植物乳杆菌K25发酵产胞外多糖的影响因素及其应用
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摘要
植物乳杆菌胞外多糖(extracellular polysaccharide,EPS)可赋予发酵乳制品独特的质构和风味,还具有增强免疫力、抗肿瘤和调节肠道菌群平衡等生物活性,在食品和医药领域具有良好的应用前景。但是,植物乳杆菌EPS的产量偏低,而影响其产量的因素具有菌株特异性,有关添加植物乳杆菌EPS对产品稳定性影响的研究也较少。采用单因素实验和响应面法确定了植物乳杆菌K25产EPS的优化培养基组成:每升含胰蛋白胨10 g、酵母浸粉5 g、葡萄糖20 g、磷酸氢二钾2.0 g、无水乙酸钠5.0 g、柠檬酸钠5.0 g、硫酸镁0.2 g、硫酸锰0.05 g和吐温80 1.0 mL。优化的发酵条件:发酵温度30℃、发酵时长24 h和培养基初p H值6.5。在此条件下EPS产量为250.21 mg/L。流变学研究表明,该EPS水溶液的黏度具有随着浓度和剪切速率升高而升高,以及剪切变稠的特性。扫描电镜观察发现,该EPS具有长方形块状微观结构,有利于稳定溶液的质构并保持水分。将该EPS加入到酸性乳饮料中,进一步验证了其具有良好的保水特性,表明植物乳杆菌K25产生的EPS可以作为一种具有潜在应用价值的新型食品稳定剂。
The extracellular polysaccharide( EPS) from Lactobacillus plantarum plays important roles in the special texture and flavor of fermented dairy products. These EPSs possess biological activities such as immunomodulation,antitumor and regulating the balance of intestinal flora and so on,which can be used in food and medicine fields. However,the yield of Lactobacillus plantarum EPS is relatively low,factors affecting the EPS production are strain specific,and the influence of Lactobacillus plantarum EPS on the stability of the products is still rarely studied. The optimal fermentation conditions obtained by single factor experiments and response surface analysis were as follows: peptone 10 g/L,yeast extract powder5 g/L,glucose 20 g/L,potassium hydrogen phosphate 2. 0 g/L,anhydrous sodium acetate 5. 0 g/L,sodium citrate 5. 0 g/L,magnesium sulfate 0. 2 g/L,manganese sulfate 0. 05 g/L,twain 80 1. 0 mL/L,fermentation temperature 30 ℃,fermentation time 24 h,and initial p H of culture medium of 6. 5. The maximal production of EPS was 250. 21 mg/L under the above conditions. The results of rheological characteristics studies indicated that the EPS aqueous solution exhibited the increased viscosity as the concentration of the EPS and the share rate increased,and shear thickening properties. The scanning electron microscopy verified the cuboid microstructure of the EPS molecules,facilitating stabilization of the texture and water retention in solution. Application of this EPS for preparation of acid milk beverage showed that the use of the EPS could improve stability of the beverage,indicating that the EPS from Lactobacillus plantarum K25 can be potentially used as a novel food stabilizer.
The extracellular polysaccharide( EPS) from Lactobacillus plantarum plays important roles in the special texture and flavor of fermented dairy products. These EPSs possess biological activities such as immunomodulation,antitumor and regulating the balance of intestinal flora and so on,which can be used in food and medicine fields. However,the yield of Lactobacillus plantarum EPS is relatively low,factors affecting the EPS production are strain specific,and the influence of Lactobacillus plantarum EPS on the stability of the products is still rarely studied. The optimal fermentation conditions obtained by single factor experiments and response surface analysis were as follows: peptone 10 g/L,yeast extract powder5 g/L,glucose 20 g/L,potassium hydrogen phosphate 2. 0 g/L,anhydrous sodium acetate 5. 0 g/L,sodium citrate 5. 0 g/L,magnesium sulfate 0. 2 g/L,manganese sulfate 0. 05 g/L,twain 80 1. 0 mL/L,fermentation temperature 30 ℃,fermentation time 24 h,and initial p H of culture medium of 6. 5. The maximal production of EPS was 250. 21 mg/L under the above conditions. The results of rheological characteristics studies indicated that the EPS aqueous solution exhibited the increased viscosity as the concentration of the EPS and the share rate increased,and shear thickening properties. The scanning electron microscopy verified the cuboid microstructure of the EPS molecules,facilitating stabilization of the texture and water retention in solution. Application of this EPS for preparation of acid milk beverage showed that the use of the EPS could improve stability of the beverage,indicating that the EPS from Lactobacillus plantarum K25 can be potentially used as a novel food stabilizer.
