Process Biotechnology
EJB Electronic Journal of Biotechnology ISSN: 0717-3458 Vol.4 No. 1, Issue of April 15, 2001
© 2001 by Universidad Católica de Valparaíso -- Chile Received October 27, 2000 / Accepted January 6, 2001
RESEARCH ARTICLE

Optimization of the citric acid production by Aspergillus niger through a metabolic flux balance model

Daniel V. Guebel
Counseling Biotechnological Services
Av. San Martin 4927 Dpto. A. (C1417DSJ)
Buenos Aires. Argentina
Tel: 54-11-45039355
E-mail: dguebel@radar.com.ar

Néstor V. Torres Darias*
Grupo de Tecnología Bioquímica y Control Metabólico
Dpto. de Bioquímica y Biología Molecular
Facultad de Biología, Universidad de La Laguna
38206 La Laguna, Tenerife, Islas Canarias, España
Tel and fax: +34-922-318334
E-mail: ntorres@ull.es

*Corresponding author

Financial support: The work of one of us (N.V.T.D) was supported by a research grant from the Comisión Interministerial de Ciencia y Tecnología, contract nº BIO99-0492-C02-02 and also by a research from the Gobierno de Canarias, contract nº PI2000-071.

Keywords: bioenergetics, metabolic engineering, metabolic pathway, stoichiometry.

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Idiophase, the citric acid producing stage of Aspergillus niger was mathematically modeled to identify required genetic manipulations to optimize citric acid production rate. For this reason, a consistent picture of cell functioning had to be achieved. The transient idiophase nature was established by stoichiometric analysis. The main intracellular fluxes were computed by application of material and physiological constraints (ATP, reduction equivalents, proton motive force) at culture time 120 hours. The HMP pathway accounts for 16% of the glucose input (carbon basis), the Krebs cycle for 13% and the citric acid synthesis for the remaining 71%. This profile implies an operative glycerol-P shuttle. It recycles 93% of the cytosolic glycerol-P to cytosolic DHAP thus coupling the transformation of cytosolic NADH to mitochondrial FADH. A cellular maintenance energy of 3.7 mmol ATP/g·h was determined. It would be spent in fueling cytoplasmatic (1.4 mmol H+/g·h) and mitochondrial (1.8 mmol H+/g·h) H+-ATPase pumps with efficiencies of 0.65 and 1.2 mmol H+/mmol ATP respectively. The role and extent of the alternative respiration system activity and polyol excretion is accounted by the model as well. In addition, the significance of GABA shunt and futile NH4+/NH3 cycle were rejected. According to the developed model, the specific citric productivity would be increased in 45% by an unique change if glucose influx were duplicated. Differences with predictions from other model that required many manipulations are also discussed.

 
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