The effect of microbiological parameters, Viz., slant age, inoculum age and amount of inoculum were studied using response surface methodology for the production of pectolytic enzymes, polymethylgalacturonase, polygalacturonase and pectinlyase. The central composite design was used to determine the individual optimum values. Individual optimum values of slant age, inoculum age and amount of inoculum were 49.5 h, 52.3 h and 11.2%, respectively, for polymethylgalacturonase; 62.4 h, 60.5 h and 14.4%, respectively, for polygalacturonase, and 78.3 h, 52.5 h and 1314%, respectively, for pectinlyase. In order to develop a two-stage inoculum possessing the best conditions for the maximum production of all three enzymes, a multiresponse analysis was carried out using a generalized distance approach. The microbiological parameters were optimized simultaneously for maximum production of enzymes. The simultaneous optimal values of slant age, inoculum age and amount of inoculum were 81.4 h, 66.7 h and 15.8%, respectively, for all the three enzymes. After optimization, the fermentation time was reduced from 144 to 120 h.