Progress 10/01/06 to 09/30/11
Outputs
OUTPUTS: The outputs completed during this reporting period include conducting experiments to determine how different classes of natural products are biosynthesized by the producing bacteria. The information obtained from these studies was disseminated to the communities of interest through seminars presented at different academic institutions. This information provided fundamentally new information concerning natural product biosynthesis. PARTICIPANTS: Michael Thomas - PI; Elizabeth Felnagle, graduate student in MDTP; Matthew McMahon, graduate student in MDTP; Hyunjun Park, graduate student in MDTP; Donald Drott, graduate student in M.S. program Rosivette Santiago, graduate student in M.S. program; Angela Podevels, scientist; John Barkei, scientist TARGET AUDIENCES: The target audience for these efforts are microbial physiologists, plant biologists, natural product chemists, biochemists. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The results from these studies provided significant new insights into the proteins required for proper natural product assembly. This is best exemplified by the publication of this work in peer-reviewed scientific journals.
Publications
- Park, H. and Thomas, M. G. (2010) Using surrogates to bypass missing catalytic components. Chem. Biol. 17:1045-1046.
- Felnagle, E. A., Barkei, J. J., Park, H., Podevels, A. M., McMahon, M. D., Drott, D. W. and Thomas, M. G. (2010) MbtH-like proteins as integral components of bacterial nonribosomal peptide synthetases. Biochemistry. 49:8815-8817.
- Chan, Y. A. and Thomas, M. G. (2010) Recognition of (2S)-aminomalonyl-acyl carrier protein (ACP) and (2R)-hydroxymalonyl-ACP by acyltransferases in zwittermicin A biosynthesis. Biochemistry. 49:3667-3677.
- Moe, L. A., McMahon, M. D., and M. G. Thomas. (2010) Functional metagenomics as a technique for the discovery of novel enzymes and natural products. In Enzyme Technologies: Metagenomics, Biocatalysis and Biosynthesis. Yeh, W.-K., Yang, H.-C., McCarthy, J. R. Eds. John Wiley & Sons, Inc.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: The outputs completed during this reporting period include conducting experiments to investigate the biosynthetic mechanisms for the production of antibiotics by bacteria. The information obtained from these studies was disseminated by seminars presented at various academic institutions and international meetings. This information provided fundamentally new information concerning nonribosomal peptide and polyketide biosynthesis by bacteria. PARTICIPANTS: Brian M. Kevany, research associate (postdoctoral researcher); Yolande A. Chan, research assistant (graduate student); Elizabeth A. Felnagle, research assistant (graduate student); Matthew D. McMahon, research assistant (graduate student); Hyunjun Park, research assistant (graduate student); Donald Drott, research assistant (graduate student;) John J. Barkei, senior research specialist; Angela M. Podevels, research specialist TARGET AUDIENCES: The target audiences for these efforts are microbial physiologists, biochemists, and natural product chemists PROJECT MODIFICATIONS: None.
Impacts
The results from these studies provided new insights into precursor biosynthesis for the production of nonribosomal peptide and polyketide antibiotics. These data also provided a proof-of-principle that metabolic engineering of antibiotic biosynthetic pathways is a reasonable approach to generating new derivatives of known antibiotics.
Publications
- Berti, A. D. and Thomas, M. G. Analysis of acrhomobactin biosynthesis by Pseudomonas syringae pv. syringae B728a. 2009. J. Bact. 191:4594-4604.
- Helmetag, V, Samel, S. A., Thomas, M. G., Marahiel, M. A., Essen, L-O. Structural basis for the erythro-stereospecificity of the L-arginine oxygenase VioC in viomycin biosynthesis. 2009. FEBS J. 276:3669-3682.
- Chan, YA and Thomas, M. G. Formation and characterization of acyl carrier protein-linked polyketide synthase extender units. Meth. Enzymol. 2009 26:90-114
- Chan, YA, Podevels, AM, Kevany, BM and Thomas, MG Biosynthesis of polyketide synthase extender units. Nat. Prod. Rep. 2009 26:90-114
- Barkei, JJ, Kevany, BM, Felnagle, EA and Thomas,MG. Investigations into viomycin biosynthesis by using heterologous production in Streptomyces lividans. ChemBioChem. 2009 10:366-376.
