解脂耶氏酵母脂肪酶Lip2的固定化工艺及其富集鱼油DHA的分析-生物化工专业论文.docxVIP

华中科技大学硕士学位论文 华 中 科 技 大 学 硕 士 学 位 论 文 II II 16 h，可获最高 DHA 含量 32%。经过 10 次循环利用，该固定化酶仍具有初始酶活 93%的催化效率。这表明固定化 Yl Lip2 具有良好的操作稳定性。 关键词：解脂耶氏酵母脂肪酶 Lip2（Yl Lip2） 离子交换树脂 固定化 水解反应 DHA 富集 III III Abstract Yarrowia lipolytica Lip2 is one of the ideal catalysts for the enrichment of DHA in Fish Oil. However, there are some shortcomings of free Yl Lip2, such as poor operational stability, expensive price and inability to reuse and so on. These have been enormously hampered the wide application Yl Lip2. Hence，these disadvanteges must be overcame and now, enzyme immobilization technology is the mainstream in this field. This research systematically investigated the immobilization process of Yl Lip2, using macroporous resin as carrier. In addition, the application of immobilized lipase in the enrichment of docosahexenoic acid was still researched. The main work is as following: Through primarily screening on eight kinds of ion exchange resins including D311、D301R、D418、D152H、D380、D151H、D401 and D113 and four kinds macroporous resin including D3520、D4020、AB-8. D152H was choosed as the best supporter . Yl Lip2 was immobilized on the cation-exchange resin D152H by strong-acid cation exchange resin adsorption. Factors as loading amount of the lipase, immobilized time, immobilized temperature, pH of immobilized medium and drying metholds were optimized in detail. The optimum levels for the factors were: lipase loading 0.02 g per gram carrier, adsorption for 1 h, temperature 20℃, pH 6.6 of citric acid-citric acid sodium buffer, Vaccuum drying. The immobilization efficiency of the optimized immobilized enzyme was 88.17%. The specific activity of the immobilized protein was 764722 U/g with its activity recovery 177.66%. Based on single factorial experiments, the Box-Behnken method was applied to further optimize the preparation conditions. Three factors: lipase loading、buffer pH and reaction temperature were selected as the key factors affecting the specific activity of the immobilized prote