分离自传统自然发酵食品中降胆固醇乳酸菌的筛选与评价
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摘要
为了筛选优良的降胆固醇乳酸菌,本分析了从传统自然发酵食品中分离的30株乳酸菌,从中筛选出WS1、LP2、LP3、LP4、LP5和LP6 6株具有较强降胆固醇能力的乳酸菌,并通过对其胆盐耐受性及在含胆盐MRS培养基中的降胆固醇效率的分析,复筛出其中的WS1、LP2、LP3和LP6 4株乳酸菌。对4株乳酸菌的生长特征、产酸特征、不同生长阶段的种子液对胆固醇的降低效率的影响、耐酸特征及人工胃酸的耐受性进行分析和评价。结果显示,4株优选的乳酸菌降胆固醇能力较好,在胆盐中具有一定的存活力。胆盐的存在对降胆固醇能力具有一定的促进作用,4株菌在常规MRS培养基中生长和产酸特征稳定。不同生长时期的种子液对乳酸菌的降胆固醇能力有一定影响,对数期优于稳定期。耐酸性分析表明, pH越高,乳酸菌耐受性越好。pH 2.5,处理2h,乳酸菌依然具有一定的存活力;pH 3.5,乳酸菌存活力和存活时间都有所提高,pH 4.5时的存活力基本和自然pH的保持一致。在人工胃酸条件下,4株复筛乳酸菌在pH 2的条件下,存活时间超过4 h,随着pH的升高,存活率和存活时间都有所提高。本试验结果表明,4株乳酸菌均具有降胆固醇的特征,为后续优良菌株的降胆固醇机理研究打下了基础。
The aim of this study was to screen out good cholesterol-lowering lactic acid bacteria.6 strains of lactic acid bacteria,WS1,LP2,LP3,LP4,LP5 and LP6,which showed stronger cholesterol-lowering ability,were selected out from 30 strains of lactic acid bacteria isolated from traditional naturally fermented food.The tolerance of the 6 strains was analyzed and their cholesterol-degrading efficiency and effectiveness were detected in MRS medium with bile salt.Following this,4 strains,WS1,LP2,LP3 and LP6 were further sieved out.It was analyzed and evaluated for the 4 strains lactic acid bacteria about their growth curves,acid-yield feature,effect of seed from different growth stage on cholesterol-lowering ability and resistance in simulated gastric acid.The results showed that cholesterol-lowering ability of the 4 strains was very nice and could survive in certainly concentration bile salt.The cholesterol-lowering ability was promoted with the presence of bile salt.The growth curve and acid yield feature of 4 strains were stable in normal MRS medium.The cholesterol-degrading ability was different between the seed from different growth phases,the seed from log phase better than that from stable phase.In this study,the result of acid tolerance showed that the resistance of these lactic acid bacteria was stronger when pH was higher.With the pH going higher,the viability increased.Their growth hardly showed difference between pH 4.5 and normal pH.4 strains could survived 4 h in artificial gastric acid with pH2 and the viable rate and time extended with the increase of pH.The results implied that the 4 strains could be used in manufacture which needed cholesterol-lowering lactic acid bacteria as starter.It also laid foundation for the further research on cholesterol-lowering mechanism of the 4 strains lactic acid bacteria.
The aim of this study was to screen out good cholesterol-lowering lactic acid bacteria.6 strains of lactic acid bacteria,WS1,LP2,LP3,LP4,LP5 and LP6,which showed stronger cholesterol-lowering ability,were selected out from 30 strains of lactic acid bacteria isolated from traditional naturally fermented food.The tolerance of the 6 strains was analyzed and their cholesterol-degrading efficiency and effectiveness were detected in MRS medium with bile salt.Following this,4 strains,WS1,LP2,LP3 and LP6 were further sieved out.It was analyzed and evaluated for the 4 strains lactic acid bacteria about their growth curves,acid-yield feature,effect of seed from different growth stage on cholesterol-lowering ability and resistance in simulated gastric acid.The results showed that cholesterol-lowering ability of the 4 strains was very nice and could survive in certainly concentration bile salt.The cholesterol-lowering ability was promoted with the presence of bile salt.The growth curve and acid yield feature of 4 strains were stable in normal MRS medium.The cholesterol-degrading ability was different between the seed from different growth phases,the seed from log phase better than that from stable phase.In this study,the result of acid tolerance showed that the resistance of these lactic acid bacteria was stronger when pH was higher.With the pH going higher,the viability increased.Their growth hardly showed difference between pH 4.5 and normal pH.4 strains could survived 4 h in artificial gastric acid with pH2 and the viable rate and time extended with the increase of pH.The results implied that the 4 strains could be used in manufacture which needed cholesterol-lowering lactic acid bacteria as starter.It also laid foundation for the further research on cholesterol-lowering mechanism of the 4 strains lactic acid bacteria.
