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The optimization of the various process parameters including nutritional and physiochemical factors was carried out for production of pectinase using from mutant strain of Bacillus subtilis using Taguchi methodology. In the entire optimization study, mutant UV-20-11/NTG-1-5/EMS-1-2 was used. The experimental work was performed using shake flask experiments. In this study, ten factors including, orange peel, ammonium sulfate, magnesium sulfate, phosphate ion, sodium chloride, inoculum percentage, temperature, pH, agitation speed, and incubation time were considered for their effect on pectinase production. It was evident from Taguchi methodology that agitation speed contributed the maximum impact (26.51%) on pectinase productivity followed by ammonium sulfate (13.77%), incubation time (12.77%), temperature (12.36%), pH (V%), orange peel (9.64%), inoculum percentage (5.61%), and sodium chloride (5.467%). The results showed that a higher level of pectinase productivity (630.58 U) was achieved with orange peel (5 g/L), ammonium sulfate (1.5 g/L), magnesium sulfate (0.5 g/L), phosphate ion (0.5 g/L), sodium chloride (0.5 g/L), inoculum percentage (1%), temperature (37 oC), pH (7), agitation speed (140 rpm), and incubation time (96 h).