A Model of Citric Acid Production in the Mold <span class='italic'>Aspergillus niger</span>

Néstor V. Torres; Eberhard O. Voit

doi:10.1017/CBO9780511546334.004

3 - A Model of Citric Acid Production in the Mold Aspergillus niger

Published online by Cambridge University Press: 28 July 2009

Néstor V. Torres and

Eberhard O. Voit

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Néstor V. Torres: Affiliation:
Universidad de la Laguna, Tenerife
Eberhard O. Voit: Affiliation:
Medical University of South Carolina

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INTRODUCTION

This chapter addresses the question of how mathematical modeling can use detailed and localized biochemical information to deepen our understanding of biological functioning at the level of integrated metabolic pathways. It describes the cyclical process of mathematical modeling with the example of citric acid metabolism in Aspergillus niger. This system has some nice features that facilitate mathematical analysis and provide a good example for demonstrating the various steps of model design, analysis, and refinement. Although citric acid metabolism is quite complex and involves several biochemical pathways in two different cellular compartments, the system is relatively closed. Moreover, under conditions where A. niger synthesizes citric acid, this synthesis is essentially the only metabolic pathway of quantitative importance, the substrate is almost entirely glucose and/or fructose, and citric acid is the sole final product. The system is rich in external and internal controls, and the governing biochemical mechanisms are well documented with a comprehensive body of kinetic information. The intricate regulatory structure is a source of nonlinear responses that make nonmathematical approaches unsuitable.

Citric acid production in A. niger was first approached with mathematical modeling techniques by Meyrath (1967). He used a macroscopic approach based on mass and energy balances and concluded that up to 85% of the consumed hexoses could be converted into citric acid. Verhoff and Spradlin (1976) also presented a mass balance model with the aim of unraveling the metabolic pathways involved in citric acid biosynthesis.

Type: Chapter
Information: Pathway Analysis and Optimization in Metabolic Engineering , pp. 75 - 133

DOI: https://doi.org/10.1017/CBO9780511546334.004 [Opens in a new window]

Publisher: Cambridge University Press

Print publication year: 2002

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References

Aiba, S., and Matsuoka, M.: Identification of metabolic model: Citrate production from glucose by Candida lipolytica. Biotechnol. Bioeng. 21, 1373–86, 1979CrossRef Google Scholar

Alvarez-Vasquez, F.: Modelización, análisis y optimización del metabolismo del hongo Aspergillus niger en condiciones de producción de ácido cítrico. PhD dissertation, University of La Laguna, Spain, 2000

Alvarez-Vasquez, F., González-Alcón, C., and Torres, N. V.: Metabolism of citric acid production by Aspergillus niger: Model definition, steady state analysis and constrained optimization of the citric acid production rate. Biotechnol. Bioeng. 70(1), 82–108, 20003.0.CO;2-V>CrossRef Google Scholar PubMed

Amaranisingham, C. F., and Davis, B. D.: Regulation of alpha-ketoglutarate dehydrogenase formation in Escherichia coli. J. Biol. Chem. 240, 3664–8, 1965Google Scholar

Arisan-Atac, I., Wolschek, M., and Kubicek, C. P.: Trehalose-6-phosphate synthase A affects citrate accumulation by Aspergillus niger under conditions of high glycolytic flux. FEMS Microbiol. Lett. 140, 77–83, 1996CrossRef Google Scholar PubMed

Arts, E., Kubicek, C. P., and Röhr, M.: Regulation of phosphofructokinase from Aspergillus niger: Effect of fructose 2,6 bisphosphate on the action of citrate, ammonium ions and AMP. J. Gen. Microbiol. 133, 1195–9, 1987Google Scholar

Blázquez, M. A., Lagunas, R., Gancedo, C., and Gancedo, J. M.: Trehalose-6-phosphate, a new regulator of yeast glycolysis that inhibits hexokinases. FEBS Lett. 329, 51–4, 1993CrossRef Google Scholar PubMed

Bloom, S., and Johnson, M.: The pyruvate carboxylase of Aspergillus niger. J. Biol. Chem. 237, 2718–20, 1962Google Scholar PubMed

Boy-Marcotte, E., Lagniel, G., Perrot, M., Bussereau, F., Boudsocq, A., Jacquet, M., and Labarre, J.: The heat shock response in yeast: Differential regulations and contributions of the Msn2p/Msn4p and Hsf1p regulons. Mol. Microbiol. 33(2), 274–83, 1999CrossRef Google Scholar PubMed

