Affiliation:
1. Department of Biology, James Madison University, Harrisonburg, Virginia 22807, USA. Valenthe@vaxl.acs.jmu.cdu
Abstract
The gene encoding the large subunit of the methylmalonyl-coenzyme A (CoA) mutase in Nocardia corallina (mutBNc) was cloned. A 4.3-kbp BamHI fragment containing almost the entire mutBNc was identified by Southern hybridization experiments employing a digoxigenin-labeled probe deduced from mutB of Streptomyces cinnamonensis, mutBNc was interrupted by insertion of a kanamycin resistance gene block (mutB::kan or mutB::neo) and introduced into N. corallina to obtain mutB-negative strains by homologous recombination. Four of sixteen kanamycin-resistant clones occurred via double-crossover events and harbored only the interrupted mutBNc. These exhibited no growth on odd-chain fatty acids in the presence of kanamycin but exhibited wild-type growth on even-chain fatty acids, glucose, and succinate. Whereas the wild type of N. corallina accumulates a copolyester of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) containing more than 60 mol% 3HV from most carbon sources, mutB-negative strains accumulated poly(3HB-co-3HV) containing only 2 to 6 mol% 3HV. Methylmalonyl-CoA mutase activity was not found in these clones. Therefore, this study provides strong evidence that the majority of 3HV units in poly(3HB-co-3HV) accumulated by N. corallina are synthesized via the methylmalonyl-CoA pathway.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Reference32 articles.
1. Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates;Anderson A. J.;Microbiol. Rev.,1990
2. Anderson A. J. G. W. Haywood D. R. Williams and E. A. Dawes. 1990. The production of polyhydroxyalkanoates from unrelated carbon sources p. 119-129. In E. A. Dawes (ed.) Novel biodegradable microbial polymers. Kluwer Academic Publishers Dordrecht The Netherlands.
3. Ausubel F. M. R. Brent R. E. Kingston D. D. Moore J. G. Seidman J. A. Smith and K. Struhl (ed.). 1987. Current protocols in molecular biology. Greene Publishing Associates and John Wiley & Sons New York.
4. Bachmann B. J. 1987. Linkage map of Escherichia coli K-12 edition 7 p. 807-876. In F. C. Neidhardt J. L. Ingraham K. B. Low B. Magasanik M. Schaechter and H. E. Umbarger (ed.) Escherichia coli and Salmonella typhimurium: cellular and molecular biology vol. 2. American Society for Microbiology Washington D.C.
5. Cloning, sequencing, and expression of the gene encoding methylmalonyl-coenzyme A mutase from Streptomyces cinnamonensis;Birch A.;J. Bacteriol.,1993
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