Growth kinetics of Escherichia coli with galactose and several other sugars in carbon-limited chemostat culture.

Kinetic models for microbial growth describe the specific growth rate (mu) as a function of the concentration of the growth-limiting nutrient (s) and a set of parameters. A typical example is the model proposed by Monod, where mu is related to s using substrate affinity (Ks) and the maximum specific growth rate (mu max). The preferred method to determine such parameters is to grow microorganisms in continuous culture and to measure the concentration of the growth-limiting substrate as a function of the dilution rate. However, owing to the lack of analytical methods to quantify sugars in the microgram per litre range, it has not been possible to investigate the growth kinetics of Escherichia coli in chemostat culture. Using an HPLC method able to determine steady-state concentrations of reducing sugars, we previously have shown that the Monod model adequately describes glucose-limited growth of E. coli ML30. This has not been confirmed for any other sugar. Therefore, we carried out a similar study with galactose and found steady-state concentrations between 18 and 840 micrograms.L-1 for dilution rates between 0.2 and 0.8.h-1, respectively. With these data the parameters of several models giving the specific growth rate as a function of the substrate concentration were estimated by nonlinear parameter estimation, and subsequently, the models were evaluated statistically. From all equations tested, the Monod model described the data best. The parameters for galactose utilisation were mu max = 0.75.h-1 and Ks = 67 micrograms.L-1. The results indicated that accurate Ks values can be estimated from a limited set of steady-state data when employing mu max measured during balanced growth in batch culture. This simplified procedure was applied for maltose, ribose, and fructose. For growth of E. coli with these sugars, mu max and Ks were for maltose 0.87.h-1, 100 micrograms.L-1; for ribose 0.57.h-1, 132 micrograms.L-1, and for fructose 0.70.h-1, 125 micrograms.L-1.

[1]  K. van Dam,et al.  Effect of concentration of substrates and products on the growth of Klebsiella pneumoniae in chemostat cultures. , 1989, Biochimica et biophysica acta.

[2]  H. Kornberg,et al.  Uptake of galactose into Escherichia coli by facilitated diffusion. , 1976, Journal of general microbiology.

[3]  D. Dykhuizen,et al.  An Experimental Model: Bacterial Specialists and Generalists Competing in Chemostats , 1980 .

[4]  T. Egli,et al.  Is Escherichia coli growing in glucose-limited chemostat culture able to utilize other sugars without lag? , 1995, Microbiology.

[5]  T. Egli,et al.  Growth Kinetics of Suspended Microbial Cells: From Single-Substrate-Controlled Growth to Mixed-Substrate Kinetics , 1998, Microbiology and Molecular Biology Reviews.

[6]  T. Ferenci,et al.  Derepression of LamB protein facilitates outer membrane permeation of carbohydrates into Escherichia coli under conditions of nutrient stress , 1993, Journal of bacteriology.

[7]  F. Quiocho,et al.  Preliminary crystallographic data of receptors for transport and chemotaxis in Escherichia coli: D-galactose and maltose-binding proteins. , 1979, Journal of molecular biology.

[8]  Winfried Boos,et al.  Maltose Transport in Escherichia coli K12 , 1976 .

[9]  D. Button Biochemical Basis for Whole-Cell Uptake Kinetics: Specific Affinity, Oligotrophic Capacity, and the Meaning of the Michaelis Constant , 1991, Applied and environmental microbiology.

[10]  H. Wiesmeyer,et al.  Regulation of ribose metabolism in Escherichia coli. I. The ribose catabolic pathway. , 1970, Biochimica et biophysica acta.

[11]  K. Hellingwerf,et al.  Thermodynamics of growth. Non-equilibrium thermodynamics of bacterial growth. The phenomenological and the mosaic approach. , 1982, Biochimica et biophysica acta.

[12]  M. Höfle,et al.  Long-Term Changes in Chemostat Cultures of Cytophaga johnsonae , 1983, Applied and environmental microbiology.

[13]  T. Egli,et al.  The growth of Escherichia coli in glucose-limited chemostat cultures: a re-examination of the kinetics. , 1994, Biochimica et biophysica acta.

[14]  J. A. Robinson,et al.  Determining microbial kinetic parameters using nonlinear regression analysis. Advantages and limitations in microbial ecology , 1985 .

[15]  B. Rotman,et al.  GALACTOSE TRANSPORT IN ESCHERICHIA COLI. THE MECHANISM UNDERLYING THE RETENTION OF INTRACELLULAR GALACTOSE. , 1964, The Journal of biological chemistry.

[16]  F. F. Blackman Optima and Limiting Factors , 1905 .

[17]  D E Koshland,et al.  Properties of the galactose binding protein of Salmonella typhimurium and Escherichia coli. , 1977, Biochemistry.

[18]  Growth kinetics of Escherichia coli with mixtures of sugars , 1994 .

[19]  H. Jannasch Competitive elimination of Enterobacteriaceae from seawater. , 1968, Applied microbiology.

[20]  O. Richter Parameter estimation in ecology , 1990 .

[21]  D. E. Contois Kinetics of bacterial growth: relationship between population density and specific growth rate of continuous cultures. , 1959, Journal of general microbiology.

[22]  A. G. Marr,et al.  Effect of Nutrient Concentration on the Growth of Escherichia coli , 1971, Journal of bacteriology.

[23]  J. Monod,et al.  Recherches sur la croissance des cultures bactériennes , 1942 .

[24]  R. K. Finn,et al.  Equations of substrate‐limited growth: The case for blackman kinetics , 1973, Biotechnology and bioengineering.

[25]  C. Furlong,et al.  Purification and properties of a ribose-binding protein from Escherichia coli. , 1974, The Journal of biological chemistry.

[26]  Christon J. Hurst,et al.  Modeling the metabolic and physiologic activities of microorganisms , 1992 .

[27]  D. Wilson The regulation and properties of the galactose transport system in Escherichia coli K12. , 1974, The Journal of biological chemistry.

[28]  M Rutgers,et al.  Establishment of the steady state in glucose-limited chemostat cultures of Klebsiella pneumoniae. , 1987, Journal of general microbiology.

[29]  Sebastiaan A.L.M. Kooijman,et al.  Microbial growth dynamics on the basis of individual budgets. , 1991 .

[30]  R. Harvey Metabolic Regulation in Glucose-Limited Chemostat Cultures of Escherichia coli , 1970, Journal of bacteriology.

[31]  Further studies on the binding of maltose to the maltose-binding protein of Escherichia coli. , 1976, European journal of biochemistry.