This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Malmberg, L H Articles by Sherman, D H Search for Related Content PubMed PubMed Citation Articles by Malmberg, L H Articles by Sherman, D H

 Previous Article  |  Next Article 

J Bacteriol. 1993 November; 175(21): 6916-6924

research-article

Precursor flux control through targeted chromosomal insertion of the lysine epsilon-aminotransferase (lat) gene in cephamycin C biosynthesis.

Department of Chemical Engineering and Material Science, University of Minnesota, Minneapolis 55455.

ABSTRACT

Targeted gene insertion methodology was used to study the effect of perturbing alpha-aminoadipic acid precursor flux on the overall production rate of beta-lactam biosynthesis in Streptomyces clavuligerus. A high-copy-number plasmid containing the lysine epsilon-aminotransferase gene (lat) was constructed and used to transform S. clavuligerus. The resulting recombinant strain (LHM100) contained an additional complete copy of lat located adjacent to the corresponding wild-type gene in the chromosome. Biological activity and production levels of beta-lactam antibiotics were two to five times greater than in wild-type S. clavuligerus. Although levels of lysine epsilon-aminotransferase were elevated fourfold in LHM100, the level of ACV synthetase, whose gene is located just downstream of lat, remained unchanged. These data strongly support the notion that direct perturbation of alpha-aminoadipic acid precursor flux resulted in increased antibiotic production. This strategy represents a successful application of metabolic engineering based on theoretical predictions of precursor flux in a secondary metabolic pathway.

This article has been cited by other articles:

• Reddy, M. C.M., Gokulan, K., Jacobs, W. R. Jr, Ioerger, T. R., Sacchettini, J. C. (2008). Crystal structure of Mycobacterium tuberculosis LrpA, a leucine-responsive global regulator associated with starvation response. Protein Sci. 17: 159-170 [Abstract] [Full Text]  
• Alexander, D. C., Anders, C. L., Lee, L., Jensen, S. E. (2007). pcd Mutants of Streptomyces clavuligerus Still Produce Cephamycin C. J. Bacteriol. 189: 5867-5874 [Abstract] [Full Text]  
• Hernando-Rico, V., Martin, J. F., Santamarta, I., Liras, P. (2001). Structure of the ask-asd operon and formation of aspartokinase subunits in the cephamycin producer 'Amycolatopsis lactamdurans'. Microbiology 147: 1547-1555 [Abstract] [Full Text]  
• Paradkar, A. S., Mosher, R. H., Anders, C., Griffin, A., Griffin, J., Hughes, C., Greaves, P., Barton, B., Jensen, S. E. (2001). Applications of Gene Replacement Technology to Streptomyces clavuligerus Strain Development for Clavulanic Acid Production. Appl. Environ. Microbiol. 67: 2292-2297 [Abstract] [Full Text]  
• Khetan, A., Hu, W.-S., Sherman, D. H. (2000). Heterogeneous distribution of lysine 6-aminotransferase during cephamycin C biosynthesis in Streptomyces clavuligerus demonstrated using green fluorescent protein as a reporter. Microbiology 146: 1869-1880 [Abstract] [Full Text]  
• Li, R., Khaleeli, N., Townsend, C. A. (2000). Expansion of the Clavulanic Acid Gene Cluster: Identification and In Vivo Functional Analysis of Three New Genes Required for Biosynthesis of Clavulanic Acid by Streptomyces clavuligerus. J. Bacteriol. 182: 4087-4095 [Abstract] [Full Text]