Skip to main content
Log in

Essential groups and stability of α-glucosidase ofPenicillium notatum

  • Food Microbiology
  • Original Articles
  • Published:
Annals of Microbiology Aims and scope Submit manuscript

Abstract

α-Glucosidase (α-D-glucoside glucohydrolase, EC 3.2.1.20) was isolated fromPenicillium notatum. The enzyme was induced by gibberellic acid (GA3). The GA3-mediated increase in the enzyme activity was repressed in presence of abscisic acid, cycloheximide and the antibiotics chloramphenicol, cordycepin, and rifampicin which are inhibitors of protein synthesis. α-Glucosidase was purified 440-fold with of 27.8-fold of purification. The enzyme was immobilized using chitosan gel. The optimal pH values were 6.5 and 7.5 for free and the immobilized enzymes, respectively. The optimal temperatures were 50 and 65 °C for free and immobilized enzymes, respectively. The enzyme hydrolyzed maltose, sucrose, isomaltose, maltotriose but not starch, amylopectin and amylose. Trehalose and glycerol protected the free and immobilized enzymes against inactivation at 70 °C, however trehalose was the better protector. Phytate protected the free enzyme against heat inactivation at both 65 and 70 °C. 2,4,6 Trinitrobenzenesulfonic acid, butanedione and diethylpyrocarbonate inactivated the enzyme and suggest that, lysyl, arginyl and histidyl groups are taking part in enzyme catalysis. The inactivation by the three compounds was protected by the substratepara-nitrophenyl-α-D-glucopyranoside. Treatment of the enzyme with 1-ethyl-3(3-dimethyl aminopropyl)-carbodiimide,p-chloromercuribenzoate, N-bromosuccinimide, N-acetlyimidazole revealing the involving of carboxyl, sulfhydryl, trptophenyl and tyrosyl groups, respectively in the catalysis of α-glucosidase. EDTA,o-phenanthroline, dipyridyl and 8-quinolinol inhibited the enzyme activity and the inhibition was higher in case of free enzyme compared with immobilized enzyme.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bihzad M.A., El-Shora H.M. (1996). Phosphoenolpyruvate carboxylase fromRumex dentatus a C3-plant. J. Plant Physiol., 149: 669–676.

    CAS  Google Scholar 

  • Brazdova B., Tan N.S., Samoshina N.M., Samoshin V.V. (2009). Novel-easily accessible. glucosidase inhibitors: 4-hydroxy-5-alkoxy-1,2 cyclohexanedicarboxylic acids. Carb. Res., 344: 311–321.

    Article  CAS  Google Scholar 

  • Butler S.L., Falke J.J. (1996). Effects of protein stabilizing agents on thermal backbone motions a disulfide trapping study, Biochemistry, 35: 10595–10600.

    Article  CAS  Google Scholar 

  • Cheetham P.S.J. (1985). Principles of industrial biocatalysis and bioprocessing. In: Wiseman, Ed., Handbook of Enzyme Biotechnology, Ellis Horwood Ltd., London, pp. 83–233.

    Google Scholar 

  • El-Shora H.M. (1993). Biochemical studies on phosphoenolpyruvate carboxylase extracted from tubers ofSolanum tuberosum. Bull. Fac. Zagazig. Univ. Egypt, 15 (2): 95–121.

    Google Scholar 

  • El-Shora H.M. (2001). Effect of growth regulators and group modifier on NADH-glutamate synthase of marrow cotyledons. On line J. Biol. Sci., 1 (7): 597–602.

    Article  Google Scholar 

  • El-Shora H.M. (2002). Properties of phenylalanine ammonialyase from marrow cotyledons. Plant Sci., 162: 1–7.

    Article  CAS  Google Scholar 

  • El-Shora H.M., Metwally M. (2008). Effect of phytohormones and group selective reagents on acid phosphatase fromCladosporium cladosporioides. Asian J. Biotechnol., 1 (1): 1–11.

    Google Scholar 

  • Ezeji T., Bahl H. (2006). Purification, characterization, and synergistic action of phytate-resistant α-amylase and α-glucosidase fromGeobacillus thermodenitrificans HRO10. J. Biotechnol., 125: 27–38.

    Article  CAS  PubMed  Google Scholar 

  • Faridmoayer A., Scaman C.H. (2005). Binding residues and catalytic domain of solubleSaccharomyces cerevisiae processing alpha-glucosidase I. Glycobiology, 15 (12): 1341–1348.

