Optimization of Culture Conditions and Characterization of Pectinase from Isolated Bacteria and Application in Juice Clarificationz`

Abstract

Microbial pectinase has high demand in different industries. Pectinase producing bacteria were isolated and purified from soil samples and screened by primary and secondary screening for pectinase production. Out of the 13 bacterial samples, sample 4.1(S4.1) showed consistent enzyme activity. Optimum production of pectinase was high when incubated at 24-hour at 37°C with 2% substrate concentration maintaining pH6. Optimum pectinase activity was observed at temperature 37°C and pH 6. Cu++ was found as activator and Zn++, Ca++ as inhibitors of pectinases. The values for Km and Vmax were found to be 1.181mg/ml and 471.69µM/min respectively. Pectinase was efficiently partially purified by 90% acetone saturation. Specific enzyme activity for partially purified enzyme with acetone was 96870 U/mg and purification fold was 35.41. The utilization of low-cost agro-industrial wastes as substrates has been preferable in pectinase production. The sweet lemon and lemon fruit peel powder was observed good substrate for pectinase production. Furthermore, the yield and clarity of sweet lemon juice increased as the concentration of pectinase increased, indicating its potential use in juice processing. Experimental results suggest pectinases from S4.1 was of high value and can be utilized for fruit juice clarification.