Response surface methodology as optimization strategy for asymmetric bioreduction of (4S)-(+)-carvone by Cryptococcus gastricus

Response surface methodology was applied in optimizing the asymmetric bioreduction of (4S)-(+)-carvone to dihydrocarvone (with low incidence of unsought side reactions) by using whole-cells of Cryptococcus gastricus. A factorial design (25) including five independent variables was performed: X1 = incubation time; X2 = pH; X3 = amount of whole-cells; X4 = concentration of (4S)-(+)-carvone; X5 = concentration of cofactor-recycling system. The utilization of glucose and glycerol as cofactor-recycling systems was checked. On the basis of the results of factorial design, three independent variables (X1, X3 and X4) out of five were further selected for performing a central composite design (CCD). and second order polynomial equations obtained by CCD were used to select the optimal values of independent variables in order to maximize the bioreduction yield of (4S)-(+)-carvone and, at the same time, to minimize the occurrence of side reactions (i.e. further reduction of dihydrocarvone to dihydrocarveol).