The present study deals with the coproduction of laccase and pectinase enzymes through solid state fermentation using mixed fungal culture of Trametes hirsuta and Phanerochaete sp., to minimize the cost and time of the process. Substrates selected for the enzyme production were wheat bran, pulse husk and mustard peel. To get optimum yield of laccase and pectinase in a single fermenter, EVOP factorial design technique with factors like pH, incubation temperature and substrates ratio have been explored. At search level of EVOP, outcomes of the one factor at a time has been considered as incubation temperature 30 °C for 7 days at pH 5 with wheat bran: pulse husk: mustard peel ratio of 2:2:1 (w/w/w) and yield of laccase and pectinase was found as 78.1 U/gds and 105.0 U/gds respectively. In first setup of EVOP factorial both laccase and pectinase activities were not found optimum in same set of experiments; therefore, on the basis of its decision making steps, second set of experiment was performed by taking decisions of first set of EVOP as search level. Optimum yield of laccase and pectinase was achieved about 250 U/gds and 247 U/gds respectively at 34 °C, pH 4.5 with 1.75:1.75:1.5 (w/w/w) wheatbran: pulse husk: mustard peel as substrate, which was 2-3 times higher than the outcomes of one factor at a time method.
In order to achieve high yield of fungal protease in a very cost effective way and to meet its increased market demand, current study deals with the screening of various agro-wastes as carbon source for the production of protease from Rhizopus oryzae (SN5) / NCIM-1447 under solid state fermentation. Substrates and culture parameters such as wheat bran, soybean meal, black-gram husk, rice husk, mixture of wheat bran, soybean meal, nitrogen sources, pH, temperature and incubation time were first optimized with one factor at time strategy and then EVOP factorial and yield of alkaline protease was achieved 412.8 U/gds at 28oC and pH=6 after 72 hours of fermentation taking wheat bran and soybean as a substrate in 4:1 ratio. Further Artificial Neural Networks (ANN), was trained with data of EVOP and yield of protease was enhanced up to 422.6 U/gds with wheat bran: soyabean in ratio of 70:30, pH 6.2 at 30°C. The evolved process and Rhizopus oryzae (SN5)/NCIM-1447 strain would be promising for protease production at industrial scale at low cost.
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