嗜冷枯草芽孢杆菌低温脂肪酶纯化与酶学性质研究  

Purification and Enzymatic Characters of Low-temperature Lipase from Bacillus phychrophilus

作　　者：胡泊[1] 吴胜[2] 杨柳[2] 郑国钧[1] 孙万儒[2] 

机构地区：[1]北京化工大学生命科学与技术学院,北京100029 [2]中国科学院微生物研究所微生物资源国家重点实验室,北京100101

出　　处：《微生物学通报》2007年第3期524-527,共4页Microbiology China

基　　金：国家973项目(No.2004CB719606);微生物资源国家重点实验室开放项目(No.041014)

摘　　要：从筛选出的产低温脂肪酶的菌株发酵液中,经硫铵沉淀、疏水色谱和阴离子交换色谱纯化得到电泳纯酶。酶的最适作用温度为25℃,0℃以下仍可保持25%左右的相对酶活;在pH5.8～8.8的范围内有较高活力,其最适作用pH为7.8;对热很敏感,在60℃保温30min活性即全部丧失,具有典型的低温脂肪酶特征;酶催化不需要金属离子的参与,结构中可能含有二硫键。在25℃,pH8.0测得酶水解反应的Km值为2.65×10-5mol/L,Vmax值为5.21mmol/(L.min)。This study involves purification of a wild low-temperature lipase from a strain of Bacillus phychrophilus and analysis of its enzymatic characters. Electrophoretic pure enzyme on the SDS-PAGE was obtained through ammonium sulfate precipitation, phenyl superpose hydrophobic chromatography and Q FF anion-exchange chromatography. The optimum temperature of the enzyme is 25℃, which still has 25% relative lipase activity at 0℃. The activity of the lipase almost completely lost after incubation at 60℃ for 30 min. Lipase activity is independent of divalent cation. And the structure of this lipase may contain disulfide bond. The Km and Vmax of lipase under pH8.0 and 25℃ were 2.65 × 10^-3 mol/L and 5.21mmol/（L·min） respectively.

关 键 词：低温脂肪酶 纯化 酶学性质 

分 类 号：Q814[生物学—生物工程]