Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1989 Jun;90(2):567–574. doi: 10.1104/pp.90.2.567

A β-Galactosidase from Radish (Raphanus sativus L.) Seeds

Masayuki Sekimata 1,2, Kiyoshi Ogura 1,2, Yoichi Tsumuraya 1,2, Yohichi Hashimoto 1,2, Shigeru Yamamoto 1,2
PMCID: PMC1061762  PMID: 16666809

Abstract

A basic β-galactosidase (β-Galase) has been purified 281-fold from imbibed radish (Raphanus sativus L.) seeds by conventional purification procedures. The purified enzyme is an electrophoretically homogeneous protein consisting of a single polypeptide with an apparent molecular mass of 45 kilodaltons and pl values of 8.6 to 8.8. The enzyme was maximally active at pH 4.0 on p-nitrophenyl β-d-galactoside and β-1,3-linked galactobiose. The enzyme activity was inhibited strongly by Hg2+ and 4-chloromercuribenzoate. d-Galactono-(1→4)-lactone and d-galactal acted as potent competitive inhibitors. Using galactooligosaccharides differing in the types of linkage as the substrates, it was demonstrated that radish seed β-Galase specifically split off β-1,3- and β-1,6-linked d-galactosyl residues from the nonreducing ends, and their rates of hydrolysis increased with increasing chain lengths. Radish seed and leaf arabino-3,6-galactan-proteins were resistant to the β-galase alone but could be partially degraded by the enzyme after the treatment with a fungal α-l-arabinofuranosidase leaving some oligosaccharides consisting of d-galactose, uronic acid, l-arabinose, and other minor sugar components besides d-galactose as the main product.

Full text

PDF
568

Images in this article

571Figure 2
on p.571
573Figure 4
on p.573

Selected References

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

  1. Arakawa M., Ogata S., Muramatsu T., Kobata A. Beta-galactosidases from jack bean meal and almond emulsin. Application for the enzymatic distinction of Galbeta1 leads to 4GlcNAc and Galbeta1 leads to 3GlcNAc linkages. J Biochem. 1974 Apr;75(4):707–714. doi: 10.1093/oxfordjournals.jbchem.a130443. [DOI] [PubMed] [Google Scholar]
  2. Bhalla P. L., Dalling M. J. Characteristics of a beta-Galactosidase Associated with the Stroma of Chloroplasts Prepared from Mesophyll Protoplasts of the Primary Leaf of Wheat. Plant Physiol. 1984 Sep;76(1):92–95. doi: 10.1104/pp.76.1.92. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradbury J. H., Jenkins G. A. Determination of the structures of trisaccharides by 13C-n.m.r. spectroscopy. Carbohydr Res. 1984 Mar 1;126(1):125–156. doi: 10.1016/0008-6215(84)85131-9. [DOI] [PubMed] [Google Scholar]
  4. Dey P. M., Del Campillo E. Biochemistry of the multiple forms of glycosidases in plants. Adv Enzymol Relat Areas Mol Biol. 1984;56:141–249. doi: 10.1002/9780470123027.ch3. [DOI] [PubMed] [Google Scholar]
  5. Edwards M., Bowman Y. J., Dea I. C., Reid J. S. A beta-D-galactosidase from nasturtium (Tropaeolum majus L.) cotyledons. Purification, properties, and demonstration that xyloglucan is the natural substrate. J Biol Chem. 1988 Mar 25;263(9):4333–4337. [PubMed] [Google Scholar]
  6. Edwards M., Dea I. C., Bulpin P. V., Reid J. S. Purification and properties of a novel xyloglucan-specific endo-(1----4)-beta-D-glucanase from germinated nasturtium seeds (Tropaeolum majus L.). J Biol Chem. 1986 Jul 15;261(20):9489–9494. [PubMed] [Google Scholar]
  7. HUGHES R. C., JEANLOZ R. W. THE EXTRACELLULAR GLYCOSIDASES OF DIPLOCOCCUS PNEUMONIAE. I. PURIFICATION AND PROPERTIES OF A NEURAMINIDASE AND A BETA-GALACTOSIDASE. ACTION ON THE ALPHA-1-ACID GLYCOPROTEIN OF HUMAN PLASMA. Biochemistry. 1964 Oct;3:1535–1543. doi: 10.1021/bi00898a025. [DOI] [PubMed] [Google Scholar]
  8. Holden D. W., Rohringer R. Peroxidases and glycosidases in intercellular fluids from noninoculated and rust-affected wheat leaves : isozyme assay on nitrocellulose blots from two-dimensional gels. Plant Physiol. 1985 Nov;79(3):820–824. doi: 10.1104/pp.79.3.820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Labavitch J. M., Freeman L. E., Albersheim P. Structure of plant cell walls. Purification and characterization of a beta-1,4-galactanase which degrades a structural component of the primary cell walls of dicots. J Biol Chem. 1976 Oct 10;251(19):5904–5910. [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Li S. C., Mazzotta M. Y., Chien S. F., Li Y. T. Isolation and characterization of jack bean beta-galactosidase. J Biol Chem. 1975 Sep 10;250(17):6786–6791. [PubMed] [Google Scholar]
  12. Matheson N. K., Saini H. S. alpha-L-Arabinofuranosidases and beta-D-galactosidases in germinating-lupin cotyledons. Carbohydr Res. 1977 Aug;57:103–116. doi: 10.1016/s0008-6215(00)81924-2. [DOI] [PubMed] [Google Scholar]
  13. Pressey R. beta-Galactosidases in Ripening Tomatoes. Plant Physiol. 1983 Jan;71(1):132–135. doi: 10.1104/pp.71.1.132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
  15. Saunders J. A., Gillespie J. M. Localization and Substrate Specificity of Glycosidases in Vacuoles of Nicotiana rustica. Plant Physiol. 1984 Dec;76(4):885–888. doi: 10.1104/pp.76.4.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Singh M. B., Knox R. B. Quantitative cytochemistry of beta-galactosidase in normal and enzyme deficient (gal) pollen of Brassica campestris: application of the indigogenic method. Histochem J. 1984 Dec;16(12):1273–1296. doi: 10.1007/BF01003726. [DOI] [PubMed] [Google Scholar]
  17. Tsumuraya Y., Ogura K., Hashimoto Y., Mukoyama H., Yamamoto S. Arabinogalactan-Proteins from Primary and Mature Roots of Radish (Raphanus sativus L.). Plant Physiol. 1988 Jan;86(1):155–160. doi: 10.1104/pp.86.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Uesaka E., Sato M., Raiju M., Kaji A. Alpha-l-arabinofuranosidase from Rhodotorula flava. J Bacteriol. 1978 Mar;133(3):1073–1077. doi: 10.1128/jb.133.3.1073-1077.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. WALLENFELS K., LEHMANN J., MALHOTRA O. P. [Research on lactose-splitting enzymes. VII. The specificity of beta-galactosidase from E. coli ML 309]. Biochem Z. 1960;333:209–225. [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES