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. 1968 Jun;95(6):2325–2335. doi: 10.1128/jb.95.6.2325-2335.1968

Tryptophan Synthetic Pathway and Its Regulation in Chromobacterium violaceum

Judith Wegman 1, Irving P Crawford 1
PMCID: PMC315168  PMID: 5669904

Abstract

Extracts of Chromobacterium violaceum catalyzed all of the reactions involved in synthesizing tryptophan from chorismic acid. Tryptophan auxotrophs which had lost any of these activities did not produce the characteristic purple pigment, violacein, when grown on a medium in which tryptophan was limiting. Gel filtration of extracts allowed us to estimate molecular weights for the tryptophan enzymes. All of the enzymes appeared to have molecular weights below 100,000. No enzymes were observed to occur in aggregates. The specific activities of the enzymes of the tryptophan pathway did not change when mutants were grown under conditions of limiting or excess tryptophan. The first enzyme in the pathway, anthranilate synthetase, was subject to feedback control by the end product, tryptophan. Tryptophan acted as a noncompetitive inhibitor with respect to glutamine, one of the substrates for anthranilate synthetase, and as a competitive inhibitor of the reaction when chorismate, the other substrate, was varied. The nonlinearity observed in the Lineweaver-Burk plot in the latter case suggests that there may be more than one chorismate-binding site on anthranilate synthetase.

