CAJ | 학술논문

以本实验室前期构建的重组大肠杆菌工程菌株为基础，为提高阿魏酸酯酶的表达量，通过试验设计，对工程菌株目的蛋白可溶表达条件进行优化。实验采用250 mL三角瓶中，内含50 mL（ Amp 100 mg／L）的LB培养基，分别研究诱导温度、诱导物IPTG浓度、诱导时机和诱导时间等对蛋白可溶表达量的影响。确定了阿魏酸酯酶工程菌可溶性表达最优条件为：过夜培养菌体以1∶100比例接种新鲜培养基，摇床转速200 rmp／min，在37℃下培养至OD600值为1．0时，添加终浓度为0．01 mM的IPTG进行诱导，诱导温度30℃，诱导时长为9 h，最终单位发酵液的酶活力为400 U／L。以上数据为阿魏酸酯酶重组工程菌的工业应用奠定了基础。
이본실험실전기구건적중조대장간균공정균주위기출，위제고아위산지매적표체량，통과시험설계，대공정균주목적단백가용표체조건진행우화。실험채용250 mL삼각병중，내함50 mL（ Amp 100 mg／L）적LB배양기，분별연구유도온도、유도물IPTG농도、유도시궤화유도시간등대단백가용표체량적영향。학정료아위산지매공정균가용성표체최우조건위：과야배양균체이1∶100비례접충신선배양기，요상전속200 rmp／min，재37℃하배양지OD600치위1．0시，첨가종농도위0．01 mM적IPTG진행유도，유도온도30℃，유도시장위9 h，최종단위발효액적매활력위400 U／L。이상수거위아위산지매중조공정균적공업응용전정료기출。
On the basis of the recombinant E.coli strains which were built in our laboratory,in order to improve the expression level of feruloyl esterase , the conditions of soluble protein expression were optimized through experimental design.Using 250 mL Erlenmeyer flask containing 50 mL (Amp 100 mg/L) medium,induction temperature,inducer concentration,induced expression of soluble protein timing and induction time were studied .Identified feruloyl esterase engineered bacteria optimal conditions for soluble expression were incubated at 37 ℃ to OD600 value of 1.0,adding a final concentration of 0.01mM of IPTG,induction temperature 30 ℃, shaking speed 200 rmp/min, inoculum of 1%, while the induction time 9 h.After the fermentation 400 U/L enzyme activity was obtained.The above data make a basis for the recombinant engineering bacteria pilot fermentation.