Effects of deletions of mbtH-like genes on clorobiocin biosynthesis in Streptomyces coelicolor.

In the biosynthetic gene cluster of the aminocoumarin antibiotic clorobiocin, the small ORF cloY encodes a 71 aa protein which shows significant sequence similarity to mbtH from the mycobactin biosynthetic gene cluster of Mycobacterium tuberculosis. mbtH-like genes are frequently found in the biosynthetic gene clusters of peptide antibiotics and siderophores, but their function has remained enigmatic. In a recent publication it has been suggested that these genes may have no function for secondary metabolite biosynthesis. An in-frame deletion of cloY in the clorobiocin cluster has now been carried out. When the modified cluster was expressed in the heterologous host Streptomyces coelicolor M512, clorobiocin was still formed. However, when the two further mbtH-like genes from elsewhere in the host genome were inactivated as well, clorobiocin formation was reduced dramatically. Complementation with cloY or with any of three other mbtH-like genes restored clorobiocin formation. This is the first report proving the requirement of an mbtH-like gene for secondary metabolite formation, and the first proof that different mbtH-like genes can functionally replace each other. Feeding of an mbtH-defective triple mutant strain with an intact 3-amino-4,7-dihydroxy-coumarin moiety restored antibiotic production, showing that cloY is specifically required for the formation of this moiety of the clorobiocin molecule.

[1]  C. Rausch,et al.  The small MbtH-like protein encoded by an internal gene of the balhimycin biosynthetic gene cluster is not required for glycopeptide production. , 2006, FEMS microbiology letters.

[2]  N. Magarvey,et al.  Biosynthetic Pathway for Mannopeptimycins, Lipoglycopeptide Antibiotics Active against Drug-Resistant Gram-Positive Pathogens , 2006, Antimicrobial Agents and Chemotherapy.

[3]  Mark S. Thomas,et al.  The Ornibactin Biosynthesis and Transport Genes of Burkholderia cenocepacia Are Regulated by an Extracytoplasmic Function σ Factor Which Is a Part of the Fur Regulon , 2006, Journal of bacteriology.

[4]  Ruth H Flatman,et al.  Structure-Activity Relationships of Aminocoumarin-Type Gyrase and Topoisomerase IV Inhibitors Obtained by Combinatorial Biosynthesis , 2006, Antimicrobial Agents and Chemotherapy.

[5]  F. de la Calle,et al.  Deciphering the Biosynthesis Pathway of the Antitumor Thiocoraline from a Marine Actinomycete and Its Expression in Two Streptomyces Species , 2006, Chembiochem : a European journal of chemical biology.

[6]  Vivian Miao,et al.  The lipopeptide antibiotic A54145 biosynthetic gene cluster from Streptomyces fradiae , 2006, Journal of Industrial Microbiology and Biotechnology.

[7]  L. Heide,et al.  Acyl Transfer in Clorobiocin Biosynthesis: Involvement of Several Proteins in the Transfer of the Pyrrole‐2‐carboxyl Moiety to the Deoxysugar , 2005, Chembiochem : a European journal of chemical biology.

[8]  C. Walsh,et al.  NovJ/NovK catalyze benzylic oxidation of a beta-hydroxyl tyrosyl-S-pantetheinyl enzyme during aminocoumarin ring formation in novobiocin biosynthesis. , 2005, Biochemistry.

[9]  L. Heide,et al.  NovG, a DNA-binding protein acting as a positive regulator of novobiocin biosynthesis. , 2005, Microbiology.

[10]  K. Chater,et al.  Heterologous Expression of Novobiocin and Clorobiocin Biosynthetic Gene Clusters , 2005, Applied and Environmental Microbiology.

[11]  Christopher J. Silva,et al.  Daptomycin biosynthesis in Streptomyces roseosporus: cloning and analysis of the gene cluster and revision of peptide stereochemistry. , 2005, Microbiology.

[12]  P. Dorrestein,et al.  Characterization of the formation of the pyrrole moiety during clorobiocin and coumermycin A1 biosynthesis. , 2005, Biochemistry.

[13]  A. Maxwell,et al.  Simocyclinone D8, an Inhibitor of DNA Gyrase with a Novel Mode of Action , 2005, Antimicrobial Agents and Chemotherapy.

[14]  L. Heide,et al.  New aminocoumarin antibiotics from genetically engineered Streptomyces strains. , 2005, Current medicinal chemistry.

[15]  W. Metcalf,et al.  Molecular Cloning, Sequence Analysis, and Heterologous Expression of the Phosphinothricin Tripeptide Biosynthetic Gene Cluster from Streptomyces viridochromogenes DSM 40736 , 2005, Antimicrobial Agents and Chemotherapy.

[16]  S. Berger,et al.  Biosynthetic Gene Cluster of the Herbicide Phosphinothricin Tripeptide from Streptomyces viridochromogenes Tü494 , 2004, Applied and Environmental Microbiology.

[17]  Shu-Ming Li and Lutz Heide.  Functional analysis of biosynthetic genes of aminocoumarins and production of hybrid antibiotics , 2004 .

[18]  K. Chater,et al.  Production of 8'-halogenated and 8'-unsubstituted novobiocin derivatives in genetically engineered streptomyces coelicolor strains. , 2004, Chemistry & biology.

[19]  W. Gerwick,et al.  Lyngbyatoxin biosynthesis: sequence of biosynthetic gene cluster and identification of a novel aromatic prenyltransferase. , 2004, Journal of the American Chemical Society.

[20]  J. Ravel,et al.  The Biosynthetic Gene Cluster for a Monocyclic β-Lactam Antibiotic, Nocardicin A , 2004 .

[21]  L. Heide,et al.  New aminocoumarin antibiotics from a cloQ-defective mutant of the clorobiocin producer Streptomyces roseochromogenes DS12.976. , 2004, The Journal of antibiotics.

