The objective of this study is to optimize the different parameters to carry out analysis of fatty acids. A kinetic was observed for first order enzymatic hydrolysis of flax seed methyl ester was carried out by using Rhizomucor michei. In this study the analysis of hydrolysis was carried out by varying the temperature (30-40oC) and enzyme load (2-5%). The optimal condition were found to temperature 50oC, 6h reaction time, buffer to flax seed methyl ester ratio 1.5:1(v/w) and 4% enzyme load to achieve a maximum hydrolysis conversion of 97.56%. The effect of temperature on the reaction rate constant and equilibrium constant has been determined using Arrhenius equation. The heat of reaction was found 14.516 KJ/mol. Taguchi's design of experiment L16 and L9 orthogonal array was performed to optimize hydrolysis reaction conditions. Rate of reaction, effect of temperature, enzyme modifier, pH and oil to buffer ratio were considered as a primary influencing parameters which effects the percentage of hydrolysis and fatty acid formed. From the analysis of variance, the influencing parameters on production of fatty acid were reaction time and enzyme modifier. The predicted conversion was found in good rectification with experimental values having R2=0.9945 and R2=0.983. Maximum fatty acid formed was 98.76% from methyl ester and 98.92% from oil.
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