引文
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[21]YILMAZ M,CELIK G Y,ASLIM B,et al.Influence of carbon sources on the production and characterization of the exopolysaccharide(EPS)by Bacillus sphaericus7055 strain[J].Journal of Polymers and the Environment,2012,20(1):152-156.
[22]NOCELLI N,BOGINO P C,BANCHIO E,et al.Roles of extracellular polysaccharides and biofilm formation in heavy metal resistance of rhizobia[J].Materials,2016,9(6):418.
[23]HAN Y Z,LIU E Q,LIU L S,et al.Rheological,emulsifying and thermostability properties of two exopolysaccharides produced by Bacillus amyloliquefaciens LPL061[J].Carbohydrate Polymers,2015,115:230-237.
[24]AHMED Z,WANG Y P,ANJUM N,et al.Characterization of new exopolysaccharides produced by coculturing of L.kefiranofaciens with yoghurt strains[J].International Journal of Biological Macromolecules,2013,59:377-383.
[25]LAWS A P,MARSHAL V M.The relevance of exopolysaccharides to the rheological properties in milk fermented with ropy strains of lactic acid bacteria[J].International Dairy Journal,2001,11(9):709-721.
[2]TANG W,XING Z,LI C,et al.Molecular mechanisms and in vitro antioxidant effects of Lactobacillus plantarum MA2[J].Food Chemistry,2017,221:1642-1649.
[3]ZHANG Z,LIU Z,TAO X,et al.Characterization and sulfated modification of an exopolysaccharide from Lactobacillus plantarum ZDY2013 and its biological activities[J].Carbohydrate Polymers,2016,153:25-33.
[4]WANG K,LI W,RUI X,et al.Chemical modification,characterization and bioactivity of a released exopolysaccharide(r-EPS1)from Lactobacillus plantarum 70810[J].Glycoconjugate Journal,2015,32:17-27.
[5]WANG J,ZHAO X,YANG Y W,et al.Characterization and bioactivities of an exopolysaccharide produced by Lactobacillus plantarum YW32[J].International Journal of Biological Macromolecules,2015,74:119-126.
[6]WANG J,ZHAO X,TIAN Z,et al.Characterization of an exopolysaccharide produced by Lactobacillus plantarum YW11 isolated from Tibet Kefir[J].Carbohydrate Polymers,2015,125:16-25.
[7]BOELS I C,VAN KRANENBURG R,HUGENHOLTZ J,et al.Sugar catabolism and its impact on the biosynthesis and engineering of exopolysaccharide production in lactic acid bacteria[J].International Dairy Journal,2001,11(9):723-732.
[8]胡盼盼,宋微,单毓娟,等.影响乳酸菌胞外多糖产量的因素[J].食品科技,2014,39(9):31-37.HU P P,SONG W,SHAN Y J,et al.Factors contributing to the yield of exopolysaccharide by lactic acid bacteria[J].Food Science and Technology,2014,39(9):31-37.
[9]李盛钰,曾宪鹏,杨贞耐.提高乳酸菌胞外多糖产量的途径[J].食品与生物技术学报,2009,28(3):289-293.LI S Y,ZENG X P,YANG Z N.Strategies for increasing of exopolysaccharide productionin lactic acid bacteria[J].Journal of Food Science and Biotechnology,2009,28(3):289-293.
[10]MAJUMDER A,SINGH A,GOYAL A.Application of response surface methodology for glucan production from Leuconostoc dextranicum and its structural characterization[J].Carbohydrate Polymers,2009,75(1):150-156.