引文
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[10]RAS R T,KOPPENOL W P,GARCZAREK U,et al.Increases in plasma plant sterols stabilize within four weeks of plant sterol intake and are independent of cholesterol metabolism[J].Nutrition Metabolism&Cardiovascular Diseases,2015,26(4):302-309.
[11]KHANI S,HOSSEINI H M,TAHERI M,et al Probiotics as an alternative strategy for prevention and treatment of human diseases:a review[J].Inflammation&Allergy-Drug Targets,2012,11(2):79-89.
[12]GUO C F,ZHANG S,YUAN Y H,et al.Comparison of lactobacilli isolated from Chinese suantsai and koumiss for their probiotic and functional properties[J].Journal of Functional Foods,2015,12(2):294-302.
[13]FRAN覶OISE L.Occurrence and role of lactic acid bacteria in seafood products[J].Food Microbiology,2010,27(6):698-709.
[14]AKANBI B O,AGARRY O O.Hypocholesterolemic and growth promoting effects of AK isolated from a nigerian fermented cereal product on rats fed high fat diet[J].Advances in Microbiology,2014,4(3):160-166.
[15]靳志敏,通力嘎,段艳,等.1株高效降胆固醇乳酸菌的功能与发酵特性[J].食品科学,2014,35(11):120-123.
[16]HUANG W C,CHEN Y M,KAN N W,et al Hypolipidemic effects and safety of Lactobacillus reuteri 263 in a hamster model of hyperlipidemia[J].Nutrients,2015,7(5):3767-3782.
[17]KUMAR M,NAGPAL R,KUMAR R,et al Cholesterol-lowering probiotics as potential biotherapeutics for metabolic diseases[J].Experimental Diabetes Research,2012,2012(3):344-350.
[18]HUANG Y,WANG X,WANG J,et al.Lactobacillus plantarum strains as potential probiotic cultures with cholesterol-lowering activity[J].Journal of Dairy Science,2013,96(5):2746-2753.
[19]张根生,丁小君,于敏,等.复合乳酸菌发酵降低蛋黄液中胆固醇的工艺优化[J].食品科学,2016,37(3):186-190.
[20]吕秀红,陈凯飞,朱祺,等.降胆固醇乳酸菌的筛选与鉴定[J].中国食品学报,2016,16(3):198-204.
[21]JONES M L,TOMARODUCHESNEAU C,MAR-TONI C J,et al.Cholesterol lowering with bile salt hydrolase-active probiotic bacteria,mechanism of action,clinical evidence,and future direction for heart health applications[J].Expert Opinion on Biological Therapy,2013,13(5):631-642.
[22]ISHIMWE N,DALIRI E B,LEE B H,et al.The perspective on cholesterol-lowering mechanisms of probiotics[J].Molecular Nutrition&Food Research,2015,59:94-105.
[23]王秀娟,张小磊,商雪娇,等.2种乳酸菌降胆固醇能力的比较研究[J].食品科技,2016,41(5):12-15.
[24]中华人民共和国国家卫生和计划生育委员会.GB5009128--2016食品中胆固醇的测定[S].北京:中国标准出版社,2016.
[25]KUMAR A,KUMAR M,GHOSH M,et al.Modeling in vitro cholesterol reduction in relation to growth of probiotic Lactobacillus casei[J].Microbiology and Immunology,2013,57(2):100-110.