Briquet, M.: Transport of pyruvate and lactate in yeast mitochondria. Biochim. Biophys. Acta 459, 290–9, 1977CrossRef Google Scholar PubMed

Cleland, W. W., and Johnson, M. J.: Tracer experiments on the mechanism of citric acid formation by Aspergillus niger. J. Biol. Chem. 208, 679–92, 1954Google Scholar PubMed

Crow, V. L., and Wittenberger, C. L.: Separation and properties of NAD+- and NADP+-dependent glyceraldehyde-3-phosphate dehydrogenases from Streptococcus mutans. J. Biol. Chem. 254(4), 1134–42, 1979Google Scholar PubMed

Evans, C. T., Scragg, A. H., and Ratledge, C.: Regulation of citrate efflux from mitochondria of oleaginous and non-oleaginous yeast by adenine nucleotides. Eur. J. Biochem. 132, 609–15, 1981CrossRef Google Scholar

Evans, C. T., Scragg, A. H., and Ratledge, C.: A comparative study of citrate efflux from mitochondria of oleaginous and non-oleaginous yeast. Eur. J. Biochem. 130, 195–204, 1983CrossRef Google Scholar

Feir, H. A., and Suzuki, I.: Pyruvate carboxilase of Aspergillus niger: Kinetic study of a biotin-containing carboxilase. Can. J. Biochem. 47, 697–710, 1969CrossRef Google Scholar

Fell, D. A.: Metabolic control analysis – a survey of its theoretical and experimental development. Biochem. J. 286, 313–30, 1992CrossRef Google Scholar PubMed

Ferreira, A.: PLAS©: http://correio.cc.fc.ul.pt/~aenf/plas.html, 2000

Führer, L., Kubicek, C. P., and Röhr, M.: Pyridine nucleotide levels and ratios in Aspergillus niger. Can. J. Biochem. 26, 405–8, 1980Google Scholar PubMed

Groen, A. K., R. J. A. Wanders, H. V. Westerhoff, R. van der Meer, and J. M. Tager: Control of metabolic fluxes. In: H. Sies (Ed.), Metabolic Compartmentation (pp. 9–37). Academic Press, New York, 1982

Guebel, D., and Torres, N. V.: Optimization of the citric acid production by A. niger through a metabolic flux balance model. Elect. J. Biotechnol., in press

Habison, A., Kubicek, C. P., and Röhr, M.: Phosphofructokinase as a regulatory enzyme in citric acid producing Aspergillus niger. FEMS Microbiol. Lett. 5, 39–42, 1979CrossRef Google Scholar

Habison, A., Kubicek, C. P., and Röhr, M.: Partial purification and regulatory properties of phosphofructokinase from Aspergillus niger. Biochem. J. 209, 669–76, 1983CrossRef Google Scholar PubMed

Halestrap, A. P., Scott, R. D., and Thomas, A. P.: Mitochondrial pyruvate transport and its hormonal regulation. Int. J. Biochem. 11, 97–105, 1980CrossRef Google Scholar PubMed

Henry, M-F., and Nyns, E.-J.: Cyanide-insensitive respiration. An alternative mitochondrial pathway. Sub-Cell. Biochem. 4, 1–65, 1975Google Scholar PubMed

Henson, C. P., and Cleland, W. W.: Kinetic studies of glutamic-oxalacetate transaminase isoenzymes. Biochemistry 3, 338–48, 1964CrossRef Google Scholar

Jaklitsch, W., Kubicek, C. P., and Scrutton, M.: Intracellular location of enzymes involved in citrate production by Aspergillus niger. Can. J. Microbiol. 37, 823–7, 1991CrossRef Google Scholar PubMed

Kirimura, K., Hirowatari, Y., and Usami, S.: Alterations of respiratory systems in Aspergillus niger under the condition of citric acid fermentation. Agric. Biol. Chem. 51, 1299–303, 1987Google Scholar

Kirimura, K., Yoda, M., Shimizu, H., Sugano, S., Mizuno, M., Kino, K., and Usami, S.: Contribution of cyanide-insensitive respiratory pathway, catalyzed by the alternative oxidase to citric acid production in Aspergillus niger. Biosci. Biotechnol. Biochem. 64(10), 2034–9, 2000CrossRef Google Scholar PubMed

Kubicek, C. P.: Aspects of control of metabolic fluxes in microorganisms. Dechema-Monogr., VCH Verlagsgesellschaft 105, 81–95, 1987Google Scholar

