响应面法优化青春双歧杆菌增殖培养基
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摘要
为提高青春双歧杆菌在发酵液中的活菌数,以MRS为培养基,采用响应面法对培养基进行优化,同时比较了优化前后的生长曲线与pH的变化。通过响应面分析结果得到青春双歧杆菌的增殖培养基配方:葡萄糖15.85g/L、低聚果糖15.85 g/L、胰蛋白胨12.04 g/L、牛肉膏7.23 g/L、酵母粉9.63 g/L、柠檬酸铵2.75 g/L、K_2HPO_4·3H_2O 2.75 g/L、乙酸钠6.875 g/L、吐温-80 1.0 mL、MgSO_4·7H_2O 0.2 g/L、MnSO_4·H_2O 0.05 g/L、L-半胱氨酸盐酸盐0.5 g/L。该菌在增殖培养基中28 h达到稳定期,比优化前缩短12 h;活菌数达到8.9×10~9CFU/mL,是优化前的1.73倍。
Response surface methodology was used to optimize the fermentation medium based on the medium MRS to increase the viable bacteria count of B.adolescentis.Meanwhile,the growth curve and pH were compared before and after optimization.According to the results of response surface analysis,the enrichment medium components were determined as follows:glucose 15.85g/L,fructo-oligosaccharide 15.85 g/L,peptone 14.45 g/L,beef extract 8.67 g/L,yeast extract power5.78 g/L,ammonium citrate 2.75 g/L,K_2HPO_4·3H_2O 2.75 g/L,sodium acetate 6.875 g/L,Tween-801.0 mL,Mg SO_4·7H_2O 0.2 g/L,MnSO_4·H_2O 0.05 g/L,L-cystein-HCl 0.5 g/L.Cultivated in optimized enrichment medium,B.adolescentis reached stationary phase at about 28 h,which was shorter 12 h than before;the viable counts reached up to 8.9×10~9CFU/mL,which was 1.73 times as many as before.
Response surface methodology was used to optimize the fermentation medium based on the medium MRS to increase the viable bacteria count of B.adolescentis.Meanwhile,the growth curve and pH were compared before and after optimization.According to the results of response surface analysis,the enrichment medium components were determined as follows:glucose 15.85g/L,fructo-oligosaccharide 15.85 g/L,peptone 14.45 g/L,beef extract 8.67 g/L,yeast extract power5.78 g/L,ammonium citrate 2.75 g/L,K_2HPO_4·3H_2O 2.75 g/L,sodium acetate 6.875 g/L,Tween-801.0 mL,Mg SO_4·7H_2O 0.2 g/L,MnSO_4·H_2O 0.05 g/L,L-cystein-HCl 0.5 g/L.Cultivated in optimized enrichment medium,B.adolescentis reached stationary phase at about 28 h,which was shorter 12 h than before;the viable counts reached up to 8.9×10~9CFU/mL,which was 1.73 times as many as before.
引文
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[3]ZHANG Ling.Development of studies on Bifldobacterium’s effects[J].Medical Recapitulate,2009,15(17):2556,2721.(in Chinese)
[4]ZHU Junchen,LI Shimin,WEI Xiaoqing.Physiological function and research progress of the human body by the Bifidobacterium[J].Chinese Journal of Microecology,2000,12(1):53-55.(in Chinese)
[5]SUN Lili,WANG Chaoqun.Isolation and identification of Bifidobacterium[J].Academic Periodical of Farm Products Processing,2011(7):99-101.(in Chinese)
[6]PERES C M,PERES C,HERNANDEZ-MENDOZA A,et al.Review on fermented plant materials as carriers and sources of potentially probiotic lactic acid bacteria-With an emphasis on table olives[J].Trends in Food Science&Technology,2012,26(1):31-42.
[7]DU Peng,LI Aili,HUO Guichheng.Optimization of medium for Bifidobacterium infantis[J].Industrial Microbiology,2005,35(3):14-18.(in Chinese)
[8]DU Shaoping,LIANG Shuwa,YANG Guandong,et al.Optimization of Bifidobacterium bifidum ferrnentation medium using response surface analysis[J].Biotechnology Bulletin,2008,24(C1):431-434.(in Chinese)
[9]SUN Peng,ZHAO Xubo,SUN Xianfeng,et al.Optimization of enrichment medium for bifidobacterium longum by response surface methodology[J].Food Science,2013,34(5):207-212.(in Chinese)
[10]XU Haiyan,ZHANG Zhiyan,CAO Bin,et al.Optimization of Bifidobacterium adolescentis basic medium and screening of freeze-drying protective agents[J].Animal Husbandry and Feed Science,2013,34(3):15-18.(in Chinese)
[11]ANVARI M,KHAYATI G,ROSTAMI S.Optimization of medium composition for probiotic biomass production using response surface methodology[J].Journal of Dairy Research,2014,81(1):59-64.
[12]CHONG Ke,TIAN Hongtao,SANG YAxin,et al.Optimization of reproducible wort medium for bifidobacteria[J].Food Science,2007,28(3):178-182.(in Chinese)
[13]HAN Xue,ZHANG Lanwei.The Bifldobacterium’s growth-promoting factors selection and the basic medium optimization[J].Journal of Food Science and Biotechnology,2005,24(4):69-72.(in Chinese)
[14]GUAN Haibin,SHI Ruiwen,DONG Zhiheng,et al.Optimizing fermentation medium of Bifidobacterium longum[J].Chinese Journal of Microecology,2014,26(3):253-257.(in Chinese)
[15]MENG Xiangchen,WANG Yafeng,HUO Guicheng.The optimization of reproducible medium for Bifidobacterium adolescent[J].Food and Fermentation Industries,2002,28(7):15-19.(in Chinese)
[16]OUYANG Wenshan,SANG Yue,LIU Li,et al.Optimization of culture medium of Bifidobacterium animalis subsp.Lactis A6Strain[J].China Dairy Cattle,2015(21):32-35.(in Chinese)
[17]SHAO Jianning,ZHANG Wenqi,MA Heping,et al.Medium optimization and pH control fermentation of Bifidobacterium[J].China Brewing,2010,29(3):122-124.(in Chinese)
[18]MEENA G S,MAJUMDAR G C,BANERJEE R,et al.Growth characteristics modeling of mixed culture of Bifidobacterium bifidum and Lactobacillus acidophilus using response surface methodology and artificial neural network[J].Brazilian Archives of Biology and Technology,2014,57(6):962-970.
[19]ABDUL K K,MUSTAFA S,MOHAMMAD R,et al.Optimization of milk-based medium for efficient cultivation of Bifidobacterium pseudocatenulatum G4 using face-centered central composite-response surface methodology[J].Biomed Reseaech International,2014,2014:78-89.
[20]中华人民共和国卫生部.GB/T 4789.34-2010食品安全国家标准食品微生物学检验乳酸菌检验[S].北京:中国标准出版社,2010.
[21]FANG Xiao.Effects of oligosaccharides on the bioactivity of Lactobacillus[J].Pharmaceutical Biotechnology,2014,21(4):338-342.(in Chinese)
[22]LAN Hekui,REN Daming.Research Progress on biological function of Bifidobacterium[J].Journal of Microbes and Infections,2002,25(3):18-20.(in Chinese)