- Kevany, BM, Rasko, DA, and Thomas, MG Characterization of the complete zwittermicin A biosynthetic gene cluster from Bacillus cereus. Appl Environ Microbiol. 2009 75:1144-1155
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: The outputs completed during this reporting period include conducting experiments to investigate the biosynthetic mechanisms for the production of antibiotics by bacteria. The information obtained from these studies was disseminated by seminars presented at various academic institutions and two international meetings (Gordon Research Conference, Biocatalysis; 7th US-Japan Seminar on the Biosynthesis of Natural Products). This information provided fundamentally new information concerning natural product biosynthesis by bacteria. PARTICIPANTS: Brian M. Kevany, research associate (postdoctoral researcher); Yolande A. Chan, research assistant (graduate student); Elizabeth A. Felnagle, research assistant (graduate student); Matthew D. McMahon, research assistant (graduate student); John J. Barkei, senior research specialist; Angela M. Podevels, research specialist TARGET AUDIENCES: The target audiences for these efforts are microbial physiologists, biochemists, and natural product chemists PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The results from these studies provided new insights into precursor biosynthesis for the production of nonribosomal peptide and polyketide antibiotics. These data also provided a proof-of-principle that metabolic engineering of antibiotic biosynthetic pathways is a reasonable approach to generating new derivatives of known antibiotics.
Publications
- Felnagle, E. A., Jackson, E. E., Chan, Y. A., Podevels, A. M., Berti, A. D., McMahon, M. D., and Thomas, M. G. (2008) Nonribosomal peptide synthetases involved in the production of medically relevant natural products. Molec. Pharmaceutics. 5:191-211.
- Barkei, JJ, Kevany, BM, Felnagle, EA and Thomas, MG. (2008) Investigations into viomycin biosynthesis by using heterologous production in Streptomyces lividans. ChemBioChem. DOI: 10.1002/cbic.200800646
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Progress 01/01/07 to 12/31/07
Outputs
OUTPUTS: The outputs completed during this granting period included conducting and analyzing data that investigated the biosynthesis of antibiotics, specifically those that can target Mycobacterium tuberculosis. A portion of this work was presented at the ASM General Meeting in Toronto, Canada as a means for disseminating the data and conclusions obtained. The output provided unique insights into antibiotic production, specifically those antibiotics that involve nonribosomal peptide synthetase or polyketide synthase enzymology. In the process of this work, preliminary patents were filed. The research conducted was instrumental in the scientific training of four graduate students, two undergraduate students and two technicians.
PARTICIPANTS: Michael G. Thomas (UW-Madison): Principle investigator: performed experiments, interpreted data, mentored students and staff, wrote manuscripts. Elizabeth Felnagle (UW-Madison): graduate student, performed experiments, interpreted data, co-wrote manuscript. Andrew Berti (UW-Madison): graduate student, performed experiments, interpreted data, co-wrote manuscript. Angela Podevels (UW-Madison): Research associate, performed experiments, interpreted data, co-wrote manuscript. Michelle Rondon (UW-Madison): Assistant scientist, performed experiments, interpreted data, co-wrote manuscript. Heidi Crosby (UW-Madison): Graduate student, performed experiments, interpreted data.John Barkei (UW-Madison): Research associate, performed experiments, interpreted data. Michelle Pocholec (Harvard Medical School): graduate student, performed experiments, interpreted data. Jason Sello (Brown University): Assistant professor, performed experiments, interpreted results, co-wrote manuscript.
Christopher Walsh (Harvard Medical School): Professor, interpreted results, co-wrote manuscript.
PROJECT MODIFICATIONS: None
Impacts
The results from these studies provided new insights into the assembly of nonribosomal antibiotics and the resistance to these antibiotics and aminoglycosides. In particular, this work provided new insights into how ribosome modification can lead to cross-resistance to antibiotics. These studies also identified new mechanisms for antibiotic production through heterologous production. The success in heterologous antibiotic production also provided a new means for metabolically engineering biosynthetic pathways to generate new antibiotics.
Publications
- Felnagle, E. A., Rondon, M. R., Berti, A. D., Crosby, H. A., and Thomas, M. G. (2007) Identification of the biosynthetic gene cluster and an additional gene for resistance to the antituberculosis drug capreomycin. Appl. Environ. Microbiol. 73:4162-4170.
- Pacholec, M., Sello, J. K., Walsh, C. T., and Thomas, M. G. (2007) Formation of an aminoacyl-S-enzyme intermediate is a key step in the biosynthesis of chloramphenicol. Org. Biomolec. Chem. 5:1692-1694.
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