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. 1971 Jun;47(6):740–744. doi: 10.1104/pp.47.6.740

Solubilization and Separation of Uridine Diphospho-d-glucose: β-(1 → 4) Glucan and Uridine Diphospho-d-glucose:β-(1 → 3) Glucan Glucosyltransferases from Coleoptiles of Avena sativa1

C M Tsai a, W Z Hassid a
PMCID: PMC396763  PMID: 16657697

Abstract

The particulate glucan synthetase preparation isolated from a homogenate of oat coleoptiles at 4 C lost 65% of its original activity after 1 day when the UDP-d-glucose substrate concentration was 5 × 10−7m to 1.0 × 10−6m. Storage of the particulate enzyme at −20 C or in liquid nitrogen did not prevent the enzyme from losing its activity. Incorporation of 0.5% hovine serum albumin into the medium stabilized the particulate enzyme at 0 C for 6 days and for at least 2 weeks in liquid nitrogen.

When the particulate enzyme was treated with 8% digitonin, 40 to 50% of its activity appeared in the 100,000g supernatant fraction. The particulate and digitonin-solubilized enzyme preparations synthesized both β-(1 → 4) and β-(1 → 3) glucosyl linkages from UDP-d-glucose, but β-(1 → 3) glucan was the main product at 1 × 10−3m UDP-d-glucose substrate. The activity of β-(1 → 4) glucan synthetase was stimulated at least 10-fold in the presence of MgCl2. A separation of β-(1 → 4) and β-(1 → 3) glucan synthetase activities could be achieved at 1 × 10−3m UDP-d-glucose when the digitonin-solubilized enzyme was adsorbed on a hydroxylapatite gel and then eluted with concentrated potassium phosphate buffer. The results indicate that the particulate enzyme contains two enzymes, one responsible for the synthesis of β-(1 → 4) and another β-(1 → 3) linkages in the glucan or glucans synthesized from UDP-d-glucose.

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Selected References

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

  1. BARBER H. A., ELBEIN A. D., HASSID W. Z. THE SYNTHESIS OF CELLULOSE BY ENZYME SYSTEMS FROM HIGHER PLANTS. J Biol Chem. 1964 Dec;239:4056–4061. [PubMed] [Google Scholar]
  2. Batra K. K., Hassid W. Z. Determination of Linkages of beta-d-Glucans from Lupinus albus and Avena sativa by exo-beta-(1 --> 3)-d-Glucanase. Plant Physiol. 1970 Feb;45(2):233–234. doi: 10.1104/pp.45.2.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Batra K. K., Hassid W. Z. Determination of linkages in beta-D-Glucans from Phaseolus aureus by exo-beta-(1 to 3)-D-glucanase. Plant Physiol. 1969 May;44(5):755–758. doi: 10.1104/pp.44.5.755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brummond D. O., Gibbons A. P. Enzymatic cellulose synthesis from UDP-(14C)-glucose by Lupinus albus. Biochem Z. 1965 Aug 6;342(3):308–318. [PubMed] [Google Scholar]
  5. FEINGOLD D. S., NEUFELD E. F., HASSID W. Z. Synthesis of a beta-1, 3-linked glucan by extracts of Phaseolus aureus seedlings. J Biol Chem. 1958 Oct;233(4):783–788. [PubMed] [Google Scholar]
  6. Flowers H. M., Batra K. K., Kemp J., Hassid W. Z. Biosynthesis of Insoluble Glucans From Uridine-Diphosphate-d-Glucose With Enzyme Preparations From Phaseolus aureus and Lupinus albus. Plant Physiol. 1968 Oct;43(10):1703–1709. doi: 10.1104/pp.43.10.1703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Flowers H. M., Batra K. K., Kemp J., Hassid W. Z. Biosynthesis of cellulose in vitro from guanosine diphosphate D-glucose with enzymic preparations from Phaseolus aureus and Lupinus albus. J Biol Chem. 1969 Sep 25;244(18):4969–4974. [PubMed] [Google Scholar]
  8. 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]
  9. Liu T. Y., Hassid W. Z. Solubilization and partial purification of cellulose synthetase from Phaseolus aureus. J Biol Chem. 1970 Apr 25;245(8):1922–1925. [PubMed] [Google Scholar]
  10. Ordin L., Hall M. A. Cellulose synthesis in higher plants from UDP glucose. Plant Physiol. 1968 Mar;43(3):473–476. doi: 10.1104/pp.43.3.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ordin L., Hall M. A. Studies on Cellulose Synthesis by a Cell-free Oat Coleoptile Enzyme System: Inactivation by Airborne Oxidants. Plant Physiol. 1967 Feb;42(2):205–212. doi: 10.1104/pp.42.2.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Villemez C. L., Franz G., Hassid W. Z. Biosynthesis of Alkali Insoluble Polysaccharide from UDP-d-Glucose with Particulate Enzyme Preparations from Phaseolus aureus. Plant Physiol. 1967 Sep;42(9):1219–1223. doi: 10.1104/pp.42.9.1219. [DOI] [PMC free article] [PubMed] [Google Scholar]

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