Mathematical modelling of a mixed culture cultivation process for the production of polyhydroxybutyrate.

Mixed cultures submitted to acetate "feast" and "famine" cycles are able to store intracellularly high quantities of polyhydroxybutyrate (PHB). It was demonstrated in a previous study that the intracellular PHB content can be increased up to 78.5% (g HB/gVSS) of cell dry weight in a sequencing batch reactor (SBR) with optimised operating conditions. The specific PHB formation rate was also shown to be higher for mixed cultures than for pure cultures. Such high intracellular PHB contents and specific productivity open new perspectives for the industrial production of polyhydroxyalkanoates (PHA) using mixed cultures instead of pure cultures. The main goal in this work was to develop a mathematical model of mixed cultures envisaging the optimisation of PHB production. A relatively simple two-compartments cell model was developed based on experimental observations and other models proposed in the literature. A convenient experimental planing allowed to identify the kinetic parameters and yield coefficients. Experiments were performed with and without ammonia limitation enabling the analysis of PHB formation independently of the cell growth process. The experimental true yields partially confirm the theoretical values proposed in the literature. The final model exhibited high accuracy in describing the process state of most experiments performed, thus opening good perspectives for future model-based optimisation studies.

[1]  S. Jørgensen,et al.  Estimation of kinetic parameters in a structured yeast model using regularisation. , 2001, Journal of biotechnology.

[2]  Mogens Henze,et al.  Yield determination by respirometry - The possible influence of storage under aerobic conditions in activated sludge , 1999 .

[3]  W. Gujer,et al.  Activated sludge model No. 3 , 1995 .

[4]  J J Heijnen,et al.  An integrated metabolic model for the aerobic and denitrifying biological phosphorus removal. , 1997, Biotechnology and bioengineering.

[5]  Ralf Cord-Ruwisch,et al.  The effect of dissolved oxygen on PHB accumulation in activated sludge cultures. , 2003, Biotechnology and bioengineering.

[6]  F. W. Gilcreas,et al.  Standard methods for the examination of water and waste water. , 1966, American journal of public health and the nation's health.

[7]  Kazuyuki Shimizu,et al.  Modeling of the mixed culture and periodic control for PHB production , 2002 .

[8]  H. Chang,et al.  Construction of plasmids, estimation of plasmid stability, and use of stable plasmids for the production of poly(3-hydroxybutyric acid) by recombinant Escherichia coli. , 1994, Journal of biotechnology.

[9]  Mauro Majone,et al.  Aerobic storage under dynamic conditions in activated sludge processes. The state of the art , 1999 .

[10]  M. Loosdrecht,et al.  Production of polyhydroxyalkanoates by mixed microbial cultures , 2003, Bioprocess and biosystems engineering.

[11]  Zhiguo Yuan,et al.  Modeling aerobic carbon oxidation and storage by integrating respirometric, titrimetric, and off‐gas CO2 measurements , 2004, Biotechnology and bioengineering.

[12]  Mauro Majone,et al.  A bulking sludge with high storage response selected under intermittent feeding , 1998 .

[13]  J J Heijnen,et al.  Simultaneous storage and degradation of PHB and glycogen in activated sludge cultures. , 2001, Water research.

[14]  M. V. van Loosdrecht,et al.  Modelling of activated sludge processes with structured biomass. , 2002, Water science and technology : a journal of the International Association on Water Pollution Research.

[15]  H. Chang,et al.  Production of poly(3‐hydroxybutyric acid) by fed‐batch culture of Alcaligenes eutrophus with glucose concentration control , 1994, Biotechnology and bioengineering.

[16]  Mogens Henze,et al.  Wastewater Treatment: Biological and Chemical Processes , 1995 .

[17]  Tomonori Matsuo,et al.  Uptake of Organic Substrates and Accumulation of Polyhydroxyalkanoates Linked with Glycolysis of Intracellular Carbohydrates under Anaerobic Conditions in the Biological Excess Phosphate Removal Processes , 1992 .

[18]  Y. Comeau,et al.  Determination of Poly-β-Hydroxybutyrate and Poly-β-Hydroxyvalerate in Activated Sludge by Gas-Liquid Chromatography , 1988 .

[19]  W. Page,et al.  Growth of Azotobacter vinelandii UWD in Fish Peptone Medium and Simplified Extraction of Poly-β-Hydroxybutyrate , 1993, Applied and environmental microbiology.

[20]  R. M. Lafferty,et al.  A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass , 1978, European journal of applied microbiology and biotechnology.

[21]  G T Daigger,et al.  An assessment of the role of physiological adaptation in the transient response of bacterial cultures , 1982, Biotechnology and bioengineering.

[22]  Y. W. Lee,et al.  Increased PHB productivity by high‐cell‐density fed‐batch culture of Alcaligenes latus, a growth‐associated PHB producer , 1996, Biotechnology and bioengineering.

[23]  M. V. van Loosdrecht,et al.  Storage and degradation of poly-beta-hydroxybutyrate in activated sludge under aerobic conditions. , 2001, Water research.

[24]  J. J. Heijnen,et al.  Poly-β-hydroxybutyrate metabolism in dynamically fed mixed microbial cultures , 2002 .

[25]  Rashmi,et al.  Polyhydroxyalkanoates: an overview. , 2003, Bioresource technology.

[26]  Rui Oliveira,et al.  Optimization of polyhydroxybutyrate production by mixed cultures submitted to aerobic dynamic feeding conditions , 2004, Biotechnology and bioengineering.

[27]  H Shi,et al.  Dynamics and modeling on fermentative production of poly (beta-hydroxybutyric acid) from sugars via lactate by a mixed culture of Lactobacillus delbrueckii and Alcaligenes eutrophus. , 1999, Journal of biotechnology.

[28]  D. Dionisi,et al.  Aerobic storage by activated sludge on real wastewater. , 2001, Water research.

[29]  J J Heijnen,et al.  Kinetic modeling of poly(beta-hydroxybutyrate) production and consumption by Paracoccus pantotrophus under dynamic substrate supply. , 1997, Biotechnology and bioengineering.

[30]  J J Heijnen,et al.  Stoichiometry and kinetics of poly-beta-hydroxybutyrate metabolism in aerobic, slow growing, activated sludge cultures. , 2000, Biotechnology and bioengineering.

[31]  M. Majone,et al.  Comparison of carbon storage under aerobic and anoxic conditions , 1998 .

[32]  M. Loosdrecht,et al.  Substrate flux into storage and growth in relation to activated sludge modeling , 1999 .

[33]  Tomonori Matsuo,et al.  Anaerobic uptake of glutamate and aspartate by enhanced biological phosphorus removal activated sludge , 1998 .