嗜热果糖苷酶的基因克隆及表达优化  

Cloning and optimization expression for thermophilic fructosidase

作　　者：毕云枫[1] 徐琳琳[1] 姜珊[1] 李娜[1] 王溪竹[1] 王玉华[1] 

机构地区：[1]吉林农业大学食品科学与工程学院,长春130118

出　　处：《中国食品添加剂》2017年第4期74-82,共9页China Food Additives

基　　金：吉林省教育厅"十二五"科学技术研究项目(吉教科合字[2015]第207号);吉林省科技厅科技创新人才培育计划项目(项目编号:20140519011JH)

摘　　要：从嗜热菌Thermotoga neapolitana DSM4359得到嗜热果糖苷酶基因片段,以p ET-28a(+)为表达载体,在Escherichia.coil BL21(DE3)中高效表达。由SDS-PAGE检测可得构建后嗜热果糖苷酶的分子量约为60 KDa。通过单因素试验和响应面法对该嗜热果糖苷酶的产酶培养基进行优化选择,最终确定最佳产酶培养基为:蔗糖10.11 g/L,硝酸铵12.30 g/L,安琪酵母浸粉FM905 8.24 g/L,MgSO_4·7H_2O 5.61 mmol/L,NaCl 10 g/L。相比优化前,经优化后的培养基所产嗜热果糖苷酶酶活提高了近2.1倍。达到了提高嗜热果糖苷酶表达量的目的。Genes of fructosidase was extracted from thermophilic Thermotoga neapolitana DSM4359,pET-28a（＋）expression vector was used and overexpressed in Escherichia.coil BL21（DE3）.SDS-PAGE shows the molecular weight of the thermophilic fructosidase is 60 KDa.In order to increase thermophilic fructosidase expression volume,the expression conditions were optimized.By single factor tests and response surface method the best medium was：sucrose 10.11 g/L,ammonium nitrate 12.30 g/L,angel yeast extract FM905 8.24 g/L,MgSO4· 7H2O 5.61 mmol/L,NaCl 10 g/L.The optimized medium can increase thermophilic fructosidase activity to 2.1 times.The results meet the purpose of improving the expression of thermophilic fructosidase.

关 键 词：嗜热果糖苷酶 基因克隆 表达 优化 

分 类 号：TS201.3[轻工技术与工程—食品科学]