[26]GUO C F,ZHANG L W,HAN X,et al.Screening for cholesterol-lowering probiotic based on deoxycholic acid removal pathway and studying its functional mechanisms invitro[J].Anaerobe,2012,18(5):516-522.
[27]LEE N K,KIM S Y,HAN K J,et al.Probiotic potential of Lactobacillus,strains with anti-allergic effects from kimchi for yogurt starters[J].LWT-Food Science and Technology,2014,58(1):130-134.
[28]WANG J,ZHANG H,CHEN X,et al.Selection of potential probiotic lactobacilli for cholesterol-lowering properties and their effect on cholesterol metabolism in rats fed a high-lipid diet[J].Journal of Dairy Science,2012,95(4):1645-1654.
[29]HUANG Y,WU F,WANG X,et al.Characterization of Lactobacillus plantarum Lp27 isolated from Tibetan kefir grains:A potential probiotic bacterium with cholesterol-lowering effects[J].Journal of Dairy Science,2013,96(5):2816-2825.
[30]ANGMO K,KUMARI A,SAVITRI,et al.Probiotic characterization of lactic acid bacteria isolated from fermented foods and beverage of Ladakh[J].LWT-Food Science and Technology,2015,66:428-435.
[31]ZHOU F,ZHAO H,ZHENG J,et al.Evaluation of probiotic properties of Lactobacillus,strains isolated from traditional Chinese cheese[J].Annals of Microbiology,2015,65(3):1-8.
[32]PAPAMANOLI E,TZANETAKIS N,LITOPOULOU-TZANETAKI E,et al.Characterization of lactic acid bacteria isolated from a Greek dry-fermented sausage in respect of their technological and probi otic properties[J].Meat Science,2003,65(2):859-867.
[33]PITINO I,RANDAZZO C L,MANDALARI G,et al.Survival of Lactobacillus rhamnosus,strains in the upper gastrointestinal tract[J].Food Microbiology,2010,27(8):1121-1127.
[34]ILAVENIL S,PARK H S,VIJAYAKUMAR M,et al.Probiotic potential of Lactobacillus strains with antifungal activity isolated from animal manure[J].The Scientific World Journal,2015,2015:1-10.
[35]杨晶,邹积宏,关力,等.具有降胆固醇功能的益生菌的筛选[J].中国微生态学杂志,2012,24(10):297-300.
[36]郭春锋,张兰威.益生菌降胆固醇功能研究进展[J]微生物学报,2010,50(12):1590-1599.
[37]DARJANI P,NEZHAD M H,KADKHODAEE R,et al.Influence of prebiotic and coating materials on morphology and survival of a probiotic strain of Lactobacillus casei,exposed to simulated gastrointestinal conditions[J].LWT-Food Science and Technology,2016,73:162-167.
[38]PANKASEMSUK T,APICHARTSRANGKOON A,WORAMETRACHANON S,et al.Encapsulation of Lacto bacillus casei,01 by alginate along with himaize starch for exposure to a simulated gut model[J].Food Bioscience,2016,16:32-36.
[39]DIMITRELLOU D,KANDYLIS P,PETROVIC T,et al.Survival of spray dried microencapsulated Lactobacillus casei,ATCC 393 in simulated gastrointestinal conditions and fermented milk[J].LWT-Food Science and Technology,2016,71:169-174.
[40]LEONE R D S,ANDRADE E F D,ELLENDER-SEN L N,et al.Evaluation of dried yacon(Smallanthus sonchifolius)as an efficient probiotic carrier of Lactobacillus casei,LC-01[J].LWT-Food Science and Technology,2017,75:220-226.
[2]李琳.降胆固醇植物乳杆菌差异表达蛋白的筛选与鉴定[D].密山:黑龙江八一农垦大学,2015.
[3]国立东,王丽群,蒋琛,等.乳酸菌调控体内胆固醇代谢综述[J].中国乳品工业,2016,44(2):32-36.
[4]HAN Y,QI R,LIU G.Reduced high density lipoprotein cholesterol in severe hypertriglyceridemic ApoCIII transgenic mice via lowered hepatic ApoAIsynthesis[J].Biochemical&Biophysical Research Communications,2015,462(4):420-425.
