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. 1984 Dec;76(4):885–888. doi: 10.1104/pp.76.4.885

Localization and Substrate Specificity of Glycosidases in Vacuoles of Nicotiana rustica

James A Saunders 1,2, June M Gillespie 1,2
PMCID: PMC1064399  PMID: 16663964

Abstract

Vacuoles isolated from Nicotiana rustica var brasilia have been shown to contain significant levels of glycosidase activity when assayed using p-nitrophenyl-glycosides as substrates. The substrate specificity for the glycosidases in the vacuolar fraction closely paralleled that found in the protoplasts, and the leaf tissue from which the vacuoles were isolated. The substrate specificity of the vacuolar enzyme(s) was different from glycosidic activity found in the commercial digestive enzyme preparations used to isolate the protoplasts from leaf tissue. It was demonstrated that 70 to 90% of the glycosidases that were found in the protoplasts appeared to be localized within the vacuole, when the p-nitrophenyl substrates α- and β-;d-galactose, β-d-glucose, and α-d-mannose were used. Neither the vacuolar nor the protoplast enzymes were active towards the naturally occurring phenolic glycoside, rutin. α-Mannosidase appears to be a valuable marker enzyme for vacuoles isolated from mesophyll leaf cells of tobacco.

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Selected References

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

  1. Boller T., Kende H. Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 1979 Jun;63(6):1123–1132. doi: 10.1104/pp.63.6.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Butcher H. C., Wagner G. J., Siegelman H. W. Localization of Acid hydrolases in protoplasts: examination of the proposed lysosomal function of the mature vacuole. Plant Physiol. 1977 Jun;59(6):1098–1103. doi: 10.1104/pp.59.6.1098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kojima M., Poulton J. E., Thayer S. S., Conn E. E. Tissue Distributions of Dhurrin and of Enzymes Involved in Its Metabolism in Leaves of Sorghum bicolor. Plant Physiol. 1979 Jun;63(6):1022–1028. doi: 10.1104/pp.63.6.1022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Oba K., Conn E. E., Canut H., Boudet A. M. Subcellular Localization of 2-(beta-d-Glucosyloxy)-Cinnamic Acids and the Related beta-glucosidase in Leaves of Melilotus alba Desr. Plant Physiol. 1981 Dec;68(6):1359–1363. doi: 10.1104/pp.68.6.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Saunders J. A., Conn E. E., Lin C. H., Shimada M. Localization of Cinnamic Acid 4-Monooxygenase and the Membrane-bound Enzyme System for Dhurrin Biosynthesis in Sorghum Seedlings. Plant Physiol. 1977 Oct;60(4):629–634. doi: 10.1104/pp.60.4.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Saunders J. A., Conn E. E. Presence of the cyanogenic glucoside dhurrin in isolated vacuoles from sorghum. Plant Physiol. 1978 Feb;61(2):154–157. doi: 10.1104/pp.61.2.154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Saunders J. A. Investigations of vacuoles isolated from tobacco: I. Quantitation of nicotine. Plant Physiol. 1979 Jul;64(1):74–78. doi: 10.1104/pp.64.1.74. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Thayer S. S., Conn E. E. Subcellular Localization of Dhurrin beta-Glucosidase and Hydroxynitrile Lyase in the Mesophyll Cells of Sorghum Leaf Blades. Plant Physiol. 1981 Apr;67(4):617–622. doi: 10.1104/pp.67.4.617. [DOI] [PMC free article] [PubMed] [Google Scholar]

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