交联烯丙基葡聚糖凝胶共价偶联脂肪酶

Immobilization of Lipase by Covalent Binding on Cross-linked Copolymer of Allyl Dextran and N,N'-Methylenebisacrylamide

研究了以交联烯丙基葡聚糖凝胶 (CADB)为载体 ,对 - β-硫酸酯乙砜基苯胺 (SESA)为偶联剂固定化脂肪酶的工艺条件 ,并考察了固定化脂肪酶的稳定性。试验结果表明 :CADB与 SESA反应的最适条件是 p H=1 2 .0 ,SESA与CADB质量比为 6∶ 1 ,反应生成的对氨基苯磺酰乙基交联烯丙基葡聚糖凝胶 (ABSE- CADB)经重氮化后与脂肪酶偶联的最适条件是 p H7.5,偶联时间≥ 1 2 h。加酶量为 40 0 u/g～ 1 2 0 0 u/g(载体 )时 ,所得固定化酶活力为 1 93u/g～ 448u/g(载体 ) ,此时固定化酶的活力回收率较高 (>37% )。固定化脂肪酶稳定性较高 ,其操作半衰期约为 72 0 h。

The conditions of the lipase immobilized on the cross linked copolymer of allyl dextran and N,N' Methylenebisacrylamide(CADB)with the aid of p β sulfulic acid ester ethyl sulfone aniline(SESA) was investigated.It is shown that the optimal conditions for the reaction of CADB with SESA are pH 12.0 and m(SESA)∶m(CADB)=6∶1.The product p aminobenzart sulfanily ethyl CADB(ABSE CADB)was diazotized and linked with lipase.The optimal pH for lipase linked to diao compound is 7.5.The link time is at last 12h.When the amount of lipase is 400u/(g CADB)～1200u/(g CADB),the activity of immobilized lipase is 193u/(g CADB)～448u/(g CADB)and the activity recovery is above 37%.The immobilized lipase is fairly stability,its operational half life is about 720 hours.