CAJ | 학술논문

为了充分发挥出突变株的高转化活性及其优良特性,采用响应面试验设计方法优化(R)-肾上腺素高产菌株Kocuria rhizophila(K.rhizophila).H5401的发酵培养基.用Plackett-Burman设计法研究初始发酵培养基中各成分对(R)-肾上腺素产率的影响,筛选出有显著效应的3个因素:甘油、蛋白胨和NH4H2PO4.利用最陡爬坡法逼近最大响应区域.通过Box-Behnken中心组合实验和响应面分析方法确定出3个主要影响因素的最佳浓度.实验结果经过回归分析拟合出1个2次方程,用其预报的最优培养基做摇瓶发酵试验,(R)-肾上腺素产率达61.07%,比优化前提高了24.89%,表明响应面试验设计优化方法有效提高了(R)-肾上腺素的产量,降低了生产成本,为后续放大实验提供依据.
위료충분발휘출돌변주적고전화활성급기우량특성,채용향응면시험설계방법우화(R)-신상선소고산균주Kocuria rhizophila(K.rhizophila).H5401적발효배양기.용Plackett-Burman설계법연구초시발효배양기중각성분대(R)-신상선소산솔적영향,사선출유현저효응적3개인소:감유、단백동화NH4H2PO4.이용최두파파법핍근최대향응구역.통과Box-Behnken중심조합실험화향응면분석방법학정출3개주요영향인소적최가농도.실험결과경과회귀분석의합출1개2차방정,용기예보적최우배양기주요병발효시험,(R)-신상선소산솔체61.07%,비우화전제고료24.89%,표명향응면시험설계우화방법유효제고료(R)-신상선소적산량,강저료생산성본,위후속방대실험제공의거.
Aiming at increasing the rate of production, the fermentation medium of Kocuria rhizophila(K. rhizophila). H5401 highly producing (R)-adrenaline was optimized by using response surface analyzing method(RSAM). In the first step of optimization, the im-pact of initial medium constituents on (R)-adrenaline production was studied by Plackett-Burman design. Glycerol, Peptone and NH4H2PO4 were found to be the important factors affecting (R) -adrenaline production significantly. In our work, the greatest response area was estimated by the design of steepest ascent approach. Box-Behnken central composite design and RSAM were applied to deter-mine the optimal concentration of each significant variable. The quadratic model established by regression analysis facilitated the pre-diction of the optimum medium composition for the fermentation process, and the results of the experiment were in agreement with our proposed model. (R)-adrenaline yield with ultimate value 61.07% was observed after the optimization carried out according to RSAM design, which was increased by 24. 89% in the original medium. The optimized condition allowed (R)-adrenaline production to in-crease effectively.