title: Diversity-oriented combinatorial biosynthesis of benzenediol lactone scaffolds by subunit shuffling of fungal polyketide synthases.
abstract: Combinatorial biosynthesis aspires to exploit the promiscuity of microbial anabolic pathways to engineer the synthesis of new chemical entities. Fungal benzenediol lactone (BDL) polyketides are important pharmacophores with wide-ranging bioactivities, including heat shock response and immune system modulatory effects. Their biosynthesis on a pair of sequentially acting iterative polyketide synthases (iPKSs) offers a test case for the modularization of secondary metabolic pathways into "build-couple-pair" combinatorial synthetic schemes. Expression of random pairs of iPKS subunits from four BDL model systems in a yeast heterologous host created a diverse library of BDL congeners, including a polyketide with an unnatural skeleton and heat shock response-inducing activity. Pairwise heterocombinations of the iPKS subunits also helped to illuminate the innate, idiosyncratic programming of these enzymes. Even in combinatorial contexts, these biosynthetic programs remained largely unchanged, so that the iPKSs built their cognate biosynthons, coupled these building blocks into chimeric polyketide intermediates, and catalyzed intramolecular pairing to release macrocycles or α-pyrones. However, some heterocombinations also provoked stuttering, i.e., the relaxation of iPKSs chain length control to assemble larger homologous products. The success of such a plug and play approach to biosynthesize novel chemical diversity bodes well for bioprospecting unnatural polyketides for drug discovery.
authors: Biotechnology Research Institute, The Chinese Academy of Agricultural Sciences, Beijing 100081, P. R. China; Natural Products Center, School of Natural Resources and the Environment, University of Arizona, Tucson, AZ 85706;
authors: Yuquan
authors: Y
authors: Xu
authors: Department of Biological Engineering, Utah State University, Logan, UT 84322; and.
authors: Tong
authors: T
authors: Zhou
authors: Department of Biological Engineering, Utah State University, Logan, UT 84322; and.
authors: Shuwei
authors: S
authors: Zhang
authors: Natural Products Center, School of Natural Resources and the Environment, University of Arizona, Tucson, AZ 85706;
authors: Patricia
authors: P
authors: Espinosa-Artiles
authors: Department of Biological Engineering, Utah State University, Logan, UT 84322; and.
authors: Luoyi
authors: L
authors: Wang
authors: Biotechnology Research Institute, The Chinese Academy of Agricultural Sciences, Beijing 100081, P. R. China;
authors: Wei
authors: W
authors: Zhang
authors: Biotechnology Research Institute, The Chinese Academy of Agricultural Sciences, Beijing 100081, P. R. China;
authors: Min
authors: M
authors: Lin
authors: Natural Products Center, School of Natural Resources and the Environment, University of Arizona, Tucson, AZ 85706; Bio5 Institute, University of Arizona, Tucson, AZ 85721.
authors: A A Leslie
authors: AA
authors: Gunatilaka
authors: Department of Biological Engineering, Utah State University, Logan, UT 84322; and jixun.zhan@usu.edu imolnar@email.arizona.edu.
authors: Jixun
authors: J
authors: Zhan
authors: Natural Products Center, School of Natural Resources and the Environment, University of Arizona, Tucson, AZ 85706; Bio5 Institute, University of Arizona, Tucson, AZ 85721 jixun.zhan@usu.edu imolnar@email.arizona.edu.
authors: István
authors: I
authors: Molnár
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D018869:Heat-Shock Response / Q000187:drug effects; D006801:Humans; D007783:Lactones / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D048630:Polyketide Synthases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D061065:Polyketides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D007783:Lactones; D061065:Polyketides; D021122:Protein Subunits; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D048630:Polyketide Synthases
keywords: fungal genetics; secondary metabolites