Bioinformatic and Functional Evaluation of Actinobacterial Piperazate Metabolism
Bioinformatic and Functional Evaluation of Actinobacterial Piperazate Metabolism
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DOI：
10.1021/acschembio.8b01086
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发表时间：
2019-04-01
影响因子：
4
通讯作者：
Blodgett, Joshua A. V.
中科院分区：
文献类型：
--
作者：
Hu, Yifei;Qi, Yunci;Blodgett, Joshua A. V.
阿扎酯（Piz）是一种非蛋白质氨基酸，以其不寻常的N-N键基序而闻名。Piz是脯氨酸模拟物，其赋予肽构象刚性。因此，哌嗪基分子通常是生物活性的，并且是治疗探索所期望的。库茨奈氏菌属酶KtzI和KtzT的体外表征最近导致了Piz的生物合成途径。然而，体内的Piz analgesia仍然完全没有特征。在此，我们描述了使用生物信息学、遗传学和选择生物化学相结合对放线菌Piz代谢的系统询问。在黄链霉菌，变铅青链霉菌，和几个环境链霉菌菌株的研究，我们的数据表明，KtzI型酶有条件地抑制Piz生产。我们还首次证明了使用工程放线菌生产Piz单体的可行性。最后，我们表明，一些放线菌采用融合KtzI KtzT嵌合酶产生Piz。我们的研究结果对未来哌嗪类药物的发现、途径工程和精细化学品的生物生产具有重要意义。
Piperazate (Piz) is a nonproteinogenic amino acid noted for its unusual N N bond motif. Piz is a proline mimic that imparts conformational rigidity to peptides. Consequently, piperazyl molecules are often bioactive and desirable for therapeutic exploration. The in vitro characterization of Kutzneria enzymes KtzI and KtzT recently led to a biosynthetic pathway for Piz. However, Piz anabolism in vivo has remained completely uncharacterized. Herein, we describe the systematic interrogation of actinobacterial Piz metabolism using a combination of bioinformatics, genetics, and select biochemistry. Following studies in Streptomyces flaveolus, Streptomyces lividans, and several environmental Streptomyces isolates, our data suggest that KtzI-type enzymes are conditionally dispensable for Piz production. We also demonstrate the feasibility of Piz monomer production using engineered actinobacteria for the first time. Finally, we show that some actinobacteria employ fused KtzI KtzT chimeric enzymes to produce Piz. Our findings have implications for future piperazyl drug discovery, pathway engineering, and fine chemical bioproduction.