Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Dec;61(12):4477–4479. doi: 10.1128/aem.61.12.4477-4479.1995

Mutations in the trpD gene of Corynebacterium glutamicum confer 5-methyltryptophan resistance by encoding a feedback-resistant anthranilate phosphoribosyltransferase.

J P O'Gara 1, L K Dunican 1
PMCID: PMC167758  PMID: 8534114

Abstract

The trpD gene from tryptophan-hyperproducing Corynebacterium glutamicum ATCC 21850 was isolated on the basis of its ability to confer resistance to 5-methyltryptophan on wild-type C. glutamicum AS019. Comparative sequence analysis of the genes from the wild-type AS019 and ATCC 21850 trpD genes revealed two amino acid substitutions at the protein level. Further analysis demonstrated that the trpD gene product from ATCC 21850, anthranilate phosphoribosyltransferase, was more resistant to feedback inhibition by either tryptophan or 5-methyltryptophan than its wild-type counterpart. It is proposed that phosphoribosyltransferase insensitivity to tryptophan in ATCC 21850 contributes to an elevated level of tryptophan biosynthesis.

Full Text

The Full Text of this article is available as a PDF (182.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Chou P. Y., Fasman G. D. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. doi: 10.1002/9780470122921.ch2. [DOI] [PubMed] [Google Scholar]
  3. Heery D. M., Dunican L. K. Cloning of the trp gene cluster from a tryptophan-hyperproducing strain of Corynebacterium glutamicum: identification of a mutation in the trp leader sequence. Appl Environ Microbiol. 1993 Mar;59(3):791–799. doi: 10.1128/aem.59.3.791-799.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
  5. Liebl W., Ehrmann M., Ludwig W., Schleifer K. H. Transfer of Brevibacterium divaricatum DSM 20297T, "Brevibacterium flavum" DSM 20411, "Brevibacterium lactofermentum" DSM 20412 and DSM 1412, and Corynebacterium glutamicum and their distinction by rRNA gene restriction patterns. Int J Syst Bacteriol. 1991 Apr;41(2):255–260. doi: 10.1099/00207713-41-2-255. [DOI] [PubMed] [Google Scholar]
  6. O'Gara J. P., Dunican L. K. Direct evidence for a constitutive internal promoter in the tryptophan operon of Corynebacterium glutamicum. Biochem Biophys Res Commun. 1994 Sep 15;203(2):820–827. doi: 10.1006/bbrc.1994.2256. [DOI] [PubMed] [Google Scholar]
  7. Pabst M. J., Kuhn J. C., Somerville R. L. Feedback regulation in the anthranilate aggregate from wild type and mutant strains of Escherichia coli. J Biol Chem. 1973 Feb 10;248(3):901–914. [PubMed] [Google Scholar]
  8. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Spratt B. G., Hedge P. J., te Heesen S., Edelman A., Broome-Smith J. K. Kanamycin-resistant vectors that are analogues of plasmids pUC8, pUC9, pEMBL8 and pEMBL9. Gene. 1986;41(2-3):337–342. doi: 10.1016/0378-1119(86)90117-4. [DOI] [PubMed] [Google Scholar]
  10. Sugimoto S., Shiio I. Enzymes of the tryptophan synthetic pathway in Brevibacterium flavum. J Biochem. 1977 Apr;81(4):823–833. doi: 10.1093/oxfordjournals.jbchem.a131546. [DOI] [PubMed] [Google Scholar]
  11. Yoshihama M., Higashiro K., Rao E. A., Akedo M., Shanabruch W. G., Follettie M. T., Walker G. C., Sinskey A. J. Cloning vector system for Corynebacterium glutamicum. J Bacteriol. 1985 May;162(2):591–597. doi: 10.1128/jb.162.2.591-597.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES