Effective Generation of Glucosylpiericidins with Selective Cytotoxicities and Insights into Their Biosynthesis
Effective Generation of Glucosylpiericidins with Selective Cytotoxicities and Insights into Their Biosynthesis
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DOI：
10.1128/aem.00294-21
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发表时间：
2021-07-01
影响因子：
4.4
通讯作者：
Li, Wenli
中科院分区：
文献类型：
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作者：
Liu, Zengzhi;Xiao, Fei;Li, Wenli
探索未知的糖基转移酶（GT）是重要的化合物结构的糖多样化在寻找候选药物。已报道杀粉蝶菌素糖苷具有有效的生物活性;然而，负责杀粉蝶菌素葡糖基化的GT仍然未知。在本文中，发现BmmGT 1（一种具有广泛底物选择性并从甲基营养型芽孢杆菌B-9987中分离的大环内酯GT）能够在体外对粉蝶菌素Al进行葡萄糖基化。接下来，基于BmmGT 1设计的密码子优化的GT基因sbmGT 1在杀粉蝶菌素生产者尤苏菲链霉菌（Streptomyces youssoufiensis）OUC 6819中异源表达。因此，杀粉蝶素糖苷显著积累，导致鉴定出四种新的glucopiericidin（化合物3、4、6和7）。进一步以BmmGT 1为探针，在S. youssoufiensis OUC 6819中发现的，并证明与杀粉蝶菌素葡糖基化相关;该基因的过表达导致鉴定出另一种新的杀粉蝶菌素糖苷，N-乙酰葡糖胺-杀粉蝶菌素（化合物8）。与THP-1淋巴瘤细胞系相比，化合物4、7和8显示出对A549、A375、HCT-116和111-29实体癌细胞系的细胞毒性选择性。此外，GT 1507同系物的数据库挖掘揭示了它们在细菌中的广泛分布，主要是在那些属于高GC革兰氏阳性菌和厚壁菌门的分支，从而代表了潜在的识别新的工具酶的化合物glycodifferentiations.IMPORTANCE许多生物活性的天然产物附加糖部分，往往是他们的生物活性的关键。糖基转移酶是天然产物糖基化的有力工具。虽然杀粉蝶菌素糖苷显示出有效的生物活性，但参与葡萄糖基化的GT尚不清楚。在这项研究中，五个新的杀粉蝶菌素糖苷（化合物3，4，6，7，和8）的GT编码基因的过表达后，产生的杀粉蝶菌素生产商。其中3个化合物（化合物4、7和8）显示出细胞毒性选择性。值得注意的是，GT 1507被证明与体内粉蝶菌素葡糖基化相关。此外，从GenBank数据库中挖掘GT 1507同源物揭示了它们在许多细菌中的广泛分布。我们的研究结果将极大地促进探索GT糖多样化的小分子在寻找候选药物。
Exploring unknown glycosyltransferases (GTs) is important for compound structural glycodiversification during the search for drug candidates. Piericidin glycosides have been reported to have potent bioactivities; however, the GT responsible for piericidin glucosylation remains unknown. Herein, BmmGT1, a macrolide GT with broad substrate selectivity and isolated from Bacillus methylotrophicus B-9987, was found to be able to glucosylate piericidin Al in vitro. Next, the codon-optimized GT gene sbmGTl, which was designed based on BmmGT1, was heterologously expressed in the piericidin producer Streptomyces youssoufiensis OUC6819. Piericidin glycosides thus significantly accumulated, leading to the identification of four new glucopiericidins (compounds 3, 4, 6, and 7). Furthermore, using BmmGT1 as the probe, GT1507 was identified in the genome of S. youssoufiensis OUC6819 and demonstrated to be associated with piericidin glucosylation; the overexpression of this gene led to the identification of another new piericidin glycoside, N-acetylglucosamine-piericidin (compound 8). Compounds 4, 7, and 8 displayed cytotoxic selectivity toward A549, A375, HCT-116, and 111-29 solid cancer cell lines compared to the THP-1 lymphoma cell line. Moreover, database mining of GT1507 homologs revealed their wide distribution in bacteria, mainly in those belonging to the high-GC Grampositive and Firmicutes clades, thus representing the potential for identification of novel tool enzymes for compound glycodiversification.IMPORTANCE Numerous bioactive natural products are appended with sugar moieties and are often critical for their bioactivities. Glycosyltransferases (GTs) are powerful tools for the glycodiversification of natural products. Although piericidin glycosides display potent bioactivities, the GT involved in glucosylation is unclear. In this study, five new piericidin glycosides (compounds 3, 4, 6, 7, and 8) were generated following the overexpression of GT-coding genes in a piericidin producer. Three of them (compounds 4, 7, and 8) displayed cytotoxic selectivity. Notably, GT1507 was demonstrated to be related to piericidin glucosylation in vivo. Furthermore, mining of GT1507 homologs from the GenBank database revealed their wide distribution across numerous bacteria. Our findings would greatly facilitate the exploration of GTs to glycodiversify small molecules in the search for drug candidates.