Baillargeon M W, McCarthy S G
U.S. Department of Agriculture, Eastern Regional Research Center, Philadelphia, Pennsylvania 19118.
Lipids. 1991 Oct;26(10):831-6. doi: 10.1007/BF02536165.
Lipases from Geotrichum candidum NRRL Y-553 are of interest because of their unique specificity for cis-9-unsaturated fatty acids relative to both stearic and palmitic acids. The lipases were partially purified by chromatography on Octyl Sepharose, AG MP-1 macroporous anion exchanger, and chromatofocusing resin. The preparation was found to contain multiple, glycosylated lipases varying slightly in pI (pI 4.88, 4.78, 4.65, 4.57 and 4.52) as judged by both activity and silver staining. The molecular mass determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis was 64 kilodaltons for the main species, with minor species of 60 and 57 kilodaltons present as well. The specificity of the crude lipases for hydrolysis of 4-methylumbelliferyl esters of oleic vs. palmitic acid was 20-to-1. The specificity of the purified, partially separated lipases was similar to that of the crude preparation. Thus the lipases could be used even in crude form for the hydrolysis and restructuring of triacylglycerols on a large scale.
白地霉NRRL Y-553产生的脂肪酶备受关注，因为相对于硬脂酸和棕榈酸，它们对顺式-9-不饱和脂肪酸具有独特的特异性。通过在辛基琼脂糖、AG MP-1大孔阴离子交换剂和色谱聚焦树脂上进行色谱分离，对脂肪酶进行了部分纯化。通过活性和银染判断，发现该制剂含有多种糖基化脂肪酶，其pI略有不同（pI 4.88、4.78、4.65、4.57和4.52）。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳测定，主要种类的分子量为64千道尔顿，同时还存在60和57千道尔顿的次要种类。粗脂肪酶对油酸和棕榈酸的4-甲基伞形酮酯的水解特异性为20比1。纯化的、部分分离的脂肪酶的特异性与粗制剂相似。因此，这些脂肪酶即使以粗形式也可用于大规模水解和重构三酰甘油。