[5]GLUECK C J,SHAH P,GOLDENBERG N,et al Eligibility for PCSK9 treatment in 734 Hypercholesterolemic patients referred to a regional cholesterol treatment center with LDL cholesterol≥70 mg/dl despite maximal tolerated cholesterol lowering therapy[J].Lipids in Health&Disease,2016,15(1):1-6.
[6]CORTESE F,GESUALDO M,CORTESE A,et al Rosuvastatin:Beyond the cholesterol-lowering effect[J].Pharmacological Research,2016,107:1-18.
[7]JO S Y,CHOI E A,LEE J J,et al.Characterization of starter kimchi fermented with Leuconostoc kimchii GJ2 and its cholesterol‐lowering effects in rats fed a high‐fat and high‐cholesterol diet[J].Journal of the Science of Food and Agriculture,2015,95(13):2750-2756.
[8]SURAMPUDI P,ENKHMAA B,ANUURAD E,et al.Lipid lowering with soluble dietary fiber[J].Current Atherosclerosis Reports,2016,18(75):1-13.
[9]TANSAZ S,DURMANN A,DETSCH R,et al Hydrogel films and microcapsules based on soy protein isolate combined with alginate[J].Journal of Applied Polymer Science,2016,4(20):1-9.
[10]RAS R T,KOPPENOL W P,GARCZAREK U,et al.Increases in plasma plant sterols stabilize within four weeks of plant sterol intake and are independent of cholesterol metabolism[J].Nutrition Metabolism&Cardiovascular Diseases,2015,26(4):302-309.
[11]KHANI S,HOSSEINI H M,TAHERI M,et al Probiotics as an alternative strategy for prevention and treatment of human diseases:a review[J].Inflammation&Allergy-Drug Targets,2012,11(2):79-89.
[12]GUO C F,ZHANG S,YUAN Y H,et al.Comparison of lactobacilli isolated from Chinese suantsai and koumiss for their probiotic and functional properties[J].Journal of Functional Foods,2015,12(2):294-302.
[13]FRAN覶OISE L.Occurrence and role of lactic acid bacteria in seafood products[J].Food Microbiology,2010,27(6):698-709.
[14]AKANBI B O,AGARRY O O.Hypocholesterolemic and growth promoting effects of AK isolated from a nigerian fermented cereal product on rats fed high fat diet[J].Advances in Microbiology,2014,4(3):160-166.
[15]靳志敏,通力嘎,段艳,等.1株高效降胆固醇乳酸菌的功能与发酵特性[J].食品科学,2014,35(11):120-123.
[16]HUANG W C,CHEN Y M,KAN N W,et al Hypolipidemic effects and safety of Lactobacillus reuteri 263 in a hamster model of hyperlipidemia[J].Nutrients,2015,7(5):3767-3782.
[17]KUMAR M,NAGPAL R,KUMAR R,et al Cholesterol-lowering probiotics as potential biotherapeutics for metabolic diseases[J].Experimental Diabetes Research,2012,2012(3):344-350.
[18]HUANG Y,WANG X,WANG J,et al.Lactobacillus plantarum strains as potential probiotic cultures with cholesterol-lowering activity[J].Journal of Dairy Science,2013,96(5):2746-2753.
[19]张根生,丁小君,于敏,等.复合乳酸菌发酵降低蛋黄液中胆固醇的工艺优化[J].食品科学,2016,37(3):186-190.
[20]吕秀红,陈凯飞,朱祺,等.降胆固醇乳酸菌的筛选与鉴定[J].中国食品学报,2016,16(3):198-204.
[21]JONES M L,TOMARODUCHESNEAU C,MAR-TONI C J,et al.Cholesterol lowering with bile salt hydrolase-active probiotic bacteria,mechanism of action,clinical evidence,and future direction for heart health applications[J].Expert Opinion on Biological Therapy,2013,13(5):631-642.
[22]ISHIMWE N,DALIRI E B,LEE B H,et al.The perspective on cholesterol-lowering mechanisms of probiotics[J].Molecular Nutrition&Food Research,2015,59:94-105.
