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Targeted Discovery and Characterization of Type‐II PKS‐NRPS Hybrid DNA‐Alkylating Antibiotics

Journal content Created on 06 Aug 2025 by Angewandte Chemie Int Ed

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Angewandte Chemie Int Ed, Wiley-VCH

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Angewandte Chemie International Edition, EarlyView.

A new class of DNA‐alkylating antibiotics, xinghaicarcins, assembled by a type‐II PKS–NRPS hybrid system was discovered by self‐resistance gene‐directed genome mining. They possess an intricate spiro‐epoxide‐bearing spiroketal heptacyclic scaffold with a pipecolic acid, fully characterized by co‐crystallization. Xinghaicarcins show potent anticancer and antibacterial activity, which can be mitigated by resistance protein XhnU2. Abstract DNA alkylating natural products usually exhibit diverse bioactivity and serve as a crucial source of drug leads. Here, we employed genome mining guided by HTH‐42 superfamily resistance gene to precisely discover a new class of DNA‐alkylating antibiotics, xinghaicarcins, from Streptomyces xinghaiensis. They possess an intricate spiro‐epoxide‐bearing spiroketal heptacyclic scaffold fused with a pipecolic acid, assembled by a type II polyketide synthase–nonribosomal peptide synthetase hybrid system. An aminotransferase XhnB1 and a methyltransferase XhnM are identified to catalyze the formation of N‐methylated pipecolic acid building block, leading to the completion of the polyketide–peptide backbone. The identification of XhnM facilitated stereochemical determination of six chiral centers in xinghaicarcins by co‐crystallization. Notably, xinghaicarcins exhibit potent antibacterial activity against drug‐resistant pathogens and cytotoxicity against multiple cancer cell lines. Additionally, the HTH‐42 superfamily resistant protein, XhnU2, was characterized to mitigate xinghaicarcin‐induced genotoxicity. This work provides comprehensive insights into structure, biosynthesis, bioactivity, and self‐resistance mechanisms of xinghaicarcins, expanding diversity of DNA alkylating natural products. Targeted Discovery and Characterization of Type-II PKS-NRPS Hybrid DNA-Alkylating Antibiotics

A new class of DNA-alkylating antibiotics, xinghaicarcins, assembled by a type-II PKS–NRPS hybrid system was discovered by self-resistance gene-directed genome mining. They possess an intricate spiro-epoxide-bearing spiroketal heptacyclic scaffold with a pipecolic acid, fully characterized by co-crystallization. Xinghaicarcins show potent anticancer and antibacterial activity, which can be mitigated by resistance protein XhnU2.


Abstract

DNA alkylating natural products usually exhibit diverse bioactivity and serve as a crucial source of drug leads. Here, we employed genome mining guided by HTH-42 superfamily resistance gene to precisely discover a new class of DNA-alkylating antibiotics, xinghaicarcins, from Streptomyces xinghaiensis. They possess an intricate spiro-epoxide-bearing spiroketal heptacyclic scaffold fused with a pipecolic acid, assembled by a type II polyketide synthase–nonribosomal peptide synthetase hybrid system. An aminotransferase XhnB1 and a methyltransferase XhnM are identified to catalyze the formation of N-methylated pipecolic acid building block, leading to the completion of the polyketide–peptide backbone. The identification of XhnM facilitated stereochemical determination of six chiral centers in xinghaicarcins by co-crystallization. Notably, xinghaicarcins exhibit potent antibacterial activity against drug-resistant pathogens and cytotoxicity against multiple cancer cell lines. Additionally, the HTH-42 superfamily resistant protein, XhnU2, was characterized to mitigate xinghaicarcin-induced genotoxicity. This work provides comprehensive insights into structure, biosynthesis, bioactivity, and self-resistance mechanisms of xinghaicarcins, expanding diversity of DNA alkylating natural products.

Qiu‐Yue Nie, Shi‐Qi Fang, Lian Wu, Ruo‐Qin Gao, Yu Hu, Dian Ding, Hai‐Xue Pan, Zeng‐Fei Pei, Jun‐Bin He, Qiang Zhou, Zi‐Hui Chen, Xian‐Feng Hou, Xin‐Qing Zhao, Gong‐Li Tang

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