Study on immobilization of lipase B from Candida Antarctica and its application

摘要 南极假丝酵母脂肪酶B（CandidaantarcticalipaseB，CALB）是工业上广泛应用的脂肪酶。维生素A棕榈酸酯是具有良好市场前景的维生素A衍生物。利用南极假丝酵母脂肪酶B催化合成维生素A棕榈酸酯具有条件温和，反应选择性高，副反应少，产品的稳定性好，绿色环保等优点。 本文首先培养重组毕赤酵母获得高浓度CALB的发酵上清液。发酵144h后，游离酶酶活可达108U/mL，目标蛋白占总蛋白含量的百分比为63.4%。通过对游离酶固定化方法的比较，发现与共价法相比，吸附法操作简单，同时对酶活破坏小。采用大孔吸附树脂HPD826吸附固定脂肪酶，其催化月桂酸酯化的活性可达9200PLU...Abstract Candida antarctica lipase B (CALB) has been widely used in industry and vitamin A palmitate has great market prospects. The synthesis of vitamin A palmitate catalyzed by Candida antarctica lipase B has many advantages including mild conditions, high reaction selectivity, less side effects, good product stability, environmental friendly etc. Firstly, the high concentration of CALB were o...学位：工学硕士院系专业：化学化工学院_化学工程学号：2062013115148

Preparation of fotemustine-loaded polybutylcyanoacrylate nanoparticles

目的优化工艺制备福莫司汀聚氰基丙烯酸正丁酯纳米粒(fCnu-PbCA-nP)。方法以α-氰基丙烯酸正丁酯(bCA)为载体,采用乳化聚合法制备fCnu-PbCA-nP,并加以聚乙二醇20000(PEg20000)进行表面修饰,通过考察粒径和包封率两个指标,在单因素实验初选的基础上,正交设计法优化处方和制备工艺。结果制备fCnu-PbCA-nP的优化条件为bCA单体体积分数0.8%(V/V)、fCnu 20 Mg、PEg20000浓度2.0%,按优化条件所制备的fCnu-PbCA-nP的粒径为(124.6±5.2)nM,多分散系数(PdI)范围为0.07--0.16,包封率(64.12±2.36)%,载药量(7.28±0.76)%。结论通过优化处方和制备工艺,采用乳化聚合法可制备出fCnu-PbCA-nP,对拓展fCnu临床给药新剂型提供一定的参考。AIM To prepare fotemustine polybutylcyanoacrylate nanoparticles(FCNU-PBCA-NP) with optimized process.METHODS FCNU-PBCA-NP was prepared by emulsion polymerization with the α-butylcyanoacrylate(BCA) as its carrier and the surface of the nanoparticles was modified with polyethylene glycol 20000(PEG20000).Single factor test and orthogonal design were carried out to optimize the preparing technology according to the particle size and the entrapment efficiency of FCNU-PBCA-NP.RESULTS The optimal conditions for the preparation of FCNU-PBCA-NP were 0.8% BCA monomer(V/V),20 mg fotemustine and 2.0% PEG20000(m/V).On the basis of the above conditions,the mean particle size of the NP was(124.6±5.2)nm and the polydispersity index(PDI) was 0.07-0.16,the average entrapment efficiency and drug loading was(64.12±2.36)% and(7.28±0.76)%,respectively.CONCLUSION An optimized nanoparticle drug delivery system is obtained by emulsion polymerization and provides a new direction for fotemutine dosage forms in future.福建省自然科学基金(2006J0188);厦门市科技局基金(3502Z20064013

Synthesis of Vitamin a Palmitate Catalyzed by Lipase in Fixed Bed Reactor

探讨了利用固定床反应器进行脂肪酶催化合成维生素A棕榈酸酯的可行性．利用HPD826吸附法获得固定化酶活力达到9200U／g，确定了反应介质为石油醚，进料流量8mL／min，反应温度为35℃，维生素A乙酯及棕榈酸的浓度分别为3．6g／L和14g／L．在此优化条件下反应，转化率可达80％，维生素A棕榈酸酯的生产能力为281g／（L·h）．反应器连续运行12h，转化率维持在68％以上．结果表明，利用固定床反应器进行酶促合成维生素A棕榈酸酯可行，可进一步深入研究．The feasibility of using fixed bed reactor for synthesis of vitamin A palmitate catalyzed by li- pase was investigated in this work.First, the lipase was immobilized by adsorption with HPD826 as car- rier and give the activity of 9200 U/g.Then the appropriate reaction conditions were determined：the re- action media is petroleum ether,the feed flow rate is 8 mL/min,the temperature is 35 ℃ ,the concen- trations of vitamin A acetate and palmitic acid are 3.6 and 14 g/L.Under these conditions,the conver- sion and productivity of vitamin A palmitate can reach 80% and 281 g/（L · h） ,respectively.Finally,the reactor was continuously run for 12 h,the conversion held upon 68%.These results showed that the synthesis of vitamin A palmitate by immobilized lipase in fixed bed reactor was feasible and worth fur- ther investigation.厦门市科技计划项目（3502220143008