Effect of aprotic solvents on the enzymatic activity of lipase in AOT reverse micelles |
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Authors: | Muhammad Moniruzzaman Yoshishige Hayashi Md. Mahabubur Rahman Talukder Eiji Saito Takuya Kawanishi |
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Affiliation: | aDivision of Material Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma machi, Kanazawa 920-1192, Japan bInstitute of Chemical and Engineering Sciences, 1 Pesek Road, Jurong Island, Singapore 627833, Singapore |
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Abstract: | The modification of reverse micellar systems composed of AOT, isooctane, water by the addition of aprotic solvents has been performed. The impact of this change on the activity, stability and kinetics of solubilized Chromobacterium viscosum lipase (glycerol-ester hydrolase, EC 3.1.1.3) was investigated. Of seven aprotic solvents tested, dimethyl sulfoxide (DMSO) was found to be most effective. It was found that lipase activity was enhanced by optimizing some relevant parameters, such as water–AOT molar ratio (W0), buffer pH and surfactant concentration. A kinetic model that considers the free substrate in equilibrium with the substrate adsorbed on the micellar surface was successfully used to deduce some kinetic parameters (Vmax, Km and Kad), and the values of Km and Kad were significantly reduced by the presence of DMSO. Higher lipase stability was found in AOT reverse micelles with DMSO compared with that in simple AOT systems with half-life of 125 and 33 days, respectively. Fluorescence spectroscopy and Fourier transform infrared spectroscopy (FT-IR) were used to elucidate the effects of DMSO on the properties of AOT reverse micelles. |
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Keywords: | Reverse micelles Lipase Hydrolysis DMSO Enzyme stability and kinetics |
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