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. 1973 Jun;51(6):1055–1060. doi: 10.1104/pp.51.6.1055

Partial Purification and Properties of d-Glucosamine 6-Phosphate N-Acetyltransferase from Phaseolus aureus1

Mahmood Vessal a,2, W Z Hassid a
PMCID: PMC366403  PMID: 16658464

Abstract

d-Glucosamine-6-P N-acetyltransferase (EC 2.3.1.4) from mung bean seeds (Phaseolus aureus) was purified 313-fold by protamine sulfate and isoelectric precipitation, ammonium sulfate and acetone fractionation, and CM Sephadex column chromatography. The partially purified enzyme was highly specific for d-glucosamine-6-P. Neither d-glucosamine nor d-galactosamine could replace this substrate. The partially purified enzyme preparation was inhibited up to 50% by 2 × 10−2m EDTA, indicating the requirement of a divalent cation. Among divalent metal ions tested, Mg2+ was required for maximum activity of the enzyme. Mn2+ and Zn2+ were inhibitory, while Co2+ had no effect on the enzyme activity. The pH optimum of the enzyme in sodium acetate and sodium citrate buffers was found to be 5.2. The effect of Mg2+ on the enzyme in sodium acetate and sodium citrate buffers was particularly noticeable in the range of optimum pH. Km values of 15.1 × 10−4m and 7.1 × 10−4m were obtained for d-glucosamine-6-P and acetyl CoA, respectively. The enzyme was completely inhibited by 1 × 10−4mp-hydroxymercuribenzoate, and this inhibition was partially reversed by l-cysteine; indicating the presence of sulfhydryl groups at or near the active site of the enzyme.

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Selected References

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

  1. BROWN D. H. The D-glucosamine-6-phosphate N-acetylase of yeast. Biochim Biophys Acta. 1955 Mar;16(3):429–431. doi: 10.1016/0006-3002(55)90249-3. [DOI] [PubMed] [Google Scholar]
  2. DAVIDSON E. A., BLUMENTHAL H. J., ROSEMAN S. Glucosamine metabolism. II. Studies on glucosamine 6-phosphate N-acetylase. J Biol Chem. 1957 May;226(1):125–133. [PubMed] [Google Scholar]
  3. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  4. LELOIR L. F., CARDINI C. E. The biosynthesis of glucosamine. Biochim Biophys Acta. 1953 Sep-Oct;12(1-2):15–22. doi: 10.1016/0006-3002(53)90119-x. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. REISSIG J. L., STORMINGER J. L., LELOIR L. F. A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem. 1955 Dec;217(2):959–966. [PubMed] [Google Scholar]
  7. ROE J. H., PAPADOPOULOS N. M. The determination of fructose-6-phosphate and fructose-1,6-diphosphate. J Biol Chem. 1954 Oct;210(2):703–707. [PubMed] [Google Scholar]
  8. Vessal M., Hassid W. Z. Partial Purification and Properties of l-Glutamine d-Fructose 6-Phosphate Amidotransferase from Phaseolus aureus. Plant Physiol. 1972 Jun;49(6):977–981. doi: 10.1104/pp.49.6.977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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