Modeling of the biotransformation of crotonobetaine into L-(-)-carnitine by Escherichia coli strains.
A simple unstructured model, which includes carbon source as the limiting and essential substrate and oxygen as an enhancing substrate for cell growth, has been implemented to depict cell population evolution of two Escherichia coli strains and the expression of their trimethylammonium metabolism in batch and continuous reactors. Although the model is applied to represent the trans-crotonobetaine to L-(-)-carnitine biotransformation, it is also useful for understanding the complete metabolic flow of trimethylammonium compounds in E. coli. Cell growth and biotransformation were studied in both anaerobic and aerobic conditions. For this reason we derived equations to modify the specific growth rate, mu, and the cell yield on the carbon source (glycerol), Y(xg), as oxygen increased the rate of growth. Inhibition functions representing an excess of the glycerol and oxygen were included to depict cell evolution during extreme conditions. As a result, the model fitted experimental data for various growth conditions, including different carbon source concentrations, initial oxygen levels, and the existence of a certain degree of cell death. Moreover, the production of enzymes involved within the E. coli trimethylammonium metabolism and related to trans-crotonobetaine biotransformation was also modeled as a function of both the cell and oxygen concentrations within the system. The model describes all the activities of the different enzymes within the transformed and wild strains, able to produce L-(-)-carnitine from trans-crotonobetaine under both anaerobic and aerobic conditions. Crotonobetaine reductase inhibition by either oxygen or the addition of fumarate as well as its non-reversible catalytic action was taken into consideration. The proposed model was useful for describing the whole set of variables under both growing and resting conditions. Both E. coli strains within membrane high-density reactors were well represented by the model as results matched the experimental data.
Canovas
M
M
Departamento de Bioquímica y Biología Molecular B e Inmunología, Facultad de Química, Universidad de Murcia, Apdo. Correos 4021, 30100 Murcia, Spain.
Maiquez
J R
JR
Obón
J M
JM
Iborra
J L
JL
D000465:Algorithms; D001622:Betaine / Q000031:analogs & derivatives* / Q000378:metabolism*; D019149:Bioreactors; D002331:Carnitine / Q000032:analysis* / Q000096:biosynthesis*; D003198:Computer Simulation; D004926:Escherichia coli / Q000254:growth & development* / Q000378:metabolism; D005650:Fumarates / Q000378:metabolism; D008566:Membranes / Q000502:physiology; D008954:Models, Biological; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors; D010088:Oxidoreductases / Q000037:antagonists & inhibitors; D013997:Time Factors
D005650:Fumarates; D009097:Multienzyme Complexes; D001622:Betaine; C032005:fumaric acid; C025237:crotonic acid betaine; D010088:Oxidoreductases; C067153:crotonobetaine reductase; D002331:Carnitine