title: NADH-Regulated metabolic model for growth of Methylosinus trichosporium OB3b. Model presentation, parameter estimation, and model validation.
abstract: A biochemical model is presented that describes growth of Methylosinus trichosporium OB3b on methane. The model, which was developed to compare strategies to alleviate NADH limitation resulting from cometabolic contaminant conversion, includes (1) catabolism of methane via methanol, formaldehyde, and formate to carbon dioxide; (2) growth as formaldehyde assimilation; and (3) storage material (poly-beta-hydroxybutyric acid, PHB) metabolism. To integrate the three processes, the cofactor NADH is used as central intermediate and controlling factor-instead of the commonly applied energy carrier ATP. This way a stable and well-regulated growth model is obtained that gives a realistic description of a variety of steady-state and transient-state experimental data. An analysis of the cells' physiological properties is given to illustrate the applicability of the model. Steady-state model calculations showed that in strain OB3b flux control is located primarily at the first enzyme of the metabolic pathway. Since no adaptation in V(MAX) values is necessary to describe growth at different dilution rates, the organism seems to have a "rigid enzyme system", the activity of which is not regulated in response to continued growth at low rates. During transient periods of excess carbon and energy source availability, PHB is found to accumulate, serving as a sink for transiently available excess reducing power.
authors.affiliation: Chemical Engineering and Biochemistry Departments, University of Groningen, NL-9747 AG Groningen, The Netherlands.
authors.forename: E M
authors.initials: EM
authors.lastname: Sipkema
authors.forename: W
authors.initials: W
authors.lastname: de Koning
authors.forename: K J
authors.initials: KJ
authors.lastname: Ganzeveld
authors.forename: D B
authors.initials: DB
authors.lastname: Janssen
authors.forename: A A
authors.initials: AA
authors.lastname: Beenackers
mesh_terms: D001709:Biotechnology / Q000379:methods; D003198:Computer Simulation; D003470:Culture Media; D005557:Formaldehyde / Q000378:metabolism; D005561:Formates / Q000378:metabolism; D006885:Hydroxybutyrates / Q000378:metabolism; D008697:Methane / Q000378:metabolism; D000432:Methanol / Q000378:metabolism; D020602:Methylosinus trichosporium / Q000254:growth & development* / Q000378:metabolism*; D008954:Models, Biological; D009243:NAD / Q000378:metabolism*; D045542:PQQ Cofactor; D011091:Polyesters / Q000378:metabolism; D015363:Quinolones / Q000378:metabolism; D011809:Quinones / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D000094122:Polyhydroxybutyrates
chemical_list: D003470:Culture Media; D005557:Formaldehyde; D005561:Formates; D006885:Hydroxybutyrates; D008697:Methane; D000432:Methanol; D009243:NAD; D045542:PQQ Cofactor; D011091:Polyesters; D015363:Quinolones; D011809:Quinones; C030544:formic acid; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates