Metabolic flux-based optimisation of recombinant human interleukin-3 expression by Streptomyces lividans 66

Autor: Keyvan Nowruzi, Ali Elkamel, Murray Moo-Young, Jeno M. Scharer, Douglas Cossar
Rok vydání: 2011
Předmět:
Zdroj: The Canadian Journal of Chemical Engineering. 89:879-888
ISSN: 0008-4034
DOI: 10.1002/cjce.20447
Popis: A metabolic reaction network including carbohydrate and amino acid metabolism in both anabolic and catabolic reactions was developed for Streptomyces lividans. Based on observed nutrient consumption, the fermentation process comprised two stages: amino acid uptake in the first phase followed by ammonia uptake as the nitrogen source in the second phase. Linear programming in combination with eight measured input and output fluxes was employed to determine the flux distribution through the major metabolic pathways in both phases of the batch fermentations. Two different objective functions were defined and examined: optimising biomass production by maximising the specific growth rate and optimising a balanced redox metabolism by minimising excess NAD(P)H production. The flux distributions obtained for the two objectives were very similar. The model predicted specific growth rate agreed well (8–10% difference) with observations. A uniform distribution was assumed for each biomass component and the results showed that, using either objective the biomass composition does not have a significant effect on the internal distribution of fluxes. Un reseau de reactions metaboliques comprenant le metabolisme d'hydrates de carbone et d'acides amines, a la fois dans des reactions anaboliques et cataboliques, a ete developpe pour Streptomyces lividans. Une programmation lineaire en combinaison avec huit flux mesures d'entree et de sortie a ete utilisee pour determiner la distribution de flux de voies metaboliques majeures et lors de fermentations en lots. Deux fonctions objectives differentes ont ete definies et examinees : l'optimisation de la production de biomasse en maximisant le taux de croissance specifique et optimisation d'un metabolisme redox equilibre en reduisant au minimum la production excessive de NAD(P)H. En se basant sur la consommation observee d'elements nutritifs, le processus de fermentation a ete divise en deux etapes. Les distributions de flux obtenues pour les deux objectifs etaient tres similaires. On a suppose une distribution uniforme pour chaque composant de biomasse et les resultats montraient qu'avec l'utilisation de l'un ou de l'autre objectif, la composition de biomasse n'avait pas d'effet significatif sur la distribution interne des flux. © 2011 Canadian Society for Chemical Engineering
Databáze: OpenAIRE