交联酶聚集体与仿生硅化技术结合制备固定化脂肪酶

Preparation of Immobilized Lipase through Combination of Cross-Linked Enzyme Aggregates and Biomimetic Silicification

将交联酶聚集体(CLEAs)与仿生硅化技术相结合,制备了交联脂肪酶Candidasp.99-125杂化生物催化剂.以京尼平为交联剂,在最佳条件下制得的脂肪酶CLEAs的酶活达771U/g,回收率达75%;保护剂聚乙烯亚胺(PEI)与Candidasp.99-125脂肪酶共沉淀制备P/CLEAs,其酶活达897U/g,回收率约88%;利用PEI的诱导作用,在P/CLEAs表面形成氧化硅涂层,制得的脂肪酶CLEAs(Coated-CLEAs)显示出良好的稳定性,特别是其抗蛋白酶水解能力、有机溶剂耐受能力、重复使用性能等方面明显提高.

The hybrid biocatalyst was prepared by the combination of cross-linked enzyme aggregates（CLEAs） and biomimetic silicification,where the lipase Candida sp.99-125 was used as model enzyme.The preparation conditions of lipase-CLEAs were optimized and the resulting activity of 771 U/g was obtained.Under the optimum conditions,75% of the activity recovery was obtained.P/CLEAs was prepared by coprecipitation of lipase with polyethylene-imine（PEI）,and the resulting activity was 897 U/g.The activity recovery of P/CLEAs was about 88%.The silica coating was formed on the outside surface of P/CLEAs via the induction of PEI.The resulting Coated-CLEAs showed high reusability.The stability of the Coated-CLEAs,including the resistance to trypsin and organic solvents,was also significantly improved.