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A new glucose oxidase from Aspergillus niger: characterization and regulation studies of enzyme and gene

  • Applied Genetics and Regulation
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Abstract

A new glucose oxidase from Aspergillus niger was isolated and characterized. The enzyme showed different kinetic and stability characteristics when compared to a commercially available batch of A. niger glucose oxidase. The gene encoding the new glucose oxidase was isolated and DNA sequence analysis of the coding region showed 80% identity to the sequence of a glucose oxidase gene previously published. However, the similarity of the non-coding sequences up- and downstream of the open reading frame was much less, showing only 66% and 50% identity respectively. Despite the low degree of similarity between the promotor region of the new gene and the previously published one, the new glucose oxidase was likewise induced by calcium carbonate. In addition, we showed that this induction occurred on the transcriptional level. Observations concerning the effect of gluconolactone and the levels of glucose-6-phosphate isomerase upon calcium carbonate induction suggested that the enhancement of glucose oxidase biosynthesis by calcium carbonate was accompanied by a metabolic shift from glycolysis to the pentose phosphate pathway.

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References

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein, utilizing the principle of protein-dye binding. Anal Biochem 12:248–254

    Google Scholar 

  • Fiedurek J, Rogalski J, Ilczuk Z, Leonowicz A (1986) Screening and mutagenesis of moulds for the improvement of glucose oxidase production. Enzyme Microb Technol 8:734–736

    Google Scholar 

  • Frederick KR, Tung J, Emerick RS, Masiarz FR, Chamberlain SH, Vasavada A, Rosenberg S, Chakraborty S, Schopter LM, Massey V (1990) Glucose oxidase from Aspergillus niger: cloning, gene sequence, secretion from Saccharomyces cerevisiae and kinetic analysis of a yeast derived enzyme. J Biol Chem 265:3793–3802

    Google Scholar 

  • Hatzinikolaou DG, Macris BJ (1995) Factors regulating production of glucose oxidase by Aspergillus niger. Enzyme Microb Technol 17:530–534

    Google Scholar 

  • Hellmuth K, Pluschkell S, Jung J-K, Ruttkowski E, Rinas U (1995) Optimization of glucose oxidase production by Aspergillus niger using genetic- and process-engineering techniques. Appl Microbiol Biotechnol 43:978–984

    Google Scholar 

  • Kalisz HM, Hecht H-J, Schomburg D, Schmid RD (1991) Effects of carbohydrate depletion on the structure, stability and activity of glucose oxidase from Aspergillus niger. Biochim Biophys Acta 1080:138–142

    Google Scholar 

  • Kelley RL, Reddy CA (1986) Purification and characterization of glucose oxidase from ligninolytic cultures of Phanerochaete chrysosporium. J Bacteriol 166:269–274

    Google Scholar 

  • Kim KK, Fravel DR, Papavizas GC (1990) Production, purification, and properties of glucose oxidase from the biocontrol fungus Talaromyces flavus. Can J Microbiol 36:199–205

    Google Scholar 

  • Kriechbaum M, Heilmann HJ, Wientjes FJ, Hahn M, Jany KD, Gassen HG, Sharif F, Alaeddinoglu G (1989) Cloning and DNA sequence analysis of the glucose oxidase gene from Aspergillus niger NRRL-3. FEBS Lett 255:63–66

    Google Scholar 

  • Kundu PN, Das A (1985) A note on crossing experiments with Aspergillus niger for the production of calcium gluconate. J Appl Bacteriol 59:1–5

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Markwell J, Frakes LG, Brott EC, Osterman J, Wagner FW (1989) Aspergillus niger mutants with increased glucose oxidase production. Appl Microbiol Biotechnol 30:166–169

    Google Scholar 

  • Muller H-M (1985) Utilization of gluconate by Aspergillus niger. I. Enzymes of phosphorylating and nonphosphorylating pathways. Zentralbl Microbiol 140:475–484

    Google Scholar 

  • Muller H-M (1986) Utilization of gluconate by Aspergillus niger. II. Enzymes of degradation pathways and main end products. Zentralbl Microbiol 140:475–484

    Google Scholar 

  • Pazur JH, Kleppe K (1964) The oxidation of glucose and related compounds by glucose oxidase from Aspergillus niger. Biochemistry 3:578–583

    Google Scholar 

  • Petruccioli M, Federici F (1993) Glucose oxidase production by Penicillium variabile P16: effect of composition. J Appl Bacteriol 75:369–372

