Kakugawa Koji, Shobayashi Megumi, Suzuki Osamu, Miyakawa Tokichi
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima, Japan.
Biosci Biotechnol Biochem. 2002 May;66(5):978-85. doi: 10.1271/bbb.66.978.
An extracellular lipase produced by the glycolipid-producing yeast Kurtzmanomyces sp. I-11 was purified by ammonium sulfate precipitation and column chromatographies on DEAE-Sephadex A-25, SP-Sephadex C-50, and Sephadex G-100. Based on the analysis of the purified lipase on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the purified lipase was judged to be homogeneous and its molecular mass was estimated to be approximately 49 kDa. The optimum temperature for the activity was 75 degrees C, and the activity was very stable at temperatures below 70 degrees C. The active pH range of this lipase was 1.9-7.2, and the activity was stable at pH below 7.1. The lipase showed a preference for C18 acyl groups by measurements with p-nitrophenyl esters and triglycerides as substrates. The lipase was very stable in the presence of various organic solvents at a concentration of 40%. Although the N-terminal sequence of the Kurtzmanomyces lipase was very similar to that of lipase A from Candida antarctica, the pH profiles of the two lipases were significantly different.
由产糖脂酵母库兹酵母属I-11产生的一种胞外脂肪酶，通过硫酸铵沉淀以及在DEAE-葡聚糖A-25、SP-葡聚糖C-50和葡聚糖G-100上的柱色谱法进行纯化。基于十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对纯化脂肪酶的分析，判断纯化后的脂肪酶为均一的，其分子量估计约为49 kDa。该脂肪酶的最适温度为75℃，在70℃以下温度时活性非常稳定。此脂肪酶的活性pH范围为1.9 - 7.2，在pH低于7.1时活性稳定。以对硝基苯酯和甘油三酯为底物进行测定时，该脂肪酶对C18酰基表现出偏好。在40%浓度的各种有机溶剂存在下，该脂肪酶非常稳定。尽管库兹酵母脂肪酶的N端序列与南极假丝酵母脂肪酶A的序列非常相似，但这两种脂肪酶的pH曲线显著不同。
