Metabolic control analysis of Aspergillus niger L-arabinose catabolism.
A mathematical model of the L-arabinose/D-xylose catabolic pathway of Aspergillus niger was constructed based on the kinetic properties of the enzymes. For this purpose L-arabinose reductase, L-arabitol dehydrogenase and D-xylose reductase were purified using dye-affinity chromatography, and their kinetic properties were characterized. For the other enzymes of the pathway the kinetic data were available from the literature. The metabolic model was used to analyze flux and metabolite concentration control of the L-arabinose catabolic pathway. The model demonstrated that flux control does not reside at the enzyme following the intermediate with the highest concentration, L-arabitol, but is distributed over the first three steps in the pathway, preceding and following L-arabitol. Flux control appeared to be strongly dependent on the intracellular L-arabinose concentration. At 5 mM intracellular L-arabinose, a level that resulted in realistic intermediate concentrations in the model, flux control coefficients for L-arabinose reductase, L-arabitol dehydrogenase and L-xylulose reductase were 0.68, 0.17 and 0.14, respectively. The analysis can be used as a guide to identify targets for metabolic engineering aiming at either flux or metabolite level optimization of the L-arabinose catabolic pathway of A. niger. Faster L-arabinose utilization may enhance utilization of readily available organic waste containing hemicelluloses to be converted into industrially interesting metabolites or valuable enzymes or proteins.
de Groot
Marco J L
MJ
Fungal Genomics and Section of Molecular Genetics of Industrial Microorganisms, Wageningen University, Dreijenlaan 2, The Netherlands. mjl_degroot@kimarco.nl
Prathumpai
Wai
W
Visser
Jaap
J
Ruijter
George J G
GJ
D000449:Aldehyde Reductase / Q000302:isolation & purification / Q000378:metabolism; D001089:Arabinose / Q000378:metabolism*; D001234:Aspergillus niger / Q000201:enzymology / Q000378:metabolism*; D001709:Biotechnology; D007700:Kinetics; D008954:Models, Biological; D013401:Sugar Alcohol Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism; D014994:Xylose / Q000378:metabolism
D014994:Xylose; D001089:Arabinose; D013401:Sugar Alcohol Dehydrogenases; D000449:Aldehyde Reductase