Baillargeon M W
Agricultural Research Service, Eastern Regional Research Center, U.S. Department of Agricuture, 600 East Mermaid Lane, 19118, Philadelphia, PA.
Lipids. 1990 Dec;25(12):841-8. doi: 10.1007/BF02535907.
A crude, commercialGeotrichum candidum lipase (EC 3.1.1.3) preparation (Amano GC-20) was purified by hydrophobic interaction chromatography on Octyl Sepharose. The purified enzyme is a microheterogeneous glycoprotein containing isozymes varying in molecular weight, pI and specificity. It consists of 64, 62 and 59 kDa species as determined by denaturing polyacrylamide gel electrophoresis. Five isozymes (pI 4.40, 4.47, 4.58, 4.67 and 4.72) are detected by isoelectric focusing using both silver and activity stains. Chromatofocusing was used to separate the isozymes according to pI. Although all the isozymes are specific for oleatevs stearate esters, one isozyme (pI 4.72) is also specific for oleatevs palmitate. The number of isozymes is reduced to two (pI 4.67 and 4.72) after carbohydrate removal using endoglycosidase F/N-glycosidase. These isozymes may be products of two lipase genes.
一种粗制的商品白地霉脂肪酶（EC 3.1.1.3）制剂（天野GC - 20）通过在辛基琼脂糖上进行疏水相互作用色谱法进行纯化。纯化后的酶是一种微不均一的糖蛋白，含有分子量、等电点和特异性不同的同工酶。通过变性聚丙烯酰胺凝胶电泳测定，它由64 kDa、62 kDa和59 kDa的组分组成。使用银染和活性染色通过等电聚焦检测到五种同工酶（等电点4.40、4.47、4.58、4.67和4.72）。使用色谱聚焦根据等电点分离同工酶。尽管所有同工酶对油酸酯与硬脂酸酯具有特异性，但一种同工酶（等电点4.72）对油酸酯与棕榈酸酯也具有特异性。使用内切糖苷酶F/N - 糖苷酶去除碳水化合物后，同工酶数量减少到两种（等电点4.67和4.72）。这些同工酶可能是两个脂肪酶基因的产物。
