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China Feed
2019
(7)
19-27
粪肠球菌R-WL发酵培养基的优化
作 者：
陈国参;王佰涛;徐文涛;王一雯;李寒冰;刘德海
单 位：
河南省科学院生物研究所有限责任公司
关键词：
粪肠球菌R-WL;发酵培养基;Plackett-Burman;Box-Behnken;响应面分析
摘 要：
本试验旨在优化培养基提高粪肠球菌(Enterococcus faecalis)R-WL发酵液的菌体密度,为高效、优质的微生态制剂产品研制奠定基础。首先通过生长曲线的测定、培养条件和培养基成分的单因素试验,确定菌种RWL的最适培养条件为:时间14 h、pH 8.5、温度37℃、接种量3%,最适碳源为乳糖,浓度为10 g/L,最适氮源为蛋白胨+酵母膏,浓度为30 g/L。然后通过Plackett-Burman试验分析培养基中对粪肠球菌发酵影响最重要的主要因素为蛋白胨、乙酸钠和柠檬酸二铵,再通过最陡爬坡试验获得主要因素的最适范围,最后通过Box-Behnken试验和响应面分析得到主要因素的最适添加量。结果表明:最优发酵培养基配方为:乳糖10 g/L、酵母膏15 g/L、蛋白胨12.18 g/L、乙酸钠4.5 g/L、柠檬酸二铵4.5 g/L、硫酸镁0.58 g/L、硫酸锰0.25 g/L、磷酸氢二钾2 g/L、轻质碳酸钙3 g/L,初始pH 8.5。通过试验验证,优化后发酵培养基的发酵菌液活菌数可达(6.44±0.10)×109cfu/mL,比原MRS培养基活菌数提高了71.28%。
译 名：
Optimization of fermentation medium for Enterococcus faecalis R-WL
作 者：
HU Hongwei;MA Xiaotao;YAN Lingpeng;CHEN Ruru;ZHANG Chanjuan;YANG Jinge;LI Ziru;DANG Yapeng;REN Shuai;Shanxi Dayu Biological Engineering Co.,Ltd;
单 位：
CHEN Guocan%WANG Baitao%XU Wentao%WANG Yiwen%LI Hanbing%LIU Dehai%Institute of Biology,Henan Academy of Science%Henan Industrial Enzyme Engineering Technology Research Eenter
关键词：
Enterococcus faecalis R-WL;;fermentation medium;;Plackett-Burman;;Box-Behnken;;response surface methodology
摘 要：
This experiment aimed to improve the concentration of Enterococcus faecalis R-WL by optimizing fermentation medium,and to lay the foundation for efficient quality probiotics.First,measured the growth curve assay and single factor test for culture condition and medium composition.The optimum incubation time,pH,temperature,inoculation amount of R-WL were 14 h,8.5,37 ℃,3%,the optimum carbon and nitrogen sources were lactose and peptone + yeast extract,the concentration were 10 g/L and 30 g/L,respectively.Plackett-Burman test was applied to analyze the main factors of fermentation medium were peptone,sodium acetate and diammonium citrate,and then the main factors were tested through the steepest ascent method to get the optimal range.Finally,the optimum addition of main factors was obtained by Box-Behnken test and the response surface methodology. The optimal formulation of fermentation medium were:lactose 10 g/L,yeast extract15 g/L,peptone 12.18 g/L,sodium acetate 4.5 g/L,diammonium citrate 4.5 g/L,magnesium sulphate 0.58 g/L,manganese sulfate 0.25 g/L,potassium hydrogen phosphate 2 g/L,light calcium carbonate 3 g/L,pH 8.5.Enterococcus faecalis were cultured by the optimized fermentation medium,and the viable count reached(6.44±0.10)×109 cfu/mL,which increased by 71.28% compared with the number of viable count in MRS culture.
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