猪胰脂肪酶提取物中所含脂肪酶作为对映选择性生物催化剂的不同性质。

Different properties of the lipases contained in porcine pancreatic lipase extracts as enantioselective biocatalysts.

作者信息

Segura Rosa L, Palomo Jose M, Mateo Cesar, Cortes A, Terreni M, Fernández-Lafuente Roberto, Guisan Jose M

机构信息

Departamento de Biocatalisis, Instituto de Catalisis, CSIC, Campus UAM, 28049 Madrid, Spain.

出版信息

Biotechnol Prog. 2004 May-Jun;20(3):825-9. doi: 10.1021/bp034363p.

DOI:10.1021/bp034363p

PMID:15176888

Abstract

The porcine pancreatic lipase (PPL) extracts contain a mixture of several lipases. Their fractioning was performed by sequential adsorption via interfacial activation on supports with different hydrophobicity. A protein of 25 KDa was preferentially adsorbed on octyl-Sepharose, another protein of 33 kDa was mainly adsorbed on octadecyl-Sepabeads support, and the PPL was mainly adsorbed on the support bearing phenyl groups. The different immobilized preparations showed different properties and different response due to change in the experimental conditions. Thus, in the hydrolysis of (+/-)-2-hydroxy-4-phenylbutyric acid ethyl ester [(+/-)-1] to produce the corresponding acid [2], the octyl-25KDa preparation showed the best enantioselectivity (E) value (E = 7) at pH 5 and 25 degrees C, whereas the phenyl-PPL was the most enantioselective (E = 10) at pH 5, 4 degrees C, and 10% dioxane. Using different preparations at different pHs it was possible to resolve (+/-)-2-O-butyryl-2-phenylacetic acid [(+/-)-3] with a high E value (E > 100); for example, with octadecyl-33 KDa enzyme at pH 8.

摘要

猪胰脂肪酶（PPL）提取物包含几种脂肪酶的混合物。通过在具有不同疏水性的载体上进行界面活化的顺序吸附来对它们进行分级分离。一种25 kDa的蛋白质优先吸附在辛基琼脂糖上，另一种33 kDa的蛋白质主要吸附在十八烷基Sepabeads载体上，而PPL主要吸附在带有苯基的载体上。由于实验条件的变化，不同的固定化制剂表现出不同的性质和不同的响应。因此，在水解（±）-2-羟基-4-苯基丁酸乙酯[（±）-1]以生成相应的酸[2]时，辛基-25 kDa制剂在pH 5和25℃下显示出最佳的对映选择性（E）值（E = 7），而苯基-PPL在pH 5、4℃和10%二氧六环时对映选择性最高（E = 10）。在不同pH下使用不同的制剂，可以以高E值（E > 100）拆分（±）-2-O-丁酰基-2-苯基乙酸[（±）-3]；例如，在pH 8下使用十八烷基-33 kDa酶。