Kubicek, C. P.: Organic acids. In: C. Ratledge and B. Kristiansen (Eds.), Basic Biotechnology (Chapter 14). Cambridge University Press, Cambridge, U.K., 2001

Kubicek, C. P., and Röhr, M.: Influence of manganese on enzyme synthesis and citric acid accumulation in Aspergillus niger. Eur. J. Appl. Microbiol. 4, 167–173, 1977CrossRef Google Scholar

Kubicek, C. P., and Röhr, M.: The role of the tricarboxylic acid cycle in citric acid accumulation by Aspergillus niger. Eur. J. Appl. Microbiol. Biotechnol. 5, 263–71, 1978CrossRef Google Scholar

Kubicek, C. P., and Röhr, M.: Citric acid fermentation. CRC Crit. Rev. Biotechnol. 3(4), 331–73, 1986CrossRef Google Scholar

Kubicek, C. P., Hampel, W., and Röhr, M.: Manganese deficiency leads to elevated amino acid pools in citric acid accumulating Aspergillus niger. Arch. Microbiol. 123, 73–9, 1979CrossRef Google Scholar PubMed

Kubicek, C. P., Zehentgruber, O., El-Kalak, H., and Röhr, M.: Regulation of citric acid production by oxygen: Effects of dissolved oxygen tension on adenylate levels and respiration in Aspergillus niger. Eur. J. Appl. Microbiol. Biotechnol. 9, 101–16, 1980CrossRef Google Scholar

Kubicek-Pranz, E. M., Mozelt, M., Roehr, M., and Kubicek, C. P.: Changes in the concentration of fructose 2, 6-bisphosphate in Aspergillus niger during stimulation of acidogenesis by elevated sucrose concentration. Biochim. Biophys. Acta 1033, 250–5, 1990CrossRef Google Scholar PubMed

Legisa, M., and Mattey, M.: Glycerol synthesis by Aspergillus niger under citric acid accumulating conditions. Enzyme Microbiol. Technol. 8, 607–9, 1988CrossRef Google Scholar

Ma, H., Kubicek, C. P., and Röhr, M.: Malate dehydrogenase isoenzymes in Aspergillus niger. FEMS Microbiol. Lett. 12, 147–51, 1981CrossRef Google Scholar

Mattey, M.: Citrate regulation of citric acid production by Aspergillus niger. FEMS Microbiol. Lett. 2, 71–4, 1977CrossRef Google Scholar

Mattey, M.: The production of organic acids. Crit. Rev. Biotechnol. 12, 87–132, 1992CrossRef Google Scholar PubMed

Meixner-Monori, B., Kubicek, C. P., and Röhr, M.: Pyruvate kinase from Aspergillus niger: A regulatory enzyme in glycolysis?Can. J. Microbiol. 30, 16–22, 1983CrossRef Google Scholar

Meixner-Monori, B., Kubicek, C. P., Habison, A., Kubicek-Pranz, E. M., and Röhr, M.: Presence and regulation of the α-ketoglutarate dehydrogenase multienzyme complex in the filamentous fungus Aspergillus niger. J. Bacteriol. 161, 265–71, 1985Google Scholar PubMed

Meixner-Monori, B., Kubicek, C. P., Harrer, W., Schreferl, G., and Röhr, M.: NADP-specific isocitrate dehydrogenase from the citric acid-accumulating fungus Aspergillus niger. Biochem. J. 236, 549–57, 1986CrossRef Google Scholar PubMed

Meyrath, J.: Citric acid production. Proc. Biochem. 2, 25–7, 1967Google Scholar

Mischak, H., Kubicek, C. P., and Röhr, M.: Citrate inhibition of glucose uptake in Aspergillus niger. Biotechnol. Lett. 6, 425–30, 1984CrossRef Google Scholar

Netik, A., Torres, N. V., Riol, J. M., and Kubicek, C. P.: Uptake and export of citric acid by Aspergillus niger is reciprocally regulated by manganese ions. Biochim. Biophys. Acta 1326, 287–94, 1997CrossRef Google Scholar PubMed

Newsholme, E. A., and C. Start: Regulation in Metabolism. John Wiley & Sons, London, 1973

Ni, T.-C., and Savageau, M. A.: Application of biochemical systems theory to metabolism in human red blood cells. Signal propagation and accuracy of representation. J. Biol. Chem. 271(14), 7927–41, 1996aCrossRef Google Scholar

Ni, T.-C., and Savageau, M. A.: Model assessment and refinement using strategies from biochemical systems theory: Application to metabolism in human red blood cells. J. Theor. Biol. 179, 329–68, 1996bCrossRef Google Scholar

