含离子液体介质中重组β-D-葡萄糖醛酸苷酶催化合成单葡萄糖醛酸基甘草次酸的研究-生物化工专业论文.docxVIP

石河子大学硕士学位论文Abs 石河子大学硕士学位论文 Abstract IV IV %. When 10% (v/v) ionic liquid/buffer mixed solvents were used as the reaction media, all the conversions of GL were increased to 2~3 fold of that in pure ionic liquids. Of the six media tested, 10% (v/v) [BMIM]PF6/buffer biphasic system and 10% (v/v) [EMIM]BF4/buffer monophasic system showed higher conversions, 57.13% and 54.23%, respectively, much more than that in buffer system. Therefore, they had been chosen as the suitable ionic liquid-containing media. After preliminary optimization, the initial reaction rate, the conversion of GL, the GAMG yield and the chemical bond selectivity (Scb) reached 2.9 mMh-1, 71.71%, 72.62% and 98.76% in the 40%(v/v) [BMIM]PF6/buffer biphasic system, respectively. Catalytical characteristic of β-D-glucuronidase in [BMIM]PF6/buffer biphasic system: β-D-glucuronidase showed high catalytical efficiency for hydrolysis of GL to GAMG in [BMIM]PF6/buffer biphasic system. Many factors showed influence on the initial reaction rate and chemical bond selectivity. The optimized reaction conditions in [BMIM]PF6/buffer biphasic system were [BMIM]PF6 content 40%, pH 6.0, temperature 50 ℃, substrate concentration 6 mM and shaking speed 200 rpm. Under these optimal conditions the initial reaction rate, the GAMG yield and the Scb reached 3.15 mMh-1, 74.36% and 98.12%, respectively. The desired product was distributed in the upper buffer phase in the [BMIM]PF6/buffer biphasic system, while the by-product glycyrrhetinic acid (GA) distributed in the lower layer of ionic liquid, so GAMG and GA were separated automaticly. The recyclability of [BMIM]PF6 was also studied and found to be reusable for 5 batches still with high recovery percentage (≥ 92%). Additionally, higher Vmax, lower apparent Km and Ea were achieved in [BMIM]PF6/buffer biphasic system, they are 3.14 mMh-1, 1.21 mM and 25.97 kJmol-1, respectively. Catalytical characteristic of β-D-glucuronidase in [EMIM]BF4/buffer monophasic system: T