为提高双歧杆菌BB12胞外多糖(exopolysaccharide,EPS)的产量,并探究其抗氧化活性,该文通过单因素试验分析初始pH值、培养温度和培养时间对胞外多糖产量的影响,在此基础上通过响应面法进一步优化发酵工艺参数,并对胞外多糖的还原能力、·OH和·O-2的清除作用进行了研究。结果表明,最优发酵条件修正后为:初始pH值8、培养温度37 ℃、培养时间101 h。在此条件下进行发酵验证,得到的胞外多糖产量为(131.6±0.82) mg/L。体外抗氧化性试验表明双歧杆菌胞外多糖具有还原能力,对·OH和·O-2的50%清除浓度(IC50)均低于抗坏血酸(Vc)。为双歧杆菌胞外多糖的大规模生产及抗氧化机制的研究提供了可靠的理论依据。
The purpose of this study was to enhance exopolysaccharide (EPS) yield from Bifidobacterium lactis BB12® (BB12) and to determine the antioxidant activity of EPS. Single factor experiment was used to analyze the effects of initial pH, temperature and time on the yield of EPS. Subsequently, the fermentation parameters were further improved using response surface methodology. The antioxidant activity was determined by ferric reducing power of EPS, and the -OH and -O2 scavenging rate. The results showed that initial pH of 8, temperature of 37 ℃, and incubation time of 101 h was the optimum processing condition for producing EPS. Finally, the yield of EPS reached 131.6±0.82 mg/L. In addition, the reducing ability of EPS was further evaluated using in vitro antioxidant test, which IC50 -OH and -O2 was lower than that of Vc. Our research provided a reliable theoretical basis for the EPS commercial scale production using BB12, as well as foundations of understanding for antioxidant mechanism.
[1] SHANG N, XU R, LI P, et al. Structure characterization of an exopolysaccharide produced by Bifidobacterium animalis RH[J]. Carbohydrate Polymers, 2013, 91(1): 128-134.
[2] 赵建云, 左芳雷,肖满,等. 双歧杆菌耐氧性研究及其应用[J]. 食品与发酵工业, 2010, 36(12): 15-19.
[3] 王猛, 熊江,张玲,等. 动物双歧杆菌乳亚种BZ11的耐氧驯化及降胆固醇性能的研究[J]. 食品与发酵工业, 2016, 42(3): 1-7.
[4] FREITAS F, ALVES V D, REIS M A. Advances in bacterial exopolysaccharides: From production to biotechnological applications[J]. Trends Biotechnol, 2011, 29(8): 388-398.
[5] 白少峰, 陈华海,王欣,等. 双歧杆菌胞外多糖研究进展[J]. 中国微生态学杂志, 2017, 29(10): 1 207-1 211;1 218.
[6] XU R, SHEN Q, DING X, et al. Chemical characterization and antioxidant activity of an exopolysaccharide fraction isolated from Bifidobacterium animalis, RH [J]. European Food Research & Technology, 2011, 232(2):231-240.
[7] 蔡国林, 刘逸凡,李晓敏,等. 解淀粉芽孢杆菌胞外多糖对乳酸菌生长及代谢的调控作用[J]. 食品与发酵工业, 2019, 45(10): 16-21.
[8] HIDALGO C C, LAPEZ P, GUEIMONDE M, et al. Immune modulation capability of exopolysaccharides synthesised by lactic acid bacteria and bifidobacteria[J]. Probiotics & Antimicrobial Proteins, 2012, 4(4):227-237.
[9] WU M H, PAN T M, WU Y J, et al. Exopolysaccharide activities from probiotic bifidobacterium: Immunomodulatory effects (on J774A.1 macrophages) and antimicrobial properties [J]. International Journal of Food Microbiology, 2010, 144(1): 104-110.
[10] LI W, TANG W, JI J, et al. Characterization of a novel polysaccharide with anti-colon cancer activity from Lactobacillus helveticus MB2-1[J]. Carbohydrate Research, 2015, 411:6-14.
[11] WANG K, LI W, RUI X, et al. Characterization of a novel exopolysaccharide with antitumor activity from Lactobacillus plantarum 70810[J]. International Journal of Biological Macromolecules, 2014, 63:133-139.
[12] RUSSELL D A, ROSS R P, GF, et al. Metabolic activities and probiotic potential of bifidobacterial[J]. Int J Food Microbiol, 2011, 149(1): 88-105.
[13] 陈钧辉. 生物化学实验[M]. 北京:科学出版社, 2015.
[14] WANG S L, LIU K C, LIANG T W, et al. In vitro antioxidant activity of liquor and semi-purified fractions from squid pen biowaste by Serratia ureilytica TKU013[J]. Food Chem, 2010, 119: 1 380-1 385.
[15] YANG H, DENG J, YUAN Y, et al. Two novel exopolysaccharides from Bacillus amyloliquefaciens C-1: antioxidation and effect on oxidative stress[J]. Current Microbiology, 2015, 70(2): 298-306.
[16] 刘逸凡, 蔡国林,李晓敏,等. 丁酸梭菌的筛选及其胞外多糖抗氧化性的研究[J]. 食品与发酵工业, 2019, 45(5): 25-30.
[17] 袁建锋, 蔡恒,单咸旸,等. 一株芽孢杆菌胞外多糖的分离纯化及其抗氧化性测定[J]. 微生物学通报, 2009, 36(10): 1 466-1 470.
[18] 许冰. 一株芽孢杆菌胞外多糖的抗氧化性研究[J]. 中国酿造, 2010(8): 75-76.
[19] LIANG T W, TSENG S C, WANG S L. Production and characterization of antioxidant properties of exopolysaccharide(s) from Peanibacillus mucilaginosus TKU032[J]. Marine Drugs, 2016, 14(40): 1-12.