[23]王秀娟,张小磊,商雪娇,等.2种乳酸菌降胆固醇能力的比较研究[J].食品科技,2016,41(5):12-15.
[24]中华人民共和国国家卫生和计划生育委员会.GB5009128--2016食品中胆固醇的测定[S].北京:中国标准出版社,2016.
[25]KUMAR A,KUMAR M,GHOSH M,et al.Modeling in vitro cholesterol reduction in relation to growth of probiotic Lactobacillus casei[J].Microbiology and Immunology,2013,57(2):100-110.
[26]GUO C F,ZHANG L W,HAN X,et al.Screening for cholesterol-lowering probiotic based on deoxycholic acid removal pathway and studying its functional mechanisms invitro[J].Anaerobe,2012,18(5):516-522.
[27]LEE N K,KIM S Y,HAN K J,et al.Probiotic potential of Lactobacillus,strains with anti-allergic effects from kimchi for yogurt starters[J].LWT-Food Science and Technology,2014,58(1):130-134.
[28]WANG J,ZHANG H,CHEN X,et al.Selection of potential probiotic lactobacilli for cholesterol-lowering properties and their effect on cholesterol metabolism in rats fed a high-lipid diet[J].Journal of Dairy Science,2012,95(4):1645-1654.
[29]HUANG Y,WU F,WANG X,et al.Characterization of Lactobacillus plantarum Lp27 isolated from Tibetan kefir grains:A potential probiotic bacterium with cholesterol-lowering effects[J].Journal of Dairy Science,2013,96(5):2816-2825.
[30]ANGMO K,KUMARI A,SAVITRI,et al.Probiotic characterization of lactic acid bacteria isolated from fermented foods and beverage of Ladakh[J].LWT-Food Science and Technology,2015,66:428-435.
[31]ZHOU F,ZHAO H,ZHENG J,et al.Evaluation of probiotic properties of Lactobacillus,strains isolated from traditional Chinese cheese[J].Annals of Microbiology,2015,65(3):1-8.
[32]PAPAMANOLI E,TZANETAKIS N,LITOPOULOU-TZANETAKI E,et al.Characterization of lactic acid bacteria isolated from a Greek dry-fermented sausage in respect of their technological and probi otic properties[J].Meat Science,2003,65(2):859-867.
[33]PITINO I,RANDAZZO C L,MANDALARI G,et al.Survival of Lactobacillus rhamnosus,strains in the upper gastrointestinal tract[J].Food Microbiology,2010,27(8):1121-1127.
[34]ILAVENIL S,PARK H S,VIJAYAKUMAR M,et al.Probiotic potential of Lactobacillus strains with antifungal activity isolated from animal manure[J].The Scientific World Journal,2015,2015:1-10.
[35]杨晶,邹积宏,关力,等.具有降胆固醇功能的益生菌的筛选[J].中国微生态学杂志,2012,24(10):297-300.
[36]郭春锋,张兰威.益生菌降胆固醇功能研究进展[J]微生物学报,2010,50(12):1590-1599.
[37]DARJANI P,NEZHAD M H,KADKHODAEE R,et al.Influence of prebiotic and coating materials on morphology and survival of a probiotic strain of Lactobacillus casei,exposed to simulated gastrointestinal conditions[J].LWT-Food Science and Technology,2016,73:162-167.
[38]PANKASEMSUK T,APICHARTSRANGKOON A,WORAMETRACHANON S,et al.Encapsulation of Lacto bacillus casei,01 by alginate along with himaize starch for exposure to a simulated gut model[J].Food Bioscience,2016,16:32-36.
[39]DIMITRELLOU D,KANDYLIS P,PETROVIC T,et al.Survival of spray dried microencapsulated Lactobacillus casei,ATCC 393 in simulated gastrointestinal conditions and fermented milk[J].LWT-Food Science and Technology,2016,71:169-174.
[40]LEONE R D S,ANDRADE E F D,ELLENDER-SEN L N,et al.Evaluation of dried yacon(Smallanthus sonchifolius)as an efficient probiotic carrier of Lactobacillus casei,LC-01[J].LWT-Food Science and Technology,2017,75:220-226.