Skip to main content
SpringerLink
Log in

Detection of endogenous β-glucuronidase activity in Aspergillus niger

  • Original Paper
  • Applied Microbial and Cell Physiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

An endogenous β-glucuronidase that hydrolyses the chromogenic substrate 5-bromo-4-chloro-3-indolyl-β-d-glucuronide (X-gluc) in Aspergillus niger is reported. The activity was induced when the fungus was grown in media containing xylan, but was either very low, or absent, when grown on glucose. Endogenous β-glucuronidase was primarily located in newly formed hyphae, and was apparent at pH values between 3 and 6. Hydrolysis of X-gluc was sensitive to the inhibitor d-saccharic acid 1,4-lactone and was irreversibly inactivated by heating. The bacterial uidA β-glucuronidase reporter gene was strongly expressed in the hyphae of transformed A. niger but, in contrast to the endogenous activity, the enzyme was also active at pH 7–8.5. Histochemical localization of uidA expression in A. niger, without interference from the endogenous β-glucuronidase activity, was achieved by staining at this pH.

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

Access this article

Log in via an institution

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

  • Bancroft JD, Stevens A (1994) Buffers. In: Bancraft JD, Stevens (eds) Theory and practice of histological techniques. Churchill Livingstone, London, p 697

    Google Scholar 

  • Buehler HJ, Katzman PA, Doisy EA (1951) Studies on β-glucuronidase from E.coli. Proc Soc Exp Biol Med 76: 672–676

    CAS  Google Scholar 

  • de Graaff LH, van den Broeck HC, van Ooijen AJJ, Visser J (1994) Regulation of the xylanase-encoding xlnA gene of Aspergillus tubigensis. Mol Microbiol 12: 479–490

    Article  Google Scholar 

  • Hänsch R, Koprek T, Mendel RR, Schulze J (1995) An improved protocol for eliminating endogenous β-glucuronidase background in barley. Plant Sci 105: 63–69

    Article  Google Scholar 

  • Hodal L, Bochardt A, Nielsen JE, Mattsson O, Okkels FT (1992) Detection, expression and specific elimination of endogenous β-glucuronidase activity in transgenic and non-transgenic plants. Plant Sci 87: 115–122

    Article  CAS  Google Scholar 

  • Hu C, Chee PP, Chesney RH, Zhou JH, Miller PD, O’Brian WT (1990) Intrinsic GUS-like activities in seed plants. Plant Cell Rep 9: 1–5

    Article  CAS  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405

    Article  CAS  Google Scholar 

  • Levvy GA (1952) The preparation and properties of β-glucuronidase. Biochem J 52: 464–472

    CAS  Google Scholar 

  • Levvy GA (1954) Baicalinase, a plant β-glucuronidase. Biochem J 58: 462–469

    CAS  Google Scholar 

  • Muro T, Kuramoto T, Imoto K, Okada S (1986) Purification and some properties of glycyrrhizinic acid hydrolase from Aspergillus niger GRM3. Agric Biol Chem 50: 687–692

    CAS  Google Scholar 

  • Plegt L, Bino RJ (1989). β-Glucuronidase activity during development of the male gametophyte from transgenic and non-transgenic plants. Mol Gen Genet 216: 321–327

    Article  CAS  Google Scholar 

  • Punt PJ, Greaves PA, Kuyvenhoven A (1991a) A twin reporter vector for simultaneous analysis of expression signals of divergently transcribed, contiguous genes in filamentous fungi. Gene 104: 119–122

    Article  CAS  Google Scholar 

  • Punt PJ, Zegers ND, Busscher M, Pouwels PH, van den Hondel CAMJJ (1991b) Intracellular and extracellular production of proteins under the control of expression signals of the highly expressed Aspergillus nidulans gpdA gene. J Biotechnol 17: 19–34

    Article  CAS  Google Scholar 

  • Roberts IN, Oliver RP, Punt PJ, van den Hondel CA-MJJ (1989) Expression of the Escherichia coli β-glucuronidase gene in industrial and phytopathogenic filamentous fungi. Curr Genet 15: 177–180

    Article  CAS  Google Scholar 

  • de Ruiter-Jacobs YMJT, Broekhuijsen MP, Campbell EI (1989) A gene transfer system based on the homologous pyrG gene and efficient expression of bacterial genes in Aspergillus oryzae. Curr Genet 16: 159–163

    Article  Google Scholar 

  • Verdoes JC, Punt PJ, Stouthamer AH, van den Hondel CAMJJ (1994) The effect of multiple copies of the upstream region on expression of the Aspergillus niger glucoamylase-encoding gene. Gene 145: 179–187

    Article  CAS  Google Scholar 

  • Wozniak CA, Owens LD (1994) Native β-glucuronidase activity in sugarbeet (Beta Vulgaris). Physiol Plant 90: 763–771

    Article  CAS  Google Scholar 

  • Yelton MM, Hamer JE, Timberlake WE (1984). Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad Sci USA 81: 1470–1474

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Danisco Biotechnology, Langebrogade 1, PO Box 17, DK-1001, Copenhagen K, Denmark

    T. E. Gottschalk, J. E. Nielsen & P. Rasmussen

Authors
  1. T. E. Gottschalk
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. E. Nielsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. Rasmussen
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gottschalk, T.E., Nielsen, J.E. & Rasmussen, P. Detection of endogenous β-glucuronidase activity in Aspergillus niger . Appl Microbiol Biotechnol 45, 240–244 (1996). https://doi.org/10.1007/s002530050677

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s002530050677

Keywords

Search

Navigation