Osmani, S. A., and Scrutton, M. C.: The subcellular localization of pyruvate carboxylase and of some other enzymes in Aspergillus nidulans. Eur. J. Biochem. 133, 551–60, 1983CrossRef Google Scholar

Panneman, H., Ruijter, G. J. G., Broeck, H. C., Driever, E. T., and Visser, J.: Cloning and biochemical characterization of an Aspergillus niger glucokinase. Evidence for the presence of separate glucokinase and hexokinase enzymes. Eur. J. Biochem. 240, 518–25, 1996CrossRef Google Scholar

Panneman, H., Ruijter, G. J. G., Broeck, H. C., and Visser, J.: Cloning and biochemical characterization of Aspergillus niger hexokinase. The enzyme is strongly inhibited by physiological concentrations of trehalose 6-phosphate. Eur. J. Biochem. 258, 223–32, 1998CrossRef Google Scholar

Pedersen, H., Carlsen, M., and Nielsen, J.: Identification of enzymes and quantification of metabolic fluxes in the wild type and in a recombinant Aspergillus oryzae strain. Appl. Environ. Microbiol. 65(1), 11–19, 1999Google Scholar

Perkins, M., Haslam, J. M., and Linnane, A. W.: Biogenesis of mitochondria. The effects of physiological and genetic manipulation of Saccharomyces cerevisiae on the mitochondrial transport sysytem for tricarboxylate-cycle anions. Biochem. J. 134, 923–934, 1973CrossRef Google Scholar

Prömper, C., Schneider, R., and Weiss, H.: The role of the proton-pumping and alternative respiratory chain NADH: Ubiquinone oxidoreductases in overflow catabolism of Aspergillus niger. Eur. J. Biochem. 216, 223–30, 1993CrossRef Google Scholar PubMed

Reich, J. G., and E. E. Sel'kov: Energy Metabolism of the Cell. Academic Press, London, 1981

Röhr, M.: A century of citric acid fermentation and research. Food Technol. Biotechnol. 36(3), 163–71, 1998Google Scholar

Röhr, M., and Kubicek, C. P.: Regulatory aspects of citric acid fermentation by Aspergillus niger. Proc. Biochem. 16, 34–7, 1981Google Scholar

Röhr, M., Kubicek, C. P., Zehentgruber, O., and Orthofer, R.: Accumulation and partial re-consumption of polyols during citric acid fermentation by Aspergillus niger. Appl. Microbiol. Biotechnol. 27, 235–9, 1987CrossRef Google Scholar

Sakai, K., Hasumi, K., and Endo, A.: Two glyceraldehyde-3-phosphate dehydrogenase isozymes from the koningic acid (heptelidic acid) producer Trichoderma koningii. Eur. J. Biochem. 193(1), 195–202, 1990CrossRef Google Scholar PubMed

Salvador, A.: Synergism analysis of biochemical systems. I. Conceptual framework. Math. Biosci. 163(2), 105–29, 2000aCrossRef Google Scholar

Salvador, A.: Synergism analysis of biochemical systems. II. Tensor formulation and treatment of stoichiometric constraints. Math. Biosci. 163(2), 131–58, 2000bCrossRef Google Scholar

Savageau, M. A.: Biochemical systems analysis. II. The steady-state solutions for an n-pool system using a power-law approximation. J. Theor. Biol. 25, 370–9, 1969CrossRef Google Scholar PubMed

Savageau, M. A.: Optimal design of feedback control by inhibition: Dynamic considerations. J. Mol. Evol. 5, 199–222, 1975CrossRef Google Scholar PubMed

Savageau, M. A.: Biochemical System Analysis: A study of Function and Design in Molecular Biology. Addison-Wesley, Reading, MA, 1976

Schmidt, M., Wallrath, J., Dörner, A., and Weiss, H.: Disturbed assembly of the respiratory chain NADH: Ubiquinone reductase (complex I) in citric-acid-accumulating Aspergillus niger strain B60. Appl. Microbiol. Biotechnol. 36, 667–72, 1992CrossRef Google Scholar

Schreferl-Kunar, G., Grotz, M., Röhr, M., and Kubicek, C. P.: Increased citric acid production by mutants of Aspergillus niger with increased glycolytic capacity. FEMS Microbiol. Lett. 59, 297–300, 1989CrossRef Google Scholar

Shiraishi, F., and Savageau, M. A.: The tricarboxylic acid cycle in Dictyostelium discoideum. I. Formulation of alternative kinetic representations. J. Biol. Chem. 267(32), 22912–18, 1992aGoogle Scholar

