Abstract
The β-galactosidase (EC 3.2.1.32) of Corynebacterium murisepticum (inducible by lactose and galactose) was purified by successive column chromatography on Sephadex G-200, DEAE-Sephadex A-50 and DEAE-cellulose (DE52). The enzyme was found to be a dimer of identical subunits of molecular mass 100,000 daltons. The K m values of the enzyme for the substrates lactose and o-nitrophenyl-β-d-galactopyranoside (ONPG) are 16.7 mM and 4.4 mM, respectively, indicating, its low affinity for the substrates. The Ouchterlony immunodiffusion method exhibited immunological homogeneity of the enzyme preparation. The catalytic site of the enzyme does not take part in antigen-antibody reaction.
Similar content being viewed by others
References
Beckwith J, Zipser D (1970) The lactose operon. Cold Spring Harbour Laboratory, Cold Spring Harbour, New York
Citti JE, Sandine WE, Elliker PR (1965) β-Galactosidase of Streptococcus lactis. J Bacteriol 89:937–942
Craven GR, Steers E Jr, Anfinsen CB (1965) Purification, composition and molecular weight of the β-galactosidase of Escherichia coli K 12. J Biol Chem 240:2468–2477
Das S, Breunig KD, Hollenberg CP (1985) A positive regulatory element is involved in the induction of the β-galactosidase gene from Kluyveromyces lactis. EMBO J 4:793–798
Dickson RC, Dickson LR, Markin JR (1979) Purification and properties of an inducible β-galactosidase isolated from the yeast Kluyveromyces lactis. J Bacteriol 137:51–61
Efstathiou JD, McKay LL (1976) Plasmids in Streptococcus lactis: Evidence that lactose metabolism and proteinase activity are plasmid linked. Appl Environ Microbiol 32:38–44
Fantes PA, Roberts CF (1973) β-Galactosidase activity and lactose utilisation in Aspergillus nidulans. J Gen Microbiol 77:471–486
Hall BG, Reeve GR (1977) A third β-galactosidase in a strain of Klebsiella that possesses two lac genes. J Bacteriol 132:219–223
Hidalgoc RJ, Goldschmidt R (1977) Induction and general properties of β-galactosidase and β-galactoside permease in Pseudomonas BAL-32. J Bacteriol 129:821–829
Hirata N, Negoro S, Okada H (1984) Molecular basis of isozyme formation of β-galactosidases in Bacillus stearothermophilus: Isolation of two β-galactosidase genes, bgaA and bgaB. J Bacteriol 160:9–14
Lester G, Beyers A (1965) Properties of two β-galactosidases of Neurospora crassa. Biochem Biophys Res Commun 18:725–734
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Macris BJ, Markakis P (1981) Characterisation of extracellular β-d-galactosidase from Fusarium moniliforme grown in whey. Appl Environ Microbiol 41:956–958
Messer A, Dahlqvist A (1966) A one step ultramicro method for the assay of intestinal disaccharidases. Anal Biochem 14:376–392
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbour Laboratory, Cold Spring Harbour, New York
Morse ML, Hill KL, Egan JB, Hengestenberg W (1968) Metabolism of lactose by Staphyloccocus aureus and its genetic basis. J Bacteriol 95:2270–2274
Mozaffar Z, Nakanishi K, Matsuno R, Kamikubo T (1984) Purification and properties of β-galactosidase from Bacillus circulans. Agr Biol Chem 48:3053–3061
Nader de Macias ME, Perdigon G, Pesce de Ruiz Holgado AA, Oliver G (1985) Immunological relationship among β-galactosidase in members of the genus Lactobacillus. Int J Sys Bacteriol 35:103–110
Ouchterlony O (1967) Immunodiffusion and immunoelectrophoresis. In: Weir DM (ed) Handbook of experimental immunology. Blackwell Scientific Publications, Oxford, pp 655–706
Ramana Rao MV, Dutta SM (1981) Purification and properties of β-galactosidase from Streptococcus thermophilus. J Food Sci 46:1419–1423
Snook RJ, McKay LL (1981) Conjugal transfer of lactose fermenting ability among Streptococcus cremoris and Streptococcus lactis strains. Appl Environ Microbiol 42:904–911
Stokes HW, Bettes PW, Hal BG (1985) Sequence of the ebgA gene of Escherichia coli: Comparison with the lac Z gene. Mol Biol Evol 2:469–477
Takenishi S, Watanabe Y, Miwa T, Kobayashi R (1983) Purification and some properties of β-galactosidase from Penicillium multicolor. Agr Biol Chem 47:2533–2540
Ulrich JT, Mc Feter GA, Temple KL (1972) Induction and characterisation of beta-galactosidase in an extreme thermophile. J Bacteriol 110:691–698
Weber K, Pringle JR, Osborn M (1972) Measurement of molecular weight by electrophoresis on SDS-acrylamide gel. Methods in enzymology, vol 26, part C. Academic Press, New York, pp 3–27
Widmar F, Leuba JL (1979) β-Galactosidase from Aspergillus niger: Separation and characterisation of three multiple forms. Eur J Biochem 100:559–567
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Priyolkar, M., Nair, C.K.K. & Pradhan, D.S. Purification and characterisation of an inducible β-galactosidase from Corynebacterium murisepticum . Arch. Microbiol. 151, 49–53 (1988). https://doi.org/10.1007/BF00444668
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00444668