    Article  CAS  PubMed  Google Scholar 

  • Giannesi G.C., Polizeli M., Terenzi H., Jorge J. (2006). A novel α-glucosidase fromChaetomium thermophilum var.coprophilum that converts maltose into trehalose: Purification and partial characterization of the enzyme. Process Biochem., 41: 1729–1735.

    Article  CAS  Google Scholar 

  • Gote M.M., Khan M.I., Khire J.M. (2007). Active site directed chemical modification of α-glucosidase fromBacillus stearothemophillus (NCIM 5146): Involvement of lysine, tryptophane and carboxylate residues in catalytic site. Enzyme Microb. Technol., 40: 1312–1320.

    Article  CAS  Google Scholar 

  • Graf E. (1983). Calcium binding to phytic acid. J. Agric. Food, 31: 851–855.

    Article  CAS  Google Scholar 

  • Gupta M.N., Mattiasson B. (1992). Unique applications of immobilized proteins in bioanalytical systems. In: Suelter C.H., Kricka L, Eds, Bioanalytical Applications of Enzymes, Vol. 36, John Wiley & Sons Inc., New York, pp. 1–34.

    Google Scholar 

  • Kerovuo J., Lauraeous M., Nurminen P., Kalkkinen N., Apajalahti J. (1998). Isolation, characterization, molecular gene cloning, and sequencing of a novel phytaseBacillus subtilis. Appl. Environ. Microbiol., 64: 2079–2085.

    CAS  PubMed  Google Scholar 

  • Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacterio phage. Nature 227, 680–685.

    Article  CAS  PubMed  Google Scholar 

  • Leontievsky A.A., Myasoedova N.M., Baskumov B.P., Golovleva L.A., Bucke C., Evans C.S. (2001). Transformation of 2,4,6-trichlorophenol by free and immobilized fungal laccase. Appl. Microbiol. Biotechnol., 57: 85–91.

    Article  CAS  PubMed  Google Scholar 

  • Lin T.Y., Timashe S.N. (1996). On the role of surface tension in the stabilization of globular proteins, Protein Sci., 5: 372–381.

    Article  CAS  PubMed  Google Scholar 

  • Mala S., Kralova B. (2000). Heterooligosacchride synthesis catalyzed by α-glucosidase fromBacillus stearothemophillus. J. Mol. Catalysis B, 10: 617–621.

    Article  CAS  Google Scholar 

  • Miroliaei M., Nemat-Gorgani M. (2001). Sugars protect native and apo yeast alcohol dehydrogenase against irreversible thermoinactivation. Enzyme Microb. Technol., 29: 54–59.

    Article  Google Scholar 

  • Roig M.G., Kennedy F.J. (1992). Perspectives for chemical modification of enzymes. Crit. Rev. Biotechnol., 12: 391–412.

    Article  CAS  Google Scholar 

  • Rowe G.E., Margaritis A. (2004). Enzyme kinetic properties of α-glucosidase inBacillus thuringiensis. Biochem. Eng. J., 17: 121–128.

    Article  CAS  Google Scholar 

  • Sedmak J.J., Grossberg S.E. (1977). A rapid, sensitive and versatile assay for protein using Coomassie Brilliant Blue G-250. Anal. Biochem., 79: 544–552.

    Article  CAS  PubMed  Google Scholar 

  • Spector T. (1978). Refinement of the Coomassie blue method of protein quantitation. Anal. Biochem., 86: 142–146.

    Article  CAS  PubMed  Google Scholar 

  • Tsou C.L. (1998). The role of active site flexibility in enzyme catalysis. Biochemistry (Mosc.), 63: 253–300.

    CAS  Google Scholar 

  • Zdzieb A., Synowiecki J. (2002). New source of the thermostable α-glucosidase suitable for single step starch processing. Food Chem., 79: 485–491.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hamed M. El-Shora.

Rights and permissions

Reprints and permissions

About this article

Cite this article

El-Shora, H.M., Metwally, M.A. & Khlaf, S.A. Essential groups and stability of α-glucosidase ofPenicillium notatum . Ann. Microbiol. 59, 285–291 (2009). https://doi.org/10.1007/BF03178330

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03178330

Key words

Navigation