Abstract

The focus of the research to be pursued in the next three-year period will be an attempt to establish the rudiments of the biochemical genetics fo polyether antiobiotic biosynthesis by Streptomyces spp. We will study the biosynthesis of three polyether antibiotics: lasalocid A (S. lasaliensis), monensin A (S. cinnamonensis), and salinomycin (S. albus) by biochemical, mutational, and gene cloning experiments. In the comtinuation of our lasalocid A study, we will develop a collection of non-producing (Las-) mutants and identify compunds accumulated by them in fermentation broth. The results of cosynthesis and biotransformation experiments will be used to obtain a biochemical complementation map of the lasalocid A biosynthetic pathway. The gene for lasalocid A self-resistance will be cloned from S. lasaliensis. Then two approached to cloning las genes will be investigated using the Las- mutants and the las resistance gene. The las genes cloned will be mapped by restriction endonuclease digestion, and attempts will be made to map las genes genetically. Mutational cloning of las genes will be used to define the number and size of the transcriptional units of the las gene. The results of this work will be interpreted in relation to a hypothesis for the genetic organization of the putative polyether synthetase (PES) multifunctional protein system. A study of the regulation of lasalocid A production will be initiated by focusing on the role of valine catabolism in the process. Analogous studies will be initiated with the Streptomyces producing monesin A and salinomycin as part of our long-range research goal to create hybrid polyether antibiotics by artificial translocation of DNA encoding the PEs gene.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025799-09
Application #
3273302
Study Section
(SSS)
Project Start
1979-01-01
Project End
1989-02-28
Budget Start
1987-12-01
Budget End
1989-02-28
Support Year
9
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Type
Schools of Pharmacy
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Gallo, M A; Ward, J; Hutchinson, C R (1996) The dnrM gene in Streptomyces peucetius contains a naturally occurring frameshift mutation that is suppressed by another locus outside of the daunorubicin-production gene cluster. Microbiology 142 ( Pt 2):269-75
Tang, L; Zhang, Y X; Hutchinson, C R (1994) The genetic basis of precursor supply for the biosynthesis of macrolide and polyether antibiotics. Ann N Y Acad Sci 721:105-16
Hutchinson, C R (1994) Drug synthesis by genetically engineered microorganisms. Biotechnology (N Y) 12:375-80
Tang, L; Hutchinson, C R (1993) Sequence, transcriptional, and functional analyses of the valine (branched-chain amino acid) dehydrogenase gene of Streptomyces coelicolor. J Bacteriol 175:4176-85
Hutchinson, C R; Decker, H; Madduri, K et al. (1993) Genetic control of polyketide biosynthesis in the genus Streptomyces. Antonie Van Leeuwenhoek 64:165-76
Navarrete, R M; Vara, J A; Hutchinson, C R (1990) Purification of an inducible L-valine dehydrogenase of Streptomyces coelicolor A3(2). J Gen Microbiol 136:273-81
Hutchinson, C R; Borell, C W; Otten, S L et al. (1989) Drug discovery and development through the genetic engineering of antibiotic-producing microorganisms. J Med Chem 32:929-37
Kinashi, H; Otten, S L; Duncan, J S et al. (1988) Frequent loss and restoration of antibiotic production by Streptomyces lasaliensis. J Antibiot (Tokyo) 41:624-37
Gonzalez de la Parra, M; Hutchinson, C R (1987) Macrolide biosynthesis: stereochemistry of the hydroxylation of brefeldin C. J Antibiot (Tokyo) 40:1170-4
Sherman, M M; Hutchinson, C R (1987) Biosynthesis of lasalocid A: biochemical mechanism for assembly of the carbon framework. Biochemistry 26:438-45

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