摘要异槲皮苷具有多种药理活性，由于在植物中含量极低导致提取相当困难，而化学法合成成本较高，从而限制了实际应用。本文以价格低廉的芦丁作为底物，用柚皮苷酶作为催化剂，采用酶催化水解法制备异槲皮苷，不使用任何有害化学品和溶剂，但反应效率低，耗时长，酶成本高，且难以重复利用。因此，急需一种绿色、低耗、高效的生物过程来制备异槲皮苷。80807

（1）建立柚皮苷酶催化芦丁水解制备异槲皮苷的工艺。考察底物浓度、酶浓度、温度和pH值对酶催化芦丁水解制备异槲皮苷的影响，结果表明：pH 7的柠檬酸-磷酸氢二钠缓冲液配制的浓度2。0 g/L的芦丁溶液和浓度20 g/L的柚皮苷酶，在50 ℃下反应1 h，异槲皮苷的得率为86。64%。

（2）采用纳米粒子固定化的方法制备固定化柚皮苷酶，并优化制备工艺。对载体和酶量的比值、温度和时间等因素进行考察，结果表明：酶浓度为15 g/L，石墨烯：酶液=4:1（m/v） 时，在30 ℃下处理5 h，固定化效率达到最大值，为28。37%。

（3）固定化柚皮苷酶催化芦丁水解合成异槲皮苷的工艺优化。在常规反应器中，固定化酶催化芦丁水解制备异槲皮苷的最优条件：2 g/L的芦丁溶液与20 g/L柚皮苷酶溶液，在50 ℃混合反应1 h，异槲皮苷得率为86。64%。

（4）微反应器中固定化柚皮苷酶催化芦丁水解合成异槲皮苷的工艺优化。选择十二烷基苯磺酸钠作为表面活性剂，当底物浓度为0。05 g/L、温度50 ℃、流速4 μL/min、溶液pH为7时，异槲皮苷的得率高达92。78%。

毕业论文关键词：纳米粒子；固定化酶；芦丁；异槲皮苷；微反应器

AbstractIsoquercitrin has many pharmacological activities, but it is difficult to extract from plants, while chemical synthesis of isoquercitrin is expensive, which limits the development of isoquercitrin。 Isoquercitrin was synthesized by the naringinase-catalyzed selective hydrolysis of rutin, without using any harmful chemicals and solvents。 However, the reaction time is still long, the cost of enzyme is very high, and the enzyme can’t be reused。 Therefore, to produce isoquercitrin more effective, a green, low cost, environmentally friendly new biological response patterns to improve manufacturing efficiency need to be investigated。 

(1) The method of naringinase-catalyzed selective hydrolysis of rutin was established。 Several factors including the substrate concentration, enzyme concentration, temperature and pH were investigated to study these effects on isoquercitrin yield。 Isoquercitrin yield was obtained to 86。64% with the conditions of pH 7 citric acid-disodium hydrogen phosphate buffer solution, 2 g/L rutin solution, 20 g/L naringinase solution in 50 °C for 1 h。

(2) Preparation of the immobilized naringinase by nanoparticles, and optimization of the preparation process。 And the ratio of the carrier and enzyme, temperature and time were studied。 The immobilization efficiency was obtained to 28。37% when the naringinase concentration was 15 g/L, graphene:enzyme solution was 4:1 (m/v) in 30 °C for 1 h。

(3) The process of enzymatic synthesis isoquercitrin by hydrolysis rutin was optimized。 In the common reactor, isoquercitrin yield was obtained to 86。64% with the optimal conditions of 2。0 g/L rutin solution, 20 g/L naringinase concentration in 50 °C for 1 h。

(4) The program of enzymatic synthesis isoquercitrin by hydrolysis rutin in microchannel reactor was optimized。 Sodium dodecyl benzene sulfonate was selected as the surface active agent。 Isoquercitrin yield was obtained to 92。78% with the conditions of rutin concentration (dissolved in pH 7 buffer solution) 0。05 g/L, temperature 55 °C, flow rate 4 μL/min。

Keywords: Nanoparticles; Immobilized enzyme; rutin; isoquercitrin; Micro reactor