伯克霍尔德菌ZYB002胞外脂肪酶的分离纯化及其酶学性质分析  

Purification and Enzymatic Characterization of the Extracellular Lipase from Burkholderia sp.ZYB002

作　　者：刘艳如[1] 邱芳锦 舒正玉[1] 王作镇 邱黎清[1] 李欣[1] 江贤章[1] 黄建忠[1] 

机构地区：[1]福建师范大学工业微生物发酵技术国家地方联合工程研究中心,福建师范大学教育部工业微生物工程中心,福建师范大学生命科学学院,福建福州350117

出　　处：《福建师范大学学报（自然科学版）》2014年第3期100-105,共6页Journal of Fujian Normal University：Natural Science Edition

基　　金：国家自然科学基金资助项目(31370802);福建省科技厅重点项目(2013H0021);福建省自然科学基金杰青项目(2009J06013)

摘　　要：伯克霍尔德菌ZYB002胞外脂肪酶发酵上清液经冻干、透析、DEAESepharose Fast Flow层析柱和SephadexG-75层析柱等步骤纯化后，获得电泳纯的脂肪酶．纯化后的脂肪酶分子质量为33ku，水解棕榈酸对硝基苯的比活力为1902．5U·mg^-1．脂肪酶催化水解反应的最适温度和最适pH值分别为50℃和8．5；在30-65℃和pH3．0-10．0的范围内保持相对稳定．脂肪酶对烷烃、非离子型表面活性剂等有机溶剂具有较好的耐受性．在最适条件下，脂肪酶水解pNPP的k和‰。。分别为3．79mmol·L^-1和714．29μmol·mg^-1·min^-1．The extracellular lipase from Burkholderia sp. ZYB002 was purified to homogeneity by the following procedures： lyophilization, dialysis, DEAE Sepharose Fast Flow anion-exchange chromatography and Sephadex G-75 gel-filtration chromatography, respectively. The relative molec- ular mass of the purified lipase was 33 ku and the specific activity for pNPP hydrolysis was 1 902.5 U·mg-1 The optimal temperature and pH for hydrolysis reaction of the purified lipase were 50 ℃ and 8.5, respectively. The lipase displayed excellent resistance to heat and basic solution in the range of 30 to 65 ℃ and pH 3.0 to 10. 0, respectively. The lipase also exhibited excellent tolerance to several organic solvents, including alkane, nonionic surfactants. Under the optimal hydrolysis conditions, the Km and VmaxOf the purified lipase calculated from the Lineweaver-Burk plot using p- nitrophenyl palmitate as hydrolysis substrate were 3.79 mmol·L-1 and 714. 29 μmol·mg^-1·min^-1, respectively.

关 键 词：伯克霍尔德菌ZYB002 胞外脂肪酶 分离纯化 酶学性质 

分 类 号：Q556[生物学—生物化学]