A kinetic study of reuterin production by Lactobacillus reuteri PRO 137 in resting cells

Abstract This work deals with the production of reuterin by Lactobacillus reuteri PRO 137 in a two-step batch fermentation process. First, L. reuteri cells were grown anaerobically in a glucose-containing media, and in a second stage reuterin was produced by the starved resting cells in glycerol-containing media. In order to maximize biomass production, the influence of temperature (31–43 °C) and pH (5.0–6.5) was evaluated. A simple model has been proposed to describe the microbial growth and substrate consumption. The values of the kinetic constant for growth rate, yield of biomass on substrate and maximum biomass concentration indicate that the optimum operating conditions were 40 °C and pH 5.5. Biomass produced in those growth conditions was harvested, washed and resuspended in a medium containing glycerol as the only carbon source for reuterin production by resting cells. Reuterin production was evaluated for several biomass (2 and 5 CDW g/L) and glycerol (150–300 mM) concentrations. A model describing the kinetics of the production process has been proposed. The model allows to describe simultaneously the concentrations of viable cells, glycerol, reuterin and 1,3-propanediol. A multi-response nonlinear parameter estimation procedure using the Marquardt algorithm and numerical integration of the differential equations by Runge-Kutta method was used to obtain the values of the fitting parameters.

[1]  P. Neysens,et al.  Modeling Growth and Bacteriocin Production by Lactobacillus amylovorus DCE 471 in Response to Temperature and pH Values Used for Sourdough Fermentations , 2002, Applied and Environmental Microbiology.

[2]  J. Thibault,et al.  Lactobacillus helveticus growth and lactic acid production during pH-controlled batch cultures in whey permeate/yeast extract medium. Part II: kinetic modeling and model validation , 2002 .

[3]  M. Rogosa,et al.  A MEDIUM FOR THE CULTIVATION OF LACTOBACILLI , 1960 .

[4]  K. Venkatesh,et al.  Structured Model for Batch Culture Growth of Lactobacillus bulgaricus , 1997 .

[5]  L. Axelsson,et al.  In Vitro Studies on Reuterin Synthesis by Lactobacillus reuteri , 2009 .

[6]  A. Amrane,et al.  Mathematical model for lactic acid production from lactose in batch culture: Model development and simulation , 1994 .

[7]  J. Debevere,et al.  The effect of Reuterin on Listeria monocytogenes and Escherichia coli O157:H7 in milk and cottage cheese. , 1998, Journal of food protection.

[8]  M. Jakobsen,et al.  Characterisation and modelling of oscillatory behaviour related to reuterin production by Lactobacillus reuteri. , 2002, International journal of food microbiology.

[9]  D. Marquardt An Algorithm for Least-Squares Estimation of Nonlinear Parameters , 1963 .

[10]  Janneke Krooneman,et al.  Anaerobic Conversion of Lactic Acid to Acetic Acid and 1,2-Propanediol by Lactobacillus buchneri , 2001, Applied and Environmental Microbiology.

[11]  L. Axelsson,et al.  In VitroStudies on Reuterin Synthesis byLactobacillus reuteri , 1989 .

[12]  M. K. Yunmbam,et al.  The in vitro efficacy of reuterin on the culture and bloodstream forms of Trypanosoma brucei brucei. , 1992, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.

[13]  W. Dobrogosz,et al.  Chemical characterization of an antimicrobial substance produced by Lactobacillus reuteri , 1989, Antimicrobial Agents and Chemotherapy.

[14]  L. Axelsson,et al.  Utilization of Glycerol as a Hydrogen Acceptor by Lactobacillus reuteri: Purification of 1,3-Propanediol:NAD+ Oxidoreductase , 1990, Applied and environmental microbiology.

[15]  R. Cachon,et al.  Generalized model of the effect of pH on lactate fermentation and citrate bioconversion in Lactococcus lactis ssp. Lactis biovar. diacetylactis , 1994, Applied Microbiology and Biotechnology.

[16]  I. Casas,et al.  Production and isolation of reuterin, a growth inhibitor produced by Lactobacillus reuteri , 1988, Antimicrobial Agents and Chemotherapy.

[17]  J. Arqués,et al.  Antimicrobial activity of reuterin in combination with nisin against food-borne pathogens. , 2004, International journal of food microbiology.

[18]  J. Arqués,et al.  Synergistic effect of reuterin in combination with nisin on Staphylococcus aureus in milk , 2004 .

[19]  P. Neysens,et al.  Effect of sodium chloride on growth and bacteriocin production by Lactobacillus amylovorus DCE 471. , 2003, International journal of food microbiology.

[20]  Q. Lüthi-Peng,et al.  Production and stability of 3-hydroxypropionaldehyde in Lactobacillus reuteri , 2002, Applied Microbiology and Biotechnology.

[21]  Q. Lüthi-Peng,et al.  Effect of glucose on glycerol bioconversion by Lactobacillus reuteri , 2002, Applied Microbiology and Biotechnology.

[22]  J. Debevere,et al.  Characterization of growth and metabolite production of Lactobacillus reuteri during glucose/glycerol cofermentation in batch and continuous cultures , 1998, Biotechnology Letters.

[23]  J. Arqués,et al.  Reuterin production by lactobacilli isolated from pig faeces and evaluation of probiotic traits , 2003, Letters in applied microbiology.

[24]  Jules Thibault,et al.  Lactobacillus helveticus growth and lactic acid production during pH-controlled batch cultures in whey permeate/yeast extract medium. Part I. multiple factor kinetic analysis , 2002 .

[25]  K. L. Smiley,et al.  METABOLISM OF GLYCEROL BY AN ACROLEIN-FORMING LACTO-BACILLUS , 1960, Journal of bacteriology.

[26]  J. Rincón,et al.  Kinetics of Lactic Acid Fermentation by Lactobacillus delbrueckii Grown on Beet Molasses , 1997 .

[27]  N. Sauvageot,et al.  Glycerol metabolism in Lactobacillus collinoides: production of 3-hydroxypropionaldehyde, a precursor of acrolein. , 2000, International journal of food microbiology.

[28]  M. Çakmakci,et al.  Effect of reuterin produced by Lactobacillus reuteri on the surface of sausages to inhibit the growth of Listeria monocytogenes and Salmonella spp. , 2002, Die Nahrung.

[29]  M. Jakobsen,et al.  Application of reuterin produced by Lactobacillus reuteri 12002 for meat decontamination and preservation. , 1999, Journal of food protection.

[30]  L. Axelsson,et al.  Production of a Broad Spectrum Antimicrobial Substance by Lactobacillus reuteri , 1989 .