[Studies on characteristics of kinetics and metabolic shift of genetically engineered yeast Pichia pastoris in high-density chemostat cultivation].

Kinetics and metabolic shift of DNA recombinant yeast Pichia pastoris was studied in high-density chemostat cultivation in which the glycerol was used as a limited substrate. The experimental results showed that (1) cell optical density (OD600) of Pichia pastoris was linear with its dry cell weight (DCW) and wet cell weight (WCW), and reached 100 OD600 = 128.3 g WCW/L or 100 OD600 = 22.9 g DCW/L; (2) The relationship of specific growth rate (mu) and residual concentration of glycerol is accorded with Monod Equation, to obtain mu = mumax S/(Ks + S), where mumax = 0.366 h-1, Ks = 0.1823 g/L. The maximum cell yield of glycerol substrate YG = 0.54 g/g, growth maintaining coefficient m = 0.0069 g/(g.h), The cell yield of oxygen YX/O2 = 30.96 g/moL, and the optimum theoretical dilution rate Dm = 0.341 h-1 by parameter inference; (3) With specific growth rate (mu) increasing, glycerol metablic flux shifts linearly from glycogenesis and hexosephosphate pathways to glycolysis and tricarboxylic acid cycle pathways on basis of variations of consumption rate of NH3.H2O and respiratory quotient value.