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Selected References

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

  1. ANAGNOSTOPOULOS C., CRAWFORD I. P. Transformation studies on the linkage of markers in the tryptophan pathway in Bacillus subtilis. Proc Natl Acad Sci U S A. 1961 Mar 15;47:378–390. doi: 10.1073/pnas.47.3.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. ATKINSON D. E., HATHAWAY J. A., SMITH E. C. KINETICS OF REGULATORY ENZYMES. KINETIC ORDER OF THE YEAST DIPHOSPHOPYRIDINE NUCLEOTIDE ISOCITRATE DEHYDROGENASE REACTION AND A MODEL FOR THE REACTION. J Biol Chem. 1965 Jun;240:2682–2690. [PubMed] [Google Scholar]
  3. Baker T. I., Crawford I. P. Anthranilate synthetase. Partial purification and some kinetic studies on the enzyme from Escherichia coli. J Biol Chem. 1966 Dec 10;241(23):5577–5584. [PubMed] [Google Scholar]
  4. Bauerle R. H., Margolin P. A multifunctional enzyme complex in the tryptophan pathway of Salmonella typhimurium: comparison of polarity and pseudopolarity mutations. Cold Spring Harb Symp Quant Biol. 1966;31:203–214. doi: 10.1101/sqb.1966.031.01.028. [DOI] [PubMed] [Google Scholar]
  5. Bauerle R. H., Margolin P. The functional organization of the tryptophan gene cluster in Salmonella typhimurium. Proc Natl Acad Sci U S A. 1966 Jul;56(1):111–118. doi: 10.1073/pnas.56.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CRAWFORD I. P., JOHNSON L. M. MUTANTS OF ESCHERICHIA COLI DEFECTIVE IN THE B PROTEIN OF TRYPTOPHAN SYNTHETASE. II. INTRAGENIC POSITION. Genetics. 1964 Feb;49:267–278. doi: 10.1093/genetics/49.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crawford I. P., Gunsalus I. C. Inducibility of tryptophan synthetase in Pseudomonas putida. Proc Natl Acad Sci U S A. 1966 Aug;56(2):717–724. doi: 10.1073/pnas.56.2.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Creighton T. E., Yanofsky C. Association of the alpha and beta-2 subunits of the tryptophan synthetase of Escherichia coli. J Biol Chem. 1966 Feb 25;241(4):980–990. [PubMed] [Google Scholar]
  9. Creighton T. E., Yanofsky C. Indole-3-glycerol phosphate synthetase of Escherichia coli, an enzyme of the tryptophan operon. J Biol Chem. 1966 Oct 25;241(20):4616–4624. [PubMed] [Google Scholar]
  10. DEMOSS J. A. Studies on the mechanism of the tryptophan synthetase reaction. Biochim Biophys Acta. 1962 Aug 13;62:279–293. doi: 10.1016/0006-3002(62)90041-0. [DOI] [PubMed] [Google Scholar]
  11. DEMOSS R. D., EVANS N. R. Incorporation of C14-labeled substrates into violacein. J Bacteriol. 1960 May;79:729–733. doi: 10.1128/jb.79.5.729-733.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. DOY C. H., RIVERA A., Jr, SRINIVASAN P. R. Evidence for the enzymatic synthesis of N-(5'-phosphoribosyl) anthranilic acid, a new intermediate in tryptophan biosynthesis. Biochem Biophys Res Commun. 1961 Jan 25;4:83–88. doi: 10.1016/0006-291x(61)90261-3. [DOI] [PubMed] [Google Scholar]
  13. DeMoss J. A., Wegman J. An enzyme aggregate in the tryptophan pathway of Neurospora crassa. Proc Natl Acad Sci U S A. 1965 Jul;54(1):241–247. doi: 10.1073/pnas.54.1.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Doy C. H., Cooper J. M. Aromatic biosynthesis in yeast. I. The synthesis of tryptophan and the regulation of this pathway. Biochim Biophys Acta. 1966 Oct 31;127(2):302–316. [PubMed] [Google Scholar]
  15. Enatsu T., Crawford I. P. Enzymes of the tryptophan synthetic pathway in Pseudomonas putida. J Bacteriol. 1968 Jan;95(1):107–112. doi: 10.1128/jb.95.1.107-112.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gibson F. Chorismic acid: purification and some chemical and physical studies. Biochem J. 1964 Feb;90(2):256–261. doi: 10.1042/bj0900256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gibson M. I., Gibson F. Preliminary studies on the isolation and metabolism of an intermediate in aromatic biosynthesis: chorismic acid. Biochem J. 1964 Feb;90(2):248–256. doi: 10.1042/bj0900248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hütter R., DeMoss J. A. Enzyme analysis of the tryptophan pathway in Aspergillus nidulans. Genetics. 1967 Feb;55(2):241–247. doi: 10.1093/genetics/55.2.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hütter R., DeMoss J. A. Organization of the tryptophan pathway: a phylogenetic study of the fungi. J Bacteriol. 1967 Dec;94(6):1896–1907. doi: 10.1128/jb.94.6.1896-1907.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ito J., Crawford I. P. Regulation of the enzymes of the tryptophan pathway in Escherichia coli. Genetics. 1965 Dec;52(6):1303–1316. doi: 10.1093/genetics/52.6.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ito J., Yanofsky C. The nature of the anthranilic acid synthetase complex of Escherichia coli. J Biol Chem. 1966 Sep 10;241(17):4112–4114. [PubMed] [Google Scholar]
  22. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  23. MONOD J., CHANGEUX J. P., JACOB F. Allosteric proteins and cellular control systems. J Mol Biol. 1963 Apr;6:306–329. doi: 10.1016/s0022-2836(63)80091-1. [DOI] [PubMed] [Google Scholar]
  24. Nester E. W., Jensen R. A. Control of aromatic acid biosynthesis in Bacillus subtilis: sequenial feedback inhibition. J Bacteriol. 1966 Apr;91(4):1594–1598. doi: 10.1128/jb.91.4.1594-1598.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. RAMAKRISHNAN T., ADELBERG E. A. REGULATORY MECHANISMS IN THE BIOSYNTHESIS OF ISOLEUCINE AND VALINE. II. IDENTIFICATION OF TWO OPERATOR GENES. J Bacteriol. 1965 Mar;89:654–660. doi: 10.1128/jb.89.3.654-660.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SCHWARTZ A. K., BONNER D. M. TRYPTOPHAN SYNTHETASE IN BACILLUS SUBTILIS: EFFECTS OF HIGH POTASSIUM ION CONCENTRATION ON A TWO COMPONENT ENZYME. Biochim Biophys Acta. 1964 Aug 26;89:337–347. doi: 10.1016/0926-6569(64)90223-8. [DOI] [PubMed] [Google Scholar]
  27. SMITH O. H., YANOFSKY C. 1-(o-Carboxyphenylamino)-1-deoxyribulose 5-phosphate, a new intermediate in the biosynthesis of tryptophan. J Biol Chem. 1960 Jul;235:2051–2057. [PubMed] [Google Scholar]
  28. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  29. Wegman J., DeMoss J. A. The enzymatic conversion of anthranilate to indolylglycerol phosphate in Neurospora crassa. J Biol Chem. 1965 Oct;240(10):3781–3788. [PubMed] [Google Scholar]
  30. YANOFSKY C. The enzymatic conversion of anthranilic acid to indole. J Biol Chem. 1956 Nov;223(1):171–184. [PubMed] [Google Scholar]
  31. YANOFSKY C. The tryptophan synthetase system. Bacteriol Rev. 1960 Jun;24(2):221–245. doi: 10.1128/br.24.2.221-245.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]

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