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
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摘要
研究解淀粉芽孢杆菌JN4的胞外多糖(exopolysaccharide,EPS-JN4)对乳酸菌生长及代谢的调控作用,有利于阐明EPS-JN4定向增殖乳酸菌的作用机制。采用PCR-变性梯度凝胶电泳技术分析EPS-JN4的定向增殖作用,并考察其对主要乳酸菌生长、主要代谢产物、肠道耐受性和表面疏水性的影响。EPS-JN4可以定向增殖罗伊氏乳杆菌,其对罗伊氏乳杆菌JN125的前期增殖速度较葡萄糖慢,但最高菌浓为葡萄糖的5. 25倍,代谢产物中乳酸和醋酸含量则较葡萄糖分别降低了12. 2%和32. 4%。EPS-JN4增殖的罗伊氏乳杆菌JN125的肠道耐受性和表面疏水性显著提高,对其原因初步分析表明细胞膜蛋白和脂类物质含量提高了40. 8%和105. 7%,细胞膜组成中的十八碳脂肪酸和不饱和脂肪酸显著提高。EPS-JN4可以作为潜在的益生元,用于调节动物肠道健康。
This study aimed to investigate the effects of exopolysaccharides from Bacillus amyloliquefaciens JN4( EPS-JN4) on the propagation and metabolism of probiotics. The specific proliferative effects of EPS-JN4 were analyzed by PCR-denaturing gradient gel electrophoresis,and their effects on the growth,metabolites,intestinal tolerance,and surface hydrophobicity of lactic acid bacteria were also determined. It was found that EPS-JN4 had a specific proliferative effect on Lactobacillus reuteri. Compared to glucose,the early proliferation rate of L. reuteri JN125 treated with EPS-JN4 was slower,but its highest biomass was 4. 25 times higher. In addition,the contents of lactic acid and acetic acid reduced by 12. 2% and 32. 4%,respectively. The intestinal tolerance and surface hydrophobicity of EPS-JN4-treated L. reuteri JN125 significantly improved,which could be due to the protein and lipid contents in cell membranes increased by 40. 8% and 105. 7%,respectively. Besides,the proportions of 18 C fatty acids and unsaturated fatty acids in cell membranes significantly increased. In conclusion,EPS-JN4 can be used as a potential prebiotic to regulate the intestinal health of animals.
This study aimed to investigate the effects of exopolysaccharides from Bacillus amyloliquefaciens JN4( EPS-JN4) on the propagation and metabolism of probiotics. The specific proliferative effects of EPS-JN4 were analyzed by PCR-denaturing gradient gel electrophoresis,and their effects on the growth,metabolites,intestinal tolerance,and surface hydrophobicity of lactic acid bacteria were also determined. It was found that EPS-JN4 had a specific proliferative effect on Lactobacillus reuteri. Compared to glucose,the early proliferation rate of L. reuteri JN125 treated with EPS-JN4 was slower,but its highest biomass was 4. 25 times higher. In addition,the contents of lactic acid and acetic acid reduced by 12. 2% and 32. 4%,respectively. The intestinal tolerance and surface hydrophobicity of EPS-JN4-treated L. reuteri JN125 significantly improved,which could be due to the protein and lipid contents in cell membranes increased by 40. 8% and 105. 7%,respectively. Besides,the proportions of 18 C fatty acids and unsaturated fatty acids in cell membranes significantly increased. In conclusion,EPS-JN4 can be used as a potential prebiotic to regulate the intestinal health of animals.
引文
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[21]邱然,陆健.啤酒污染菌的鉴定及其细胞膜脂肪酸的组成分析[J].食品与发酵工业,2017,43(7):55-61.
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[23] ZHANG B,WANG Y P,TAN Z F,et al. Screening of probiotic activities of Lactobacilli strains isolated from traditional Tibetan Qula,a raw yak milk cheese[J]. AsianAustralasian Journal of Animal Sciences,2016,29(10):1 490-1 499.
[24] SUZUKI K,IIJIMA K,SAKAMOTO K,et al. A review of hop resistance in beer spoilage lactic acid bacteria[J].Journal of the Institute of Brewing,2006,112(2):173-191.
[25] MYKYTCZUK N C S,TREYORS J T,LEDUC L G,et al. Fluorescence polarization in studies of bacterial cytoplasmic membrane fluidity under environmental stress[J]. Progress in Biophysics and Molecular Biology,2007,95(3):60-82.
[2] ONER E T,HEMANDEZ L,COMBIE J. Review of Levan polysaccharide:From a century of past experiences to future prospects[J]. Biotechnology Advances,2016,34(5):827-844.
[3] SCHMID J. Recent insights in microbial exopolysaccharide biosynthesis and engineering strategies[J]. Current Opinion in Biotechnology,2018,53:130-136.
