Pectinase Production of Aspergillus sp. VTM5 Through Solid State Fermentation Using Coffee Pulp Substrate and Its Purification

Abstract

Hydrolytic Pectinase is an important enzyme and is widely used in several industries. In the previous investigation, this enzyme could be produced by Aspergillus sp. VTM5 under solid-state fermentation and the coffee pulp waste substrates were used. The specific activity of about 0.4 U/ml of pectinase was detected when 40gr coffee pulp water-saturated inoculated with 1.2x10 7 spores of Aspergillus sp. VTM5 was incubated at 30 °C for 168 h, and without any nutrients added. Against 0.5% alkaline extract of pomelo pulp substrate in 20 Mm acetate buffer pH 5, pectinase activity increases up to 1.03 U/ml, and the purity 3.5 times, after dialyzed using 12-14 kDa, pores cellulose membrane tube higher than previously step. Open column purification using Fractogel EMD SO3-(S) cation exchange chromatography, pectinase was eluted at 0.45 M NaCl. The pectinase activity significantly increased to 0.295 U/ml with 12.37-fold and 17.04% yield. Based on these results, it can be proved that coffee pulp waste is suitable as a substrate for pectinase production by Aspergillus sp. VTM5. Further research to analyze the properties and characterize the stability of pectinase against temperature, pH, the effect of heavy metals, and molecular analysis is needed.