Skip to main content
Springer Nature Link
Log in

The role of glucosamine-6-phosphate deaminase at the early stages of Aspergillus niger growth in a high-citric-acid-yielding medium

  • Biotechnologically Relevant Enzymes and Proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Glucosamine-6-phosphate (GlcN6P) deaminase seems to be the main enzyme in Aspergillus niger cells responsible for rapid glucosamine accumulation during the early stages of growth in a high-citric-acid-yielding medium. By determining basic kinetic parameters on the isolated enzyme, a high affinity toward fructose-6-phosphate (Fru6P) was measured, while in the reverse direction the K m value for glucosamine-6-phosphate was lower than deaminases from other organisms measured so far. The enzyme characteristics of GlcN6P deaminase suggest it must compete with 6-phosphofructo-1-kinase (PFK1) for the common substrate—Fru6P in A. niger cells. Glucosamine accumulation seems therefore to remove an intermediate from the glycolytic flux, a situation which is reflected in slower citric acid accumulation and a specific growth rate after the germination of spores. When ammonium ions are depleted from the medium, one of the substrates for GlcN6P deaminase becomes limiting and Fru6P can be catabolised by PFK1 which enhances glycolytic flux. Other enzymatic features of GlcN6P deaminase such as pH optima for both aminating and deaminating reactions might play a significant role in rapid glucosamine accumulation during the early phase of fermentation and a slow consumption of aminosugar during the citric-acid-producing phase.

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

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Explore related subjects

Discover the latest articles, books and news in related subjects, suggested using machine learning.

References

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein–dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Calcagno M, Campos PJ, Mulliert G, Suastegui J (1984) Purification, molecular and kinetic properties of glucosamine-6-phosphate isomerase (deaminase) from Escherichia coli. Biochim Biophys Acta 787:165–173

    Article  CAS  Google Scholar 

  • Elson LA, Morgan WT (1933) A colorimetric method for the determination of glucosamine and chondrosamine. Biochem J 27:1824–1828

    CAS  PubMed  PubMed Central  Google Scholar 

  • Enghofer E, Kress H, Linzen B (1978) Glucosamine metabolism in Drosophila salivary glands. Separation of metabolites and some characteristics of three enzymes involved. Biochim Biophys Acta 544:245–251

    Article  CAS  Google Scholar 

  • Ghosh S, Blumenthal HJ, Davidson E, Roseman S (1960) Glucosamine metabolism, Enzymatic synthesis of glucosamine-6-phosphate. J Biol Chem 235:1265–1273

    CAS  PubMed  Google Scholar 

  • Gradišnik-Grapulin M, Legiša M (1997) A spontaneous change in the intracellular cyclic AMP level in Aspergillus niger is influenced by the sucrose concentration in the medium and by light. Appl Environ Microbiol 63:2844–2849

    Article  Google Scholar 

  • Hondmann DHA, Visser J (1994) Carbon metabolism. In: Martinelli SD, Kinghorn JR (eds) Aspergillus: 50 years on, progress in industrial microbiology. vol. 29. Elsevier, Amsterdam, pp 59–139

    Google Scholar 

  • Jernejc K, Legiša M (2004) A drop of intracellular pH stimulates citric acid accumulation by some strains of Aspergillus niger. J Biotechnol 112:289–297

    Article  CAS  Google Scholar 

  • Kubicek CP, Hampel W, R, hr M (1979) Manganese deficiency leads to elevated amino acid pools in citric acid accumulating Aspergillus niger. Arch Microbiol 123:73–79

    Article  CAS  Google Scholar 

  • Kumar S, Punekar NS, SatyaNarayan V, Venkatesh KV (2000) Metabolic fate of glutamate and evaluation of flux through the 4-aminobutyrate (GABA) shunt in Aspergillus niger. Biotechnol Bioeng 67:575–584

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Lagorce A, Berre-Anton V, Aguilar-Uscanga B, Martin-Yken H, Dagkessamanskaia A, Francois J (2002) Involvement of GFA1, which encodes glutamine-fructose-6-phosphate amidotransferase, in the activation of the chitin synthesis pathway in response to cell-wall defects in Saccharomyces cerevisiae. Eur J Biochem 269:1697–1707

    Article  CAS  Google Scholar 

  • Lara-Lemus R, Calcagno ML (1998) Glucosamine-6-phosphate deaminase from beef kidney is an allosteric system of the V-type. Biochim Biophys Acta 1388:1–9

    Article  CAS  Google Scholar 

  • Legiša M, Mattey M (1986) Glycerol synthesis by Aspergillus niger under citric acid accumulating conditions. Enzyme Microb Technol 8:607–609

    Article  Google Scholar 

  • Legiša M, Kidrič J (1989) Initiation of citric acid accumulation in the early stages of Aspergillus niger growth. Appl Microbiol Biotechnol 31:453–457

    Article  Google Scholar 

  • Legiša M, Benčina M (1994) Evidence for the activation of 6-phosphofructo-1-kinase by cAMP-dependent protein kinase in Aspergillus niger. FEMS Microbiol Lett 118:327–333

    Article  Google Scholar 

  • Legiša M, Mattey M (2007) Changes in primary metabolism leading to citric acid overflow in Aspergillus niger. Biotechnol Lett 29:181–190

    Article  Google Scholar 

  • Levvy GA, McAllan A (1959) The N-acetylation and estimation of hexosamines. Biochem J 73:127–132

