Abstract
Several methods have been demonstrated in the recovery and purification of cutinase; however, hydrophobic interaction membrane chromatography (HIMC) has yet to be implemented for this purpose. In this study, two factors, that is, buffer pH and ammonium sulphate concentration for HIMC operation, were optimized using response surface methodology (RSM), aiming to obtain the highest cutinase purification factor and recovery possible. The experimental design used was a central composite design (CCD) comprised of 13 experimental runs. The analysis of variance (ANOVA) indicated that a quadratic model best represented the system. Based on the R 2 values, it can be concluded that around 90% of the variability in the system can be explained by the fitted models. The optimized HIMC conditions were buffer pH 6.0 and 1.3 M ammonium sulphate, which was expected to provide 27-fold cutinase purification factor and 85% recovery. However, the actual values deviated from the predicted values with 13% and 18% error, respectively. The statistical analysis showed that only the ammonium sulphate concentration had a great influence on cutinase purification factor. Meanwhile, for cutinase recovery, both parameters were important in determining the optimal condition for cutinase adsorption and desorption to the chromatographic media.
Published Version
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