Antibiotic production,among the several characteristics of temporally regulated gene expression in Streptomyces, is the major reason for the considerable fundamental and applied scientific interest in these filamentous soil bacteria. Acquisition of new knowledge about the genetic and biochemical basis of antibiotic production will lead to ways to make new antibiotics and to improve their commercial production. It also will provide significant information about prokaryotic molecular biology. Polyether antibiotics, currently used as coccidiostats and bovine growth promotants, are the subject of this proposal for continuing our research on the biosynthesis. The long-range goals are to learn how the microorganisms can make polyethers in such characteristically large amounts by investigating the regulation of antibiotic production, and to learn how they assemble the intricate carbon skeleton of polyether antibiotics by studying the properties of polyether polyketide synthases and the corresponding genes. During this grant period, we will characterize the monensin polyketide synthase and resistance genes by complementation experiments, DNA sequencing, and transcriptional analysis. We will compare the structures of the monensin and salinomycin polyketide synthase genes and predicted gene products. The metabolites accumulated by Mon mutants will be characterized by chemical and spectroscopic analyses to help us identify the properties of monensin production genes. We will clone the valine dehydrogenase genes from S. coelicolor and S. cinnamonensis to study the regulation of vdh expression and the possible connection of valine catabolism to monensin production. finally, we will investigate the genetic basis for the frequent loss and restoration of lasalocid production of S. lasaliensis by following the behavior of the lasalocid polyketide synthase gene in protoplast fusion.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025799-13
Application #
3273305
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1979-01-01
Project End
1993-12-31
Budget Start
1992-03-01
Budget End
1993-12-31
Support Year
13
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Pharmacy
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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

Showing the most recent 10 out of 13 publications