    Google Scholar 

  • Petruccioli M, Fenice M, Piccioni P (1993) Distribution and typology of glucose oxidase activity in the genus Penicillium. Lett Appl Microbiol 17:285–288

    Google Scholar 

  • Rogalski J, Fiedurek J, Szczordrak J, Kapusta K, Leonowicz A (1988) Optimization of glucose oxidase synthesis in submerged cultures of Aspergillus niger G-13 mutant. Enzyme Microb Technol 10:508–511

    Google Scholar 

  • Rosenberg S (1993) Polynucleotide sequence for production of glucose oxidase in recombinant systems. U. S. patent 5,094,951

  • Sambrook J, Fritsch EF, Maniatis T (Eds.) (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:54–63

    Google Scholar 

  • Sharif FA, Alaeddinoglu NG (1992) Expression and overproduction of glucose oxidase in Aspergillus niger. Appl Microbiol Biotechnol 38:115–116

    Google Scholar 

  • Simon LM, Kotorman M, Szajani B, Boross L (1986) Preparation and characterization of immobilized glucose-phosphate isomerase. Enzyme Microb Technol 8:222–226

    Google Scholar 

  • Takegawa K, Fujiwara K, Iwahara S, Yamamoto K, Tochikura T (1989) Effect of deglycosylation of N-linked sugar chains on glucose oxidase from Aspergillus niger. Biochem Cell Biol 67:460–463

    Google Scholar 

  • Triglia T, Peterson MG, Kemp DJ (1988) A procedure for in vitro amplification of DNA segments that lie outside the boundaries of known sequences. Nucleic Acids Res 16:81–86

    Google Scholar 

  • Tsuge H, Natsuaki O, Ohashi K (1975) Purification, properties and molecular features of glucose oxidase from Aspergillus niger. J Biochem 78:835–843

    Google Scholar 

  • Whittington H, Kerry-Wiliams S, Bidgood K, Dodsworth N, Peberdy J, Dobson M, Hinchliffe E, Ballance DJ (1990) Expression of the Aspergillus niger glucose oxidase gene in A. niger, A. nidulans and Saccharomyces cerevisiae. Curr Genet 18:531–536

    Google Scholar 

  • Witteveen CFB, Vondervoort P van de, Swart K, Visser J (1990) Glucose oxidase overproducing and negative mutants of Aspergillus niger. Appl Microbiol Biotechnol 33:683–686

    Google Scholar 

  • Witteveen CFB, Vondervoort PJI van de, Broeck HC van den, Engelenburg FAC van, Graaff LH de, Hillebrand MHBC, Schaap PJ, Visser J (1993) Induction of glucose oxidase, catalase, and lactonase in Aspergillus niger. Curr Genet 24:408–416

    Google Scholar 

  • Zetelaki KZ (1970) The role of aeration and agitation in the production of glucose oxidase in submerged culture. II. Biotechnol Bioeng 12:379–397

    Google Scholar 

  • Zetelaki K, Vas K (1968) The role of aeration and agitation in the production of glucose oxidase in submerged culture. Biotechnol Bioeng 10:45–59

    Google Scholar 

Download references

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Author notes

  1. A. Tingey

    Present address: School of Animal and Microbial Sciences, The University of Reading, PO Box 228, RG6 6AJ, Whiteknights, Reading, UK

  2. P. Goodenough

    Present address: School of Plant Sciences, The University of Reading, PO Box 221, RG6 6AS, Whiteknights, Reading, UK

Authors and Affiliations

  1. Biosystems Technology Laboratory, Department of Chemical Engineering, National Technical University of Athens, Zografou Campus, 15780, Athens, Greece

    D. G. Hatzinikolaou, B. J. Macris & D. Kekos

  2. Biotechnological Institute, Anker Engelunds Vej 1, Building 227, DK-2800, Lyngby, Denmark

    O. C. Hansen & P. Stougaard

  3. Reading Laboratory, AFRC Institute of Food Research, Early Gate, Whiteknights Road, RG6 2EF, Reading, UK

    A. Tingey & P. Goodenough

Authors
  1. D. G. Hatzinikolaou
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  2. O. C. Hansen
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  3. B. J. Macris
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  4. A. Tingey
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  5. D. Kekos
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  6. P. Goodenough
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  7. P. Stougaard
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Hatzinikolaou, D.G., Hansen, O.C., Macris, B.J. et al. A new glucose oxidase from Aspergillus niger: characterization and regulation studies of enzyme and gene. Appl Microbiol Biotechnol 46, 371–381 (1996). https://doi.org/10.1007/BF00166232

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  • DOI: https://doi.org/10.1007/BF00166232

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