基因簇在异源宿主中成功表达，验证了上述研究结论。对alp基因簇的架构进行分析，并结合标记前体饲喂实验，阐明了alpiniamides的生物合成途径。alpiniamides的生物合成是一类较为简洁的聚酮组装线的典型实例，其通过结构域/模块跳跃与双键迁移事件的协同作用，产生了非同寻常的化学多样性。