In this study, the optimum conditions for chitin deacetylase (CDA) production by Aspergillus flavus F1 in solid-state fermentation were investigated via two optimization strategies: classical optimization based on the method of one factor at a time and statistical optimization using response surface methodology. The result of classical optimization showed that corn supplemented with 2% chitin moisturized with mineral salts solution at pH=7 and five days of incubation time were the optimum conditions for increasing CDA production with approximately yield of 219.5 U/g solid substrate. Furthermore, pH, moisture level and inoculum size were systemically evaluated to improve CDA production based on a central composite design using the Design expert 7 software. Based on the enhanced regression model, a maximum predicted CDA enzyme production of 283.8 U/g could be obtained with pH 8, moisture level 1:1 (w/v) and inoculum size 3 ml/10g solid substrate, which contain 1×106 spore/ml. The verification of optimization results and determine accuracy of model showed that the actual response of CDA was 312 U/g, which approximately closed to predicted value 283.89 U/g. The crude extract of CDA was concentrated by sucrose. The results showed that 61% of CDA enzyme was yielded with a purification fold of 1.1 In addition. Then CDA was purified partially by gel filtration chromatography after concentration by sucrose with total enzyme activity and specific activity of 1476 U and 12300 (U/mg protein) respectively. Furthermore, the produced CDA enzyme showed maximum activity in pH ranged from (6-8); in which enzyme activity was 69 U/ ml. however; the enzyme stability has a wide range in acidic and alkaline pH. In addition, the enzyme was maintained its activity at temperatures from 30 to 55C˚. Whereas, the activity was declined in temperature up to 55Cº with a minimum activity (8 U/ml) observed at 80Cº.
An Application of Response Surface Methodology for Optimizing the Production of Chitin Deacetylase Enzyme by Aspergillus Flavus