title: Uncovering the formation and selection of benzylmalonyl-CoA from the biosynthesis of splenocin and enterocin reveals a versatile way to introduce amino acids into polyketide carbon scaffolds.
abstract: Selective modification of carbon scaffolds via biosynthetic engineering is important for polyketide structural diversification. Yet, this scope is currently restricted to simple aliphatic groups due to (1) limited variety of CoA-linked extender units, which lack aromatic structures and chemical reactivity, and (2) narrow acyltransferase (AT) specificity, which is limited to aliphatic CoA-linked extender units. In this report, we uncovered and characterized the first aromatic CoA-linked extender unit benzylmalonyl-CoA from the biosynthetic pathways of splenocin and enterocin in Streptomyces sp. CNQ431. Its synthesis employs a deamination/reductive carboxylation strategy to convert phenylalanine into benzylmalonyl-CoA, providing a link between amino acid and CoA-linked extender unit synthesis. By characterization of its selection, we further validated that AT domains of splenocin, and antimycin polyketide synthases are able to select this extender unit to introduce the phenyl group into their dilactone scaffolds. The biosynthetic machinery involved in the formation of this extender unit is highly versatile and can be potentially tailored for tyrosine, histidine and aspartic acid. The disclosed aromatic extender unit, amino acid-oriented synthetic pathway, and aromatic-selective AT domains provides a systematic breakthrough toward current knowledge of polyketide extender unit formation and selection, and also opens a route for further engineering of polyketide carbon scaffolds using amino acids.
authors.affiliation: †Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuhan 430071, China.
authors.forename: Chenchen
authors.initials: C
authors.lastname: Chang
authors.affiliation: †Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuhan 430071, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Huang
authors.affiliation: ‡State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai 200032, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yan
authors.affiliation: †Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuhan 430071, China.
authors.forename: Hongmin
authors.initials: H
authors.lastname: Ma
authors.affiliation: †Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuhan 430071, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Dai
authors.affiliation: †Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuhan 430071, China.
authors.forename: Benying
authors.initials: B
authors.lastname: Zhang
authors.affiliation: †Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuhan 430071, China.
authors.forename: Zixin
authors.initials: Z
authors.lastname: Deng
authors.affiliation: ‡State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai 200032, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Liu
authors.affiliation: †Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuhan 430071, China.
authors.forename: Xudong
authors.initials: X
authors.lastname: Qu
mesh_terms: D000217:Acyltransferases / Q000378:metabolism; D000968:Antimycin A / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D001593:Benzyl Compounds / Q000737:chemistry / Q000378:metabolism*; D053898:Biosynthetic Pathways; D001952:Bridged-Ring Compounds / Q000737:chemistry / Q000378:metabolism; D008316:Malonyl Coenzyme A / Q000737:chemistry / Q000378:metabolism*; D060847:Metabolic Engineering; D048630:Polyketide Synthases / Q000378:metabolism; D061065:Polyketides / Q000737:chemistry / Q000378:metabolism*; D013302:Streptomyces / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D001593:Benzyl Compounds; D001952:Bridged-Ring Compounds; D061065:Polyketides; D008316:Malonyl Coenzyme A; C012306:enterocin; D000968:Antimycin A; D048630:Polyketide Synthases; D000217:Acyltransferases