重组大肠杆菌右旋糖酐蔗糖酶的表达条件优化  

Optimization of culture conditions for recombinant dextransucrase expression

作　　者：张洪斌[1] 胡雪芹[1] 冒小青[1] 王雅洁[1] 

机构地区：[1]合肥工业大学化工学院制药工程系,合肥230009

出　　处：《生物工程学报》2009年第12期2022-2028,共7页Chinese Journal of Biotechnology

基　　金：安徽高校省级自然科学研究重点项目(No.KJ2008A067);合肥工业大学博士基金(No.GDBJ2008-021)资助~~

摘　　要：通过设计正交实验,考察了培养基中各组分及其浓度对右旋糖酐蔗糖酶工程菌Escherichia coli BL21(DE3)/pET28-dexYG诱导产酶结果的影响。在获得最佳培养条件的基础上,考察温度、蔗糖浓度和pH值对右旋糖酐产量的影响。结果表明:菌浓OD600达到2.0时,加入异丙基硫代-β-D-呋喃半乳糖苷(IPTG)至0.25mmol/L,25°C诱导培养4h,产酶活力最高,达到110.16U/mL,蔗糖浓度对产量的影响比较显著。研究结果得到高效表达的培养条件,为实现该酶的工业化应用打下了基础。We optimized the medium for recombinant dextransucrase expression in engineering strain Escherichia coli BL21 （DE3）/pET28-dexYG by an Orthogonal experiment.After the medium had been decided,we studied the effect of temperature,sucrose concentration and pH value on the yield.The results indicated that optimal conditions were adding IPTG of 0.25 mmol/L when OD600 reached 2.0 and cultivation lasted for 4 h at 25°C.Under the selected medium and these conditions,the dextransucrase activity expressed by the engineering strain was high activity the dextran yield grately. The results for dextransucrase dextransucrase. Maximal activity reached 110.16 U/mL sucrose concentration effects expression would provide foundation for industrial application of

关 键 词：右旋糖酐蔗糖酶 表达 优化 右旋糖酐 

分 类 号：Q78[生物学—分子生物学]