    PubMed Central  Google Scholar 

  • Mesojednik S, Legiša M (2005) Posttranslational modification of 6-phosphofructo-1-kinase in Aspergillus niger. Appl Environ Microbiol 71:1425–1432

    Article  CAS  Google Scholar 

  • Mlakar T, Legiša M (2006) Citrate inhibition-resistant form of 6-phosphofructo-1-kinase from Aspergillus niger. Appl Environ Microbiol 72:4515–4521

    Article  CAS  Google Scholar 

  • Papagianni M, Wayman F, Mattey M (2005) Fate and role of ammonium ions during fermentation of citric acid by Aspergillus niger. Appl Environ Microbiol 71:7178–7186

    Article  CAS  Google Scholar 

  • Pattabiraman TN, Bachhawat BK (1961) Purification of glucosamine 6-phosphate deaminase from human brain. Biochim Biophys Acta 54:273–283

    Article  CAS  Google Scholar 

  • Peksel A, Torres NV, Liu J, Juneau G, Kubicek CP (2002) 13C-NMR analysis of glucose metabolism during citric acid production by Aspergillus niger. Appl Microbiol Biotechnol 58:157–163

    Article  CAS  Google Scholar 

  • Pel HJ, de Winde JH, Archer DB, Dyer PS, Hofmann G, Schaap PJ, Turner G, de Vries RP, Albang R, Albermann K, Andersen MR, Bendtsen JD, Benen JA, van den Berg M, Breestraat S, Caddick MX, Contreras R, Cornell M, Coutinho PM, Danchin EG, Debets AJ, Dekker P, van Dijck PW, van Dijk A, Dijkhuizen L, Driessen AJ, d'Enfert C, Geysens S, Goosen C, Groot GS, de Groot PW, Guillemette T, Henrissat B, Herweijer M, van den Hombergh JP, van den Hondel CA, van der Heijden RT, van der Kaaij RM, Klis FM, Kools HJ, Kubicek CP, van Kuyk PA, Lauber J, Lu X, van der Maarel MJ, Meulenberg R, Menke H, Mortimer MA, Nielsen J, Oliver SG, Olsthoorn M, Pal K, van Peij NN, Ram AF, Rinas U, Roubos JA, Sagt CM, Schmoll M, Sun J, Ussery D, Varga J, Vervecken W, van de Vondervoort PJ, Wedler H, Wosten HA, Zeng AP, van Ooyen AJ, Visser J, Stam H (2007) Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.8. Nat Biotechnol 25:221–231

    Article  Google Scholar 

  • Ram AFJ, Arentshorst M, Damveld RA, vanKuyk PA, Klis FM, van den Hondel CAMJJ (2004) The cell wall stress response in Aspergillus niger involves increased expression of the glutamine: fructose-6-phosphate amidotransferase-encoding gene (gfaA) and increased deposition of chitin in the cell wall. Microbiology 150:3315–3326

    Article  CAS  Google Scholar 

  • Röhr M, Kubicek CP (1981) Regulatory aspects of citric acid fermentation by Aspergillus niger. Process Biochem 16:34–37

    Google Scholar 

  • Ruijter GJG, Panneman H, Visser J (1997) Overexpression of phosphofructokinase and pyruvate kinase in citric acid-producing Aspergillus niger. Biochim Biophys Acta 1334:317–326

    Article  CAS  Google Scholar 

  • Singh P, Ghosh D, Datta A (2001) Attenuation of virulence and changes in morphology in Candida albicans by disruption of the N-acetylglucosamine catabolic pathway. Infect Immun 69:7898–7903

    Article  CAS  Google Scholar 

  • Tanaka T, Takahashi F, Fukui T, Fujiwara S, Atomi H, Imanaka T (2005) Characterization of a novel glucosamine-6-phosphate deaminase from a hyperthermophilic archaeon. J Bacteriol 187:7038–44

    Article  CAS  Google Scholar 

  • Vincent F, Davies GJ, Brannigan JA (2005) Structure and kinetics of a monomeric glucosamine 6-phosphate deaminase: missing link of the NagB superfamily? J Biol Chem 280:19649–19655

    Article  CAS  Google Scholar 

  • Vishniac W, Santer M (1957) The thiobacilli. Bacteriol Rev 21:195–213

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Dr. Michael Mattey for critical reading of the manuscript. We are grateful to the DSM Research B.V., The Netherlands, for providing the A. niger GlcN6P deaminase nucleotide and deduced amino acid sequence prior to publishing the data on A. niger genome.

The work was supported by the Ministry of Higher Education, Science and Technology of the Republic of Slovenia.

Author information

Authors and Affiliations

  1. National Institute of Chemistry, Hajdrihova 19, 1001, Ljubljana, Slovenia

    Tina Šolar, Janja Turšič & Matic Legiša

Authors
  1. Tina Šolar
    View author publications

    Search author on:PubMed Google Scholar

  2. Janja Turšič
    View author publications

    Search author on:PubMed Google Scholar

  3. Matic Legiša
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Matic Legiša.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Šolar, T., Turšič, J. & Legiša, M. The role of glucosamine-6-phosphate deaminase at the early stages of Aspergillus niger growth in a high-citric-acid-yielding medium. Appl Microbiol Biotechnol 78, 613–619 (2008). https://doi.org/10.1007/s00253-007-1339-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-007-1339-4

Keywords