[11]ROMLING U,GALPERIN M Y,GOMELSKY M.Cyclic di-GMP:the first 25 years of a universal bacterial second messenger[J].Microbiology and Molecular Biology Reviews,2013,77(1):1-52.
[12]ZHANG L,ZHANG X,LIU C,et al.Manufacture of Cheddar cheese using probiotic Lactobacillus plantarum K25 and its cholesterol-lowering effects in a mice model[J].World Journal of Microbiology&Biotechnology,2013,29(1):127-135.
[13]王辑,王影,张雪,等.植物乳杆菌K25发酵乳对衰老模型小鼠的抗氧化作用[J].吉林农业大学学报,2013,35(1):98-101.WANG J,WANG Y,ZHANG X,et al.Antioxidative effect of Lactobacillus plantarum K25 fermented milk on aged model mice[J].Journal of Jilin Agricultural University,2013,35(1):98-101.
[14]王辑,张雪,李达,等.植物乳杆菌K25的技术特性[J].食品与生物技术学报,2012,31:518-524.WANG J,ZHANG X,LI D,et al.Technological properties of Lactobacillus plantarum K25[J].Journal of Food Science and Biotechnology,2012,31:518-524.
[15]郭夏蕾,张健,杨贞耐.抑制口腔变异链球菌的乳酸菌筛选及其抑菌机理[J].食品科学,2016,37(19):117-122.GUO X L,ZHANG J,YANG Z N.Screening of lactic acid bacteria for inhibiting oral Streptococcus mutans and preliminary study of antibacterial mechanism[J].Food Science,2016,37(19):117-122.
[16]DUBOC P,MOLLET B.Applications of exopolysaccharides in the dairy industry[J].International Dairy Journal,2001,11(9):759-768.
[17]YEESANG C,CHANTHACHUM S,CHEIRSILP B.Sago starch as a low-cost carbon source for exopolysaccharide production by Lactobacillus kefiranofaciens[J].World Journal of Microbiology&Biotechnology,2008,24(7):1195-1201.
[18]POLAK-BERECKA M,CHOMA A,WASKO A,et al.Physicochemical characterization of exopolysaccharides produced by Lactobacillus rhamnosus on various carbon sources[J].Carbohydrate Polymers,2015,117:501-509.
[19]TORINO M I,TARANTO M P,SESMA F,et al.Heterofermentative pattern and exopolysaccharide production by Lactobacillus helveticus ATCC 15807 in response to environmental p H[J].Journal of Applied Microbiology,2001,91(5):846-852.
[20]QIN Q L,LI Y,SUN M L,et al.Comparative transcriptome analysis reveals that lactose acts as an inducer and provides proper carbon sources for enhancing exopolysaccharide yield in the deep-sea bacterium Zunongwangia profunda SM-A87[J].Plos One,2015,10(2):e0115998.
[21]YILMAZ M,CELIK G Y,ASLIM B,et al.Influence of carbon sources on the production and characterization of the exopolysaccharide(EPS)by Bacillus sphaericus7055 strain[J].Journal of Polymers and the Environment,2012,20(1):152-156.
[22]NOCELLI N,BOGINO P C,BANCHIO E,et al.Roles of extracellular polysaccharides and biofilm formation in heavy metal resistance of rhizobia[J].Materials,2016,9(6):418.
[23]HAN Y Z,LIU E Q,LIU L S,et al.Rheological,emulsifying and thermostability properties of two exopolysaccharides produced by Bacillus amyloliquefaciens LPL061[J].Carbohydrate Polymers,2015,115:230-237.
[24]AHMED Z,WANG Y P,ANJUM N,et al.Characterization of new exopolysaccharides produced by coculturing of L.kefiranofaciens with yoghurt strains[J].International Journal of Biological Macromolecules,2013,59:377-383.
[25]LAWS A P,MARSHAL V M.The relevance of exopolysaccharides to the rheological properties in milk fermented with ropy strains of lactic acid bacteria[J].International Dairy Journal,2001,11(9):709-721.