CAJ | 학술논문

在15 L的生物反应器内,对重组人胸腺素原α（huPTMAα）在大肠杆菌TG-1中经IPTG 诱导后的表达条件进行优化；应用NBS-MICROS 15 L.T.DR型生物反应器(15 L)，用工程菌E.coli pGEX-IN/PTMA,研究了诱导时不同的pH、温度、搅拌速率和IPTG浓度，利用正交试验对发酵条件进行了优化。经1次预试验和9次正式试验结果表明，huPTMAα的表达受溶氧条件的影响较显著，其中搅拌速率和通气量为最大的影响因子，当其为250r/min时，表达水平最高。经SDS-PAGE和凝胶扫描发现表达蛋白占菌体蛋白的60%以上。该实验为工程菌的中试提供了最佳的表达诱导条件，对其稳定性，高效表达具有重要的指导意义。
재15 L적생물반응기내,대중조인흉선소원α（huPTMAα）재대장간균TG-1중경IPTG 유도후적표체조건진행우화；응용NBS-MICROS 15 L.T.DR형생물반응기(15 L)，용공정균E.coli pGEX-IN/PTMA,연구료유도시불동적pH、온도、교반속솔화IPTG농도，이용정교시험대발효조건진행료우화。경1차예시험화9차정식시험결과표명，huPTMAα적표체수용양조건적영향교현저，기중교반속솔화통기량위최대적영향인자，당기위250r/min시，표체수평최고。경SDS-PAGE화응효소묘발현표체단백점균체단백적60%이상。해실험위공정균적중시제공료최가적표체유도조건，대기은정성，고효표체구유중요적지도의의。
To optimize the expression of recomb inant human prothymosinα(huPTMAα) in TG-1 induced by IPTG in 15L NBS fermento r. The E.Coli pGEX-IN/PTMA was constructed by our laboratory and differ ent pH，temperature，agitation rate and concentration of IPTG were studied by orthogonal experimental design.We found that the expression level is remarkable affected by the amount of dissolv ed oxygen, which suggests the agitation rate is the most important parameter dur ing fermentation.Examined by SDS-PAGE and gel scanning, the expression level of total protein is over 60% when agitation rate is 250r/min and therefore we ha ve established a method for high-level expression of huPTMAα on pilot-scale.