Insights into Heterocycle Biosynthesis in the Cytotoxic Polyketide Alkaloid Janustatin A from a Plant-Associated Bacterium.

IF 2.9 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemistry Biochemistry Pub Date : 2025-01-21 Epub Date: 2025-01-09 DOI:10.1021/acs.biochem.4c00542
Stefan Leopold-Messer, Pornsuda Chawengrum, Jörn Piel
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Abstract

Janustatin A is a potently cytotoxic polyketide alkaloid produced at trace amounts by the marine bacterial plant symbiont Gynuella sunshinyii. Its biosynthetic terminus features an unusual pyridine-containing bicyclic system of unclear origin, in which polyketide and amino acid extension units appear reversed compared to the order of enzymatic modules in the polyketide synthase (PKS)-nonribosomal peptide synthetase (NRPS) assembly line. To elucidate unknown steps in heterocycle formation, we first established robust genome engineering tools in G. sunshinyii. A combination of gene deletion, complementation, production improvement, and NMR experiments then demonstrated that two desaturase homologues, JanA and JanB, are involved in hydroxylation and pyridine formation by desaturation, respectively. Structure-activity relationship studies showed that these modifications substantially increase the cytotoxicity and that the fully functionalized heterocyclic system is crucial for sub-nanomolar cytotoxicity. Isolation of the early post-PKS intermediate janustatin D with an already reversed heterocycle topology supports a noncanonical rearrangement process occurring on the PKS-NRPS assembly line.

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植物相关细菌细胞毒性聚酮生物碱Janustatin A的杂环生物合成研究。
Janustatin A是一种极具细胞毒性的聚酮生物碱,由海洋细菌植物共生Gynuella sunshinyii产生。它的生物合成端具有不寻常的含吡啶双环系统,其来源不明,其中聚酮和氨基酸延伸单元与聚酮合成酶(PKS)-非核糖体肽合成酶(NRPS)装配线中的酶模块顺序相反。为了阐明杂环形成的未知步骤,我们首先在G. sunshinyii中建立了强大的基因组工程工具。基因缺失、互补、生产改进和核磁共振实验的结合表明,两个去饱和酶同源物JanA和JanB分别参与了羟基化和吡啶的去饱和形成。结构-活性关系研究表明,这些修饰大大增加了细胞毒性,并且充分功能化的杂环系统对亚纳摩尔细胞毒性至关重要。分离出具有反向杂环拓扑结构的早期pks后中间janustatin D,支持PKS-NRPS装配线上发生的非规范重排过程。
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来源期刊
Biochemistry Biochemistry
Biochemistry Biochemistry 生物-生化与分子生物学
CiteScore
5.50
自引率
3.40%
发文量
336
审稿时长
1-2 weeks
期刊介绍: Biochemistry provides an international forum for publishing exceptional, rigorous, high-impact research across all of biological chemistry. This broad scope includes studies on the chemical, physical, mechanistic, and/or structural basis of biological or cell function, and encompasses the fields of chemical biology, synthetic biology, disease biology, cell biology, nucleic acid biology, neuroscience, structural biology, and biophysics. In addition to traditional Research Articles, Biochemistry also publishes Communications, Viewpoints, and Perspectives, as well as From the Bench articles that report new methods of particular interest to the biological chemistry community.
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