优化培养基组分提高猪链球菌2型NT15B菌株活菌数的研究  

Study on the Optimization of Medium Composition to Increase the Viable Counts of Streptococcus suis Type 2 NT15B Strain

作　　者：马林丽 徐国强[1,2,4] 张秋萍 车巧林 史劲松 许正宏 MA Linli;XU Guoqiang;ZHANG Qiuping;CHE Qiaolin;SHI Jinsong;XU Zhenghong(The Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Bioengineering,Jiangnan University,Wuxi 214122,China;National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing,Jiangnan University,Wuxi 214122,China;School of Life Science and Health Engineering,Jiangnan University,Wuxi 214122,China;Jiangsu Provincial Engineering Research Center for Bioactive Product Processing Technology,Jiangnan University,Wuxi 214122,China;Jiangsu Nannong Hi-Tech Co.,Ltd.,Wuxi 214405,China)

机构地区：[1]江南大学生物工程学院,工业生物技术教育部重点实验室,无锡214122 [2]江南大学,粮食发酵与食品生物制造国家工程研究中心,无锡214122 [3]江南大学生命科学与健康工程学院,无锡214122 [4]江南大学,江苏省生物活性制品加工工程技术研究中心,无锡214122 [5]江苏南农高科技股份有限公司,无锡214405

出　　处：《中国畜牧兽医》2024年第7期2776-2786,共11页China Animal Husbandry & Veterinary Medicine

基　　金：江南大学横向项目“动物疫苗发酵工艺优化及产业应用”。

摘　　要：【目的】以猪链球菌2型NT15B菌株为研究对象,通过优化发酵培养基组分提高菌株活菌数,降低生产成本,为猪链球菌2型NT15B菌株的高密度培养及猪链球菌病疫苗的大规模生产和推广提供参考。【方法】试验以培养的猪链球菌2型NT15B菌株最高活菌数为筛选指标,通过单因素试验筛选菌株发酵培养基中碳源、氮源(有机氮源和无机氮源)、无机盐金属离子、磷酸缓冲液的最佳组分及浓度,并根据单因素筛选结果选择碳源浓度、有机氮源总浓度、有机氮源比例、磷酸盐缓冲液浓度4个因素各3个水平设计正交试验,并进一步进行多因素的筛选与验证。【结果】猪链球菌2型NT15B菌株发酵培养基最优条件为6.0 g/L蔗糖、1.0 g/L尿素、10 g/L酵母提取物YE7与20 g/L酵母蛋白胨Nucel 887混合氮源,1.0 g/L NaCl、9.58 g/L K_(2)HPO_(4)·3H_(2)O、1.09 g/L KH_(2)PO_(4)、1 mmol/L MgSO_(4)·7H_(2)O、0.3 g/L L-抗坏血酸。采用此最优组合,摇瓶培养猪链球菌2型NT15B活菌数最高达80.83×10^(8)CFU/mL,较优化前提高256.08%。【结论】优化后的培养基能有效提高猪链球菌2型NT15B菌株活菌数,降低了生产成本,为发酵罐水平的优化提供参考。【Objective】The experiment took Streptococcus suis type 2 NT15B strain as the research object.By optimizing the fermentation medium components,the viable counts of the strain was increased and the production cost was reduced,so as to provide a reference for high-density culture of Streptococcus suis type 2 NT15B strain and the research related to the large-scale production and promotion of the vaccine against porcine streptococcal disease.【Method】The maximum viable counts of Streptococcus suis type 2 NT15B strain was used as the screening index,and the optimal components and concentrations of carbon sources,nitrogen sources(organic and inorganic nitrogen sources),inorganic salt metal ions,and phosphoric acid buffers in the fermentation medium of the strain were screened by single-factor test.According to the results of single factor screening,3 levels of each of 4 factors,carbon source concentration,total organic nitrogen source concentration,organic nitrogen source proportion and phosphate buffer concentration,were selected to design orthogonal tests,and further multi-factor screening and verification were carried out.【Result】The optimal fermentation medium of Streptococcus suis type 2 NT15B strain was 6.0 g/L sucrose,1.0 g/L urea,10 g/L yeast extract YE7 mixed with 20 g/L yeast peptone Nucel 887 as nitrogen sources,1.0 g/L NaCl,9.58 g/L K_(2)HPO_(4)·3H_(2)O,1.09 g/L KH_(2)PO_(4),1 mmol/L MgSO_(4)·7H_(2)O and 0.3 g/L L-ascorbic acid.Using this optimal combination,the viable counts of Streptococcus suis type 2 NT15B strain at the shaking flask level was 80.83×10^(8) CFU/mL,which was 256.08%higher than before optimization.【Conclusion】The optimized medium effectively increased the viable counts of Streptococcus suis type 2 NT15B strain,reduced production costs and provided a reference for optimization in fermenters.

关 键 词：猪链球菌2型 活菌数 培养基组分 优化 

分 类 号：S852.611[农业科学—基础兽医学]