响应面法优化脂肪酶的固定化条件及其水解橄榄油的特性研究  

Optimization of Immobilization of Lipase Using Response Surface Methodology and Its Performances in Olive Oil Hydrolysis

作　　者：刘自琴[1] 黄惠华[1] 

机构地区：[1]华南理工大学轻工与食品学院,广东广州510640

出　　处：《食品科学》2012年第3期184-189,共6页Food Science

基　　金：国家"863"计划项目(2010AA101505)

摘　　要：采用Sepharose CL-4B和7种大孔树脂为载体固定Palatase 20000L脂肪酶,对载体进行筛选,以酶活力为指标,采用响应面法优化固定化条件,并考察所制得固定化酶的稳定性和水解橄榄油的酶学性质。结果表明:以大孔树脂HPD-600为载体制得的固定化酶具有较高的活性和良好的稳定性。其最优的固定化条件为:pH3.9,酶与载体比例为9.1mg/g,吸附时间1.8h。在最优条件下制得固定化酶在最适合条件下测得的酶活力达到1440U/g,酶活回收率大于50%。固定化酶最适作用温度为50℃,最适作用pH值为8.0。固定化酶的Km值为0.130g/mL,高于游离酶的Km(0.069g/mL)。固定化酶的热稳定性有一定程度的提高,其重复操作5次后相对酶活力仍保持在58%以上。Different carriers such as Sepharose CL-4B and seven kinds of macroporous resins were used to immobilize lipase Palatase 20000L. The lipase showed higher activity and better stablity when immpbilized on macroporous resin type HPD-600 compared with other carriers. Ressponse surface methodolgy was employed to optimize key conditions for the immobilization of lipase Palatase 20000L on macroporous resin type HPD-600 to achieve maximum immobilized lipase activity. The optimal immobilization conditions were found as follows： pH 3.9, enzyme-to-carrier ratio 9.1 mg/g, and adsorption time 1.8 h. Under these conditions, the immobilized lipase achieved its maximum activity, 1440 U/g and a rantivity ecovery above 50%. The optimal reacton temperature of the immobilized lipase was 50 ℃, the optimal reaction pH 8.0, and the Km value 0.130 g/mL, which was higher than that （0.069 g/mL） of the free form. Moreover, the immobilized lipase showed higher thermal stability and maitained over 58% of its original activity after the fifth repeated use.

关 键 词：Palatase20000L脂肪酶 固定化 稳定性 响应面法 

分 类 号：Q814.2[生物学—生物工程]