培养基优化设计提升多拉菌素生物发酵水平  

Improving the biosynthesis of doramectin production by mutant strain Streptomyces avermitilis 3D-5 by optimizing fermentation medium

作　　者：赵明霞 栗波 蔡子豪 薛佳韵 王泽建[3] 梁剑光[1,4] Zhao Ming-xia;Li Bo;Cai Zi-hao;Xue Jia-yun;Wang Ze-jian;Liang Jian-guang(School of Biological and Food Engneering,Changshu Institute of Technology,Changshu 215500;College of Pharmaceutical Science,Soochow University,Suzhou 215123;State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai Institute of Biomanufacturing Technology&Collaborative Innovation Center,Shanghai 200237;College of Pharmaceutical and Life Sciences,Changzhou University,Changzhou 213164)

机构地区：[1]常熟理工学院生物与食品工程学院,常熟215500 [2]苏州大学医学部药学院,苏州215123 [3]华东理工大学生物反应器工程国家重点实验室,上海200237 [4]常州大学制药与生命科学学院,常州213164

出　　处：《中国抗生素杂志》2020年第11期1121-1132,共12页Chinese Journal of Antibiotics

基　　金：国家重点研发计划项目(No.2017ZX07402003)。

摘　　要：运用响应面法优化了多拉菌素生产菌的发酵培养基。首先通过单因素实验发现正效应因子;接着采用Plackett-Burman(P-B)设计确定了黄豆饼粉、麦芽糊精和MgSO4是影响多拉菌素产量的显著因素;然后利用最陡爬坡试验分别找到3个因素的合理浓度范围;并进一步利用中心组合设计优化了黄豆饼粉、麦芽糊精和MgSO4的最佳浓度配比。筛选并优化得到了最适的培养基浓度为黄豆饼粉17.30g/L,麦芽糊精77.30g/L,MgSO4 1.50g/L,基于此,效价达到了589.43mg/L,验证实验结果与模型预测基本吻合,优化后的培养基工艺能够提升多拉菌素发酵单位20.37%。5L反应罐上发酵过程生理代谢参数变化表明:优化的培养基能够加促菌体的比生长速率,维持较高的氧消耗速率和产物合成速率,大幅度提升了多拉菌素的发酵生产效率。Objective The fermentation medium compositions for doramectin production by mutant strain Streptomyces avermitilis 3D-5 was optimized through response surface methodology(RSM). Methods Firstly, positive effect factors on doramectin production were found by the one-factor-at-a-time method, and then, soybean cake powder, maltodextrin and MgSO4 were screened as the significant factors for doramectin biosynthesis with the PlackettBurman(P-B) design. Furthermore, the steepest ascent path was applied to investigate the optimal range of the medium composition. Finally, the optimal combination ratio of soybean cake powder, maltodextrin and MgSO4 was determined by the Central composite design(CCD). Results Results showed that the doramectin yield rose up to 589.43 mg/L, under the achieved compositions of soybean cake powder 17.30 g/L, maltodextrin 77.30 g/L, and MgSO4 1.50 g/L. The verification experiment showed the doramectin yield was consistent with the model prediction(589.43 mg/L), which was 20.37% higher than that of the original medium condition. Conclusion The fermentation process physiological parameters obtained for the strain S. avermitilis 3D-5 in the 5 L bioreactor showed that the specific growth rate was significantly enhanced under the optimized medium, and the higher oxygen uptake rate accompanied with higher specific production rate. The specific dormadetin production rate was markedly enhanced in the optimized medium.

关 键 词：响应面 多拉菌素 培养基优化 发酵 

分 类 号：R978.1[医药卫生—药品]