[4] LIANG T W,WANG S L. Recent advances in exopolysaccharides from Paenibacillus spp.:Production,isolation,structure,and bioactivities[J]. Marine Drugs,2015,13(4):1 847-1 863.
[5] BADEL S,BERNARDI T,MICHAUD P. New perspectives for Lactobacilli exopolysaccharides[J]. Biotechnology Advance,2011,29(1):54-66.
[6] FELS L,JAKOB F,VOGEL R F,et al. Structural characterization of the exopolysaccharides from water kefir[J].Carbohydrate Polymers,2018,189:296-303.
[7] ZHANG H,REN W,GUO Q B,et al. Characterization of a yogurt-quality improving exopolysaccharide from Streptococcus thermophilus AR333[J]. Food Hydrocolloids,2018,81,220-228.
[8] YU X,AVALL-JAASKELAINEN S,KOOT J,et al. A comparative characterization of different host-sourced Lactobacillus ruminis strains and their adhesive,inhibitory,and immunomodulation functions[J]. Frontiers in Microbiology,2017,8:657.
[9] FUCHS V I,SCHMIDT J,SLATER M J,et al. Influence of immunostimulant polysaccharides,nucleic acids,and Bacillus strains on the innate immune and acute stress response in turbots(Scophthalmus maximus)fed soy beanand wheat-based diets[J]. Fish Physiology and Biochemistry,2017,43(6):1 501-1 515.
[10] POURABEDIN M,ZHAO X. Prebiotics and gut microbiota in chickens[J]. FEMS Microbiology Letters,2015,362(15):fnv122.
[11] GOULET O. Potential role of the intestinal microbiota in programming health and disease[J]. Nutrition Reviews,2015,73(Supp 1):32-40.
[12] SALAZAR N, GUEIMONDE M, DW LOS REYESGAVILN C G,et al. Exopolysaccharides produced by Lactic acid bacteria and Bifidobacteria as fermentable substrates by the intestinal microbiota[J]. Critical Reviews in Food Science and Nutrition,2016,56(9):1 440-1 453.
[13] GOMAA E Z. Effect of prebiotic substances on growth,fatty acid profile and probiotic characteristics of Lactobacillus brevis NM101-1[J]. Microbiology,2017,86(5):618-628.
[14] MOZZI F,GERBINO E,DE VADEZ G F,et al. Functionality of exopolysaccharides produced by lactic acid bacteria in an in vitro gastric system[J]. Journal of Applied Microbiology,2009,107(1):56-64.
[15] TOLSTOGUZOV V. Why were polysaccharides necessary?[J]. Origins of Life and Evolution of Biospheres,2004,34(6):571-597.
[16]蔡国林,冯文旭,刘逸凡,等.高产抑制大肠杆菌血凝性的胞外多糖的解淀粉芽孢杆菌[J].食品与发酵工业,2019,45(5):14-18.
[17] JACOBS C M,UTTERBACK P L,PARSONS C M. Cecal microbial populations of young chicks fed several prebiotic-type compounds as determined by DGGE and quantitative PCR[J]. Journal of Dairy Science,2010,93:283-283.
[18] GOMAA E Z. Effect of prebiotic substances on growth,fatty acid profile and probiotic characteristics of Lactobacillus brevis NM101-1[J]. Microbiology,2017,86(5):618-628.
[19] DAS D,BARUAH R,GOYAL A. A food additive with prebiotic properties of anα-d-glucan from Lactobacillus plantarum DM5[J]. International Journal of Biological Macromolecules,2014,69:20-26.
[20]杨亚威,赵爱梅,王辑,等.传统奶豆腐中产胞外多糖乳酸菌的分离筛选及其潜在益生菌特性[J].中国乳品工业,2015,43(12):8-13.
[21]邱然,陆健.啤酒污染菌的鉴定及其细胞膜脂肪酸的组成分析[J].食品与发酵工业,2017,43(7):55-61.
[22] DE PRETER V,HAMER H M,WINDEY K,et al. The impact of pre-and/or probiotics on human colonic metabolism:Does it affect human health?[J]. Molecular Nutrition and Food Research,2011,55(1):46-57.
[23] ZHANG B,WANG Y P,TAN Z F,et al. Screening of probiotic activities of Lactobacilli strains isolated from traditional Tibetan Qula,a raw yak milk cheese[J]. AsianAustralasian Journal of Animal Sciences,2016,29(10):1 490-1 499.
[24] SUZUKI K,IIJIMA K,SAKAMOTO K,et al. A review of hop resistance in beer spoilage lactic acid bacteria[J].Journal of the Institute of Brewing,2006,112(2):173-191.
[25] MYKYTCZUK N C S,TREYORS J T,LEDUC L G,et al. Fluorescence polarization in studies of bacterial cytoplasmic membrane fluidity under environmental stress[J]. Progress in Biophysics and Molecular Biology,2007,95(3):60-82.
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