利用响应面法优化发酵乳杆菌AR497电转化条件  

Optimization of electroporation method of Lactobacillus fermentum AR497 using response surface methodology

作　　者：韦云莹 赵燕 庄金伟 李江 帕米拉.乃则木丁 陆祎晟 艾连中[1] 熊智强[1] WEI Yunying;ZHAO Yan;ZHUANG Jinwei;LI Jiang;Pamila NAIZEMUDING;LU Yisheng;AI Lianzhong;XIONG Zhiqiang(Shanghai Engineering Research Center of Food Microbiology,School of Medical Instrument andFood Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China)

机构地区：[1]上海理工大学医疗器械与食品学院上海食品微生物工程研究中心,上海200093

出　　处：《食品与发酵工业》2019年第7期52-58,共7页Food and Fermentation Industries

基　　金：国家自然科学基金(31871776;31771956);十三五国家重点研发计划(2018YFD0502306);上海市自然科学基金(18ZR1426800);上海市曙光计划项目(15SG42)

摘　　要：以1株能有效改善小鼠肠炎的发酵乳杆菌AR497为研究对象,在Plackett-Burman试验的基础上选取甘氨酸质量浓度、OD_(600)、电场强度和山梨醇浓度为自变量,电转化效率为响应值,利用Box-Behnken响应面法优化其电转化条件。最优条件为:电场强度15 kV/cm,山梨醇浓度0.6 mol/L,OD_(600)0.2,甘氨酸质量浓度26 g/L。在此条件下,电转化效率高达2.78×10~5 CFU/μg DNA,比优化前提高了22倍,实现了发酵乳杆菌AR497的高效遗传转化,为后续对其进行遗传改造和解析抗炎机制奠定基础。Lactobacillus fermentum AR497 that can evidently relieve intestinal inflammation in mice was used as the main subject. Based on the Plackett-Burman tests, glycine concentrations, OD600, electric field strength, and sorbitol concentrations were selected as independent variables, the electroporation efficiency was the response value. Further optimization was made by using Box-Behnken response surface methodology. The optimum values for electric field strength,sorbitol content,OD600, and glycine content were 15 kV/cm,0.6 mol/L,0.2 and 26 g/L, respectively. The transformants reached 2.78×10^5 CFU/μg DNA under the above conditions, which was improved about 22-fold compared with that of unoptimized condition. The highly frequent genetic transformation of AR497 was realized in this work, which laid the foundation for its genetic editing and elucidating its anti-inflammation mechanism.

关 键 词：发酵乳杆菌 感受态细胞 电转化 响应面法 

分 类 号：TS201.3[轻工技术与工程—食品科学]