[22]  H. Tan,et al.  Cloning and characterization of sanO, a gene involved in nikkomycin biosynthesis in Streptomyces ansochromogenes , 2003, Letters in applied microbiology.

[23]  Y. Chan,et al.  Deciphering Tuberactinomycin Biosynthesis: Isolation, Sequencing, and Annotation of the Viomycin Biosynthetic Gene Cluster , 2003, Antimicrobial Agents and Chemotherapy.

[24]  L. Heide,et al.  CloN2, a novel acyltransferase involved in the attachment of the pyrrole-2-carboxyl moiety to the deoxysugar of clorobiocin. , 2003, Microbiology.

[25]  Ameriga Lazzarini,et al.  The gene cluster for the biosynthesis of the glycopeptide antibiotic A40926 by nonomuraea species. , 2003, Chemistry & biology.

[26]  T. Luft,et al.  Novobiocin biosynthesis: inactivation of the putative regulatory gene novE and heterologous expression of genes involved in aminocoumarin ring formation , 2003, Archives of Microbiology.

[27]  T. Luft,et al.  Clorobiocin biosynthesis in Streptomyces: identification of the halogenase and generation of structural analogs. , 2003, Chemistry & biology.

[28]  Alex Bateman,et al.  New Knowledge from Old: In silico discovery of novel protein domains in Streptomyces coelicolor , 2003, BMC Microbiology.

[29]  G. Challis,et al.  PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Daniel Bischoff,et al.  Crystal Structure of OxyB, a Cytochrome P450 Implicated in an Oxidative Phenol Coupling Reaction during Vancomycin Biosynthesis* , 2002, The Journal of Biological Chemistry.

[31]  L. Heide,et al.  Molecular cloning and sequence analysis of the clorobiocin biosynthetic gene cluster: new insights into the biosynthesis of aminocoumarin antibiotics. , 2002, Microbiology.

[32]  C. Walsh,et al.  Genetics and Assembly Line Enzymology of Siderophore Biosynthesis in Bacteria , 2002, Microbiology and Molecular Biology Reviews.

[33]  G. Challis,et al.  The vbs genes that direct synthesis of the siderophore vicibactin in Rhizobium leguminosarum: their expression in other genera requires ECF σ factor RpoI , 2002, Molecular microbiology.

[34]  L. Heide,et al.  Cloning and analysis of the simocyclinone biosynthetic gene cluster of Streptomyces antibioticus Tü 6040 , 2002, Archives of Microbiology.

[35]  B. Barrell,et al.  Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2) , 2002, Nature.

[36]  B. Shen,et al.  Cloning and characterization of the bleomycin biosynthetic gene cluster from Streptomyces verticillus ATCC15003. , 2002, Journal of natural products.

[37]  C. Walsh,et al.  Molecular cloning and sequence analysis of the complestatin biosynthetic gene cluster , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[38]  C. Walsh,et al.  Coumarin formation in novobiocin biosynthesis: beta-hydroxylation of the aminoacyl enzyme tyrosyl-S-NovH by a cytochrome P450 NovI. , 2001, Chemistry & biology.

[39]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[40]  E. Bossi,et al.  Teicoplanin biosynthesis genes in Actinoplanes teichomyceticus , 2000, Antonie van Leeuwenhoek.

[41]  M. Doumith,et al.  Analysis of genes involved in 6-deoxyhexose biosynthesis and transfer in Saccharopolyspora erythraea , 2000, Molecular Genetics and Genomics.

[42]  L. Heide,et al.  Identification of the Coumermycin A1Biosynthetic Gene Cluster of Streptomyces rishiriensisDSM 40489 , 2000, Antimicrobial Agents and Chemotherapy.

[43]  B. Wanner,et al.  One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[44]  G. Challis,et al.  Coelichelin, a new peptide siderophore encoded by the Streptomyces coelicolor genome: structure prediction from the sequence of its non-ribosomal peptide synthetase. , 2000, FEMS microbiology letters.

[45]  L. Heide,et al.  Identification of the Novobiocin Biosynthetic Gene Cluster of Streptomyces spheroides NCIB 11891 , 2000, Antimicrobial Agents and Chemotherapy.

[46]  Mark J. Buttner,et al.  Evidence that the Extracytoplasmic Function Sigma Factor ςE Is Required for Normal Cell Wall Structure in Streptomyces coelicolor A3(2) , 1999, Journal of bacteriology.

[47]  C. Walsh,et al.  Identification of a Mycobacterium tuberculosis gene cluster encoding the biosynthetic enzymes for assembly of the virulence-conferring siderophore mycobactin. , 1998, Chemistry & biology.

[48]  B. Barrell,et al.  Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence , 1998, Nature.

[49]  H. Kinashi,et al.  A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome , 1996, Molecular microbiology.

[50]  D. Macneil,et al.  Analysis of Streptomyces avermitilis genes required for avermectin biosynthesis utilizing a novel integration vector. , 1992, Gene.

[51]  D. Hopwood,et al.  CDA is a new chromosomally-determined antibiotic from Streptomyces coelicolor A3(2). , 1983, Journal of general microbiology.

[52]  K. Chater,et al.  Lambda red-mediated genetic manipulation of antibiotic-producing Streptomyces. , 2004, Advances in applied microbiology.

[53]  J. Ravel,et al.  The biosynthetic gene cluster for a monocyclic beta-lactam antibiotic, nocardicin A. , 2004, Chemistry & biology.

[54]  C. Walsh,et al.  Formation of beta-hydroxy histidine in the biosynthesis of nikkomycin antibiotics. , 2002, Chemistry & biology.

[55]  T. Kieser.  Practical streptomyces genetics , 2000 .