约氏不动杆菌 LP28低温碱性脂肪酶分离纯化及洗涤性能分析

摘要

对菌株LP28发酵产的低温碱性脂肪酶进行了分离纯化，菌体发酵液经过离心、硫酸铵沉淀、DEAE-Sepha-rose Fast Flow离子交换及Sephadex G-75凝胶过滤等步骤处理，比活力达76．66 U／mg，纯化倍数达36．16倍，回收率为14％。菌株LP28所产的脂肪酶洗涤性能分析显示，洗涤温度在30℃、pH 8．0、加酶量为100 U、洗涤剂添加量为0．25％时，其洗涤效果最佳，去油率达35％。且该酶对多种底物具有特异性，并对蛋白酶及市售洗衣粉也具有一定的稳定性。说明Acinetobacter johnsonii LP28脂肪酶具有洗涤剂添加剂应用前景。%A low temperature alkaline lipase from Acinetobacter johnsonii LP28 was isolated and purified u-sing ammonium sulfate precipitation ,Fast Flow DEAE-Sepharose ion exchange and G-75 Sephadex gel fil-tration.The specific activity was 76.66 U/mg,and this purification protocol resulted in a 36.16 fold puri-fication of lipase with 14%final yield.A study on the washing performance of the lipase shows that its op-timum temperature was 30℃and pH was 8.0.The optimal enzyme dosage and detergent dosage were 100 U and 0 .25%respectively .Under the optimum conditions , the detergency was 35%.Results shows that the lipase is specific to a variety of substrates and has a certain stability against protease and commercial washing powder ,which has good potential on detergent and additive .