Effect of Hydrophilic Ionic Liquid [EMIM]BF4 on the Catalytic Activity of Immobilized β-Glucuronidase

doi:10.7503/cjcu20121082

Abstract

Abstract:

To improve hydrolysis of glycyrrhizin(GL) to glycyrrhetinic acid monoglucuronide(GAMG) possessing best biological function, Penicillium purpurogenum Li-3 strains highly expressing β-glucuronidase was screened by our workgroup in the initial research period. This enzyme was capable of promoting biotransformation process of GL to GAMG. With chitosan as carrier, immobilized β-glucuronidase was prepared by adsorption-crosslink technology in this work. With the purpose of studying the effect of hydrophilic ionic liquid BF₄ on catalytic activity of immobilized β-glucuronidase, the [EMIM]BF₄/buffer homogeneous system was used as bio-catalytic medium for immobilized β-glucuronidase to catalyze hydrolysis of GL to GAMG. The results showed that under the conditions of 16%(volume fraction)[EMIM]BF₄, pH value of 5.4, the reaction temperature of 50℃ and the shaker rotate speed of 200 r/min, the enzyme activity could reach its maximum, and obviously higher than the highest enzyme activity in pure buffer medium system. After repeated recycling experiment, compared with that in pure buffer medium system, the immobilized β-glucuronidase showed excellent stability to repeated use in containing [EMIM]BF₄ homogeneous medium system. The appa-rent kinetic parameters and activation energy data showed that the affinity between β-glucuronidase and its substance was enhanced by [EMIM]BF₄ in biocatalytic medium, and the enzyme-substrate in the transition state was effectively stabilized, and the activation energy of the catalytic reaction was lowered, so that immobilized β-glucuronidase showed higher catalytic activity. This study provided the basis for design and application of new clean biological reactors and technology to improve the efficiency of enzymatic hydrolysis of GL to GAMG.

Key words: Hydrophilic ionic liquid, BF₄, Immobilized β-glucuronidase, Catalytic activity, Glycyrrhizin, Glycyrrhetinic acid monoglucuronide

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CAO Hong, SUN Qian, LI Chun, LI Kai-Xiong, LI Jun. Effect of Hydrophilic Ionic Liquid [EMIM]BF₄ on the Catalytic Activity of Immobilized β-Glucuronidase[J]. Chem. J. Chinese Universities, 2013, 34(8): 1873.

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Effect of Hydrophilic Ionic Liquid [EMIM]BF₄ on the Catalytic Activity of Immobilized β-Glucuronidase

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