Rapid characterization and engineering of natural product biosynthetic pathways via DNA assembler.

We report a synthetic biology strategy for rapid genetic manipulation of natural product biosynthetic pathways. Based on DNA assembler, this method synthesizes the entire expression vector containing the target biosynthetic pathway and the genetic elements required for DNA maintenance and replication in various hosts in a single-step manner through yeast homologous recombination, offering unprecedented flexibility and versatility in pathway manipulations.

[1]  A. Bull,et al.  Biodiversity as a source of innovation in biotechnology. , 1992, Annual review of microbiology.

[2]  N. Kelleher,et al.  Heterologous production of fosfomycin and identification of the minimal biosynthetic gene cluster. , 2006, Chemistry & biology.

[3]  K. Rinehart,et al.  Spectinabilin, a new nitro-containing metabolite isolated from streptomyces spectabilis , 1976 .

[4]  G. Lancini,et al.  Biosynthesis of Secondary Metabolites , 1993 .

[5]  J Craig Venter,et al.  One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome , 2008, Proceedings of the National Academy of Sciences.

[6]  N. Kelleher,et al.  Cloning, expression, and biochemical characterization of Streptomyces rubellomurinus genes required for biosynthesis of antimalarial compound FR900098. , 2008, Chemistry & biology.

[7]  H. Gross.  Strategies to unravel the function of orphan biosynthesis pathways: recent examples and future prospects , 2007, Applied Microbiology and Biotechnology.

[8]  Adrian Keatinge-Clay,et al.  Crystal structure of the erythromycin polyketide synthase dehydratase. , 2008, Journal of molecular biology.

[9]  J. Staunton,et al.  Polyketide biosynthesis: a millennium review. , 2001, Natural product reports.

[10]  S. Brady,et al.  Fluostatins produced by the heterologous expression of a TAR reassembled environmental DNA derived type II PKS gene cluster. , 2010, Journal of the American Chemical Society.

[11]  Thomas H Segall-Shapiro,et al.  Creation of a Bacterial Cell Controlled by a Chemically Synthesized Genome , 2010, Science.

[12]  Gregory L Challis,et al.  Mining microbial genomes for new natural products and biosynthetic pathways. , 2008, Microbiology.

[13]  D. Newman,et al.  Natural products as sources of new drugs over the last 25 years. , 2007, Journal of natural products.

[14]  C. Miranda,et al.  Deciphering Pactamycin Biosynthesis and Engineered Production of New Pactamycin Analogues , 2009, Chembiochem : a European journal of chemical biology.

[15]  V. Noskov,et al.  A genetic system for direct selection of gene-positive clones during recombinational cloning in yeast. , 2002, Nucleic acids research.

[16]  Zengyi Shao,et al.  DNA assembler, an in vivo genetic method for rapid construction of biochemical pathways , 2008, Nucleic acids research.

[17]  S. Eykyn.  Microbiology , 1950, The Lancet.

[18]  Eriko Takano,et al.  A cryptic type I polyketide synthase (cpk) gene cluster in Streptomyces coelicolor A3(2) , 2007, Archives of Microbiology.

[19]  Arnold L. Demain,et al.  From natural products discovery to commercialization: a success story , 2006, Journal of Industrial Microbiology and Biotechnology.

[20]  N. Kelleher,et al.  Unusual transformations in the biosynthesis of the antibiotic phosphinothricin tripeptide. , 2007, Nature chemical biology.

[21]  Jared R Leadbetter,et al.  Cultivation of recalcitrant microbes: cells are alive, well and revealing their secrets in the 21st century laboratory. , 2003, Current opinion in microbiology.

[22]  Andrew G. Glen,et al.  APPL , 2001 .

[23]  G. Challis,et al.  2-Alkyl-4-hydroxymethylfuran-3-carboxylic acids, antibiotic production inducers discovered by Streptomyces coelicolor genome mining , 2008, Proceedings of the National Academy of Sciences.

[24]  V. Noskov,et al.  A general cloning system to selectively isolate any eukaryotic or prokaryotic genomic region in yeast , 2003, BMC Genomics.

[25]  J. Vederas,et al.  Drug Discovery and Natural Products: End of an Era or an Endless Frontier? , 2009, Science.

[26]  T. Nonaka,et al.  Hidden Function of Catalytic Domain in 6-Methylsalicylic Acid Synthase for Product Release* , 2010, The Journal of Biological Chemistry.

[27]  A. Barberis,et al.  Coupling homologous recombination with growth selection in yeast: a tool for construction of random DNA sequence libraries. , 2004, BioTechniques.

[28]  Huimin Zhao,et al.  Cloning and heterologous expression of the spectinabilin biosynthetic gene cluster from Streptomyces spectabilis. , 2010, Molecular bioSystems.

[29]  Rolf Müller,et al.  A Type I/Type III Polyketide Synthase Hybrid Biosynthetic Pathway for the Structurally Unique ansa Compound Kendomycin , 2008, Chembiochem : a European journal of chemical biology.

[30]  C. Raymond,et al.  General method for plasmid construction using homologous recombination. , 1999, BioTechniques.

[31]  G. Challis,et al.  Strategies for the Discovery of New Natural Products by Genome Mining , 2009, Chembiochem : a European journal of chemical biology.

[32]  D. Schlessinger,et al.  Recovery and potential utility of YACs as circular YACs/BACs. , 2000, Nucleic acids research.

[33]  W. Kelly,et al.  Analysis of the Indanomycin Biosynthetic Gene Cluster from Streptomyces antibioticus NRRL 8167 , 2009, Chembiochem : a European journal of chemical biology.

[34]  P. Dewick,et al.  Medicinal Natural Products: A Biosynthetic Approach , 1997 .

[35]  J. Pernodet,et al.  Regulation of the Biosynthesis of the Macrolide Antibiotic Spiramycin in Streptomyces ambofaciens , 2010, Journal of bacteriology.

[36]  C. Hertweck,et al.  Iteration as programmed event during polyketide assembly; molecular analysis of the aureothin biosynthesis gene cluster. , 2003, Chemistry & biology.

[37]  M. Tanticharoen,et al.  Potent In Vitro Antimalarial Activity of Metacycloprodigiosin Isolated from Streptomycesspectabilis BCC 4785 , 2002, Antimicrobial Agents and Chemotherapy.

[38]  Elke Dittmann,et al.  Non‐Colinear Polyketide Biosynthesis in the Aureothin and Neoaureothin Pathways: An Evolutionary Perspective , 2007, Chembiochem : a European journal of chemical biology.