Shiraishi, F., and Savageau, M. A.: The tricarboxylic acid cycle in Dictyostelium discoideum. II. Evaluation of model consistency and robustness. J. Biol. Chem. 267(32), 22919–25, 1992bGoogle Scholar

Shiraishi, F., and Savageau, M. A.: The tricarboxylic acid cycle in Dictyostelium discoideum. III. Analysis of steady state and dynamic behavior. J. Biol. Chem. 267(32), 22926–33, 1992cGoogle Scholar

Sorribas, A., and Savageau, M. A.: Strategies for representing metabolic pathways within biochemical systems theory: Reversible pathways. Math. Biosci. 94, 239–69, 1989CrossRef Google Scholar PubMed

Steinbock, F. A.: Doctoral dissertation, Technical University of Vienna, Vienna, Austria, 1993

Steinbock, F. A., Held, I., Chojun, S., Harsen, H., Röhr, M., Kubicek-Prantz, E. M., and Kubicek, C. P.: Regulatory aspects of carbohydrate metabolism in relation to citric acid accumulation by Aspergillus niger. Acta Biotechnol. 116, 571–81, 1991CrossRef Google Scholar

Supply, P., Wach, A., and Goffeau, A.: Enzymatic properties of the PMA2 plasma membrane-bound H+-ATPase of Saccharomyces cerevisiae. J. Biol. Chem. 268(25), 19753–9, 1993Google Scholar PubMed

Torres, N. V.: Modeling approach to control of carbohydrate metabolism during citric acid accumulation by Aspergillus niger. I. Model definition and stability of the steady state. Biotechnol. Bioeng. 44, 104–11, 1994aCrossRef Google Scholar

Torres, N. V.: Modeling approach to control of carbohydrate metabolism during citric acid accumulation by Aspergillus niger. II. Sensitivity analysis. Biotechnol. Bioeng. 44, 112–18, 1994bCrossRef Google Scholar

Torres, N. V., Riol-Cimas, J. M., Wolschek, M., and Kubicek, C. P.: Glucose transport by Aspergillus niger: The low-affinity carrier is only formed during growth on high glucose concentrations. Appl. Microbiol. Biotechnol. 44, 790–4, 1996Google Scholar

Verhoff, F. H., and Spradlin, J. E.: Mass and energy balance analysis of metabolic pathways applied to citric acid production by Aspergillus niger. Biotechnol. Bioeng. 18(3), 425–32, 1976CrossRef Google Scholar PubMed

Voit, E. O.: Computational Analysis of Biochemical Systems. A Practical Guide for Biochemists and Molecular Biologists (ⅻ + 533 pp.). Cambridge University Press, Cambridge, U.K., 2000

Voit, E. O., and Ferreira, A. E. N.: Buffering in models of integrated biochemical systems. J. Theor. Biol. 191, 429–38, 1998CrossRef Google Scholar

Wallrath, J., Schmidt, J., and Weiss, H.: Concomitant loss of respiratory chain NADH: ubiquinone reductase (complex I) and citric acid accumulation in Aspergillus niger. Appl. Microbiol. Biotechnol. 36, 76–81, 1991CrossRef Google Scholar

Wehmer, C.: Beiträge zur Kenntnis einheimischer Pilze. I. Zwei neue Schimmelpilze als Erreger einer Citronensäure-Gärung. Halm, Hannover/Leipzig, 1903

Wolschek, M. F., and C. P. Kubicek: Biochemistry of citric acid accumulation in Aspergillus niger. In: B. Kristiansen, M. Mattey, and J. Linden (Eds.), Citric Acid Biotechnology (pp. 11–32). Taylor and Francis, London, U.K., 1998

Woronick, C., and Johnson, M. J.: Carbon dioxide fixation by cell-free extracts of Aspergillus niger. J. Biol. Chem. 235, 9–15, 1960Google Scholar PubMed

Xu, D.-B., Madrid, C. P., Röhr, M., and Kubicek, C. P.: The influence of type and concentration of the carbon source on production of citric acid by Aspergillus niger. Appl. Microbiol. Biotechnol. 30, 553–8, 1989Google Scholar

Zahorsky, B.: U. S. Patent 1,065,358, 1913

Zehentgruber, O., Kubicek, C. P., and Röhr, M.: Alternative respiration of Aspergillus niger. FEMS Microbiol. Lett. 8, 71–4, 1980CrossRef Google Scholar

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