title: D-Alanylation in the Assembly of Ansatrienin Side Chain Is Catalyzed by a Modular NRPS.
abstract: Ansatrienins are a group of ansamycins with an N-cyclohexanoyl d-alanyl side chain. Though ansatrienins have been identified for decades, the mechanism for the addition of this unique side chain was not established. Here, we report the biochemical characterization of a tridomain nonribosomal peptide synthetase (NRPS), AstC, and an N-acyltransferase, AstF1, encoded in the biosynthetic pathway of ansatrienins. We demonstrate that AstC can efficiently catalyze the transfer of d-alanine to the C-11 hydroxyl group of ansatrienins, and AstF1 is able to attach the cyclohexanoyl group to the amino group of d-alanine. Remarkably, AstC presents the first example that a modular NRPS can catalyze intermolecular d-alanylation of the hydroxyl group to form an ester bond, though alanyl natural products have been known for decades. In addition, both AstC and AstF1 have broad substrate specificity toward acyl donors, which can be utilized to create novel ansatrienins.
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University , Jinan, Shandong 250100, Peoples' Republic of China.
authors.forename: Guoyin
authors.initials: G
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University , Jinan, Shandong 250100, Peoples' Republic of China.
authors.forename: Ning
authors.initials: N
authors.lastname: Shi
authors.affiliation: Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Sciences, Shandong University , Jinan, Shandong 250012, Peoples' Republic of China.
authors.forename: Yaoyao
authors.initials: Y
authors.lastname: Li
authors.affiliation: Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Sciences, Shandong University , Jinan, Shandong 250012, Peoples' Republic of China.
authors.forename: Wang
authors.initials: W
authors.lastname: Chen
authors.affiliation: Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Sciences, Shandong University , Jinan, Shandong 250012, Peoples' Republic of China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Deng
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University , Jinan, Shandong 250100, Peoples' Republic of China.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University , Jinan, Shandong 250100, Peoples' Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University , Jinan, Shandong 250100, Peoples' Republic of China.
authors.forename: Haoxin
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University , Jinan, Shandong 250100, Peoples' Republic of China.|Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Sciences, Shandong University , Jinan, Shandong 250012, Peoples' Republic of China.
authors.forename: Yuemao
authors.initials: Y
authors.lastname: Shen
mesh_terms: D000409:Alanine / Q000378:metabolism*; D002384:Catalysis; D047029:Lactams, Macrocyclic / Q000737:chemistry / Q000378:metabolism*
chemical_list: D047029:Lactams, Macrocyclic; D000409:Alanine