文献类型：期刊文献

中文题名：响应面法优化林可霉素产量与组分

英文题名：Optimization of lincomycin production and component by response surface methodology

作者：庄智慧[1];高淑红[1];何岫峰[2];杨灿灿[2];秦振[2]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]天方药业有限公司,驻马店463000

年份：2018

卷号：43

期号：8

起止页码：1049

中文期刊名：中国抗生素杂志

外文期刊名：Chinese Journal of Antibiotics

收录：CSTPCD;;Scopus;CSCD:【CSCD_E2017_2018】；

基金：国家自然科学基金青年基金(No.21506056)

语种：中文

中文关键词：林可链霉菌;林可霉素;响应面法;发酵优化

外文关键词：Streptomyces lincolnensis;Lincomycin;Response surface methodology;Fermentation and optimization

摘要：以林可霉素A(Lin A)产量提高以及林可霉素B组分(Lin B)含量降低为目标,在单因素实验基础上,利用响应面法对林可链霉菌18-8菌株的含硫前体物质(硫酸钠、甲硫氨酸、半胱氨酸)与戊糖磷酸途径(HMP)的关键前体(葡萄糖酸钙、葡萄糖酸钠)以及相关醇类、离子(肌醇、氯化钴)等物质进行组合优化。首先对7个因素进行Plackett-Burman实验,筛选得到3个显著性因子:葡萄糖酸钙、肌醇和氯化钴。再通过Box-Behnken设计3因素3水平实验,利用Design-Expert 8.5软件进行回归分析,得到最佳优化条件为:氯化钴7.97mg/L、葡萄糖酸钙6.0g/L、肌醇0.42g/L。摇瓶验证Lin A液相效价为5210mg/L,比优化前提高21%,Lin B含量为4.3%,比出发菌株的Lin B含量(6.0%)降低39.5%;在15L发酵罐中进行发酵实验,Lin A液相效价可达8980mg/L,比对照(6630mg/L)提高35%,Lin B含量在90h达到最低含量2.4%,相比对照Lin B含量(4.8%)降低50%,效果显著。
In order to optimize the fermentation process of lincomycin, the increased yield of lincomycin A and the decreased content of lincomycin B as the evaluation index and the response surface method were used to optimize the content of fermentation medium components such as sulfur precursor （sodium sulfate, methionine, and cysteine）, precursor of pentose phosphate pathway （HMP） （calcium gluconate and sodium gluconate）, and related alcohol and ions （inositol and cobalt chloride） during fermentation by Streptomyces lincolnesis 18-8. Three significant factors （calcium gluconate, inositol, and cobalt chloride） were chosen for further study by using the Plackett-Burman design experiments. The Box-Behnken design was used to conduct a three-factor three-level experiment. The Design- Expert 8.5 software was used for the regression analysis. The optimal fermentation conditions were determined as follows： 7.97mg/L cobalt chloride, 6.0g/L calcium gluconate, and 0.42g/L inositol. In the shake flask experiment, the titer of lincomycin A was 5,210rag/L, which was increased by 21% in comparison with that of the pre-optimization fermentation. The content of lincomycin B was 4.3%, which was 39.5% lower than that of the pre-optimization fermentation. In the 15L bioreactor fermentation, the titer of lincomycin A was 8,980mg/L, 35% higher than that of the control （6630mg/L）, and the content of lincomycin B was decreased to 2.4% at 90h, 50% lower than that of the control （4.8%）. This study shows promising effects using the response surface methodology for the optimization of lincomycin fermentation process.