酸菜来源植物乳杆菌体外去除胆固醇特性

对4株酸菜来源具有潜在益生特性的植物乳杆菌降胆固醇特性进行研究.研究发现其降胆固醇能力受胆盐浓度和胆盐种类的影响,当牛胆盐的添加浓度为3g/L时,降胆固醇能力达到最高(9.67mg/g～21.32mg/g[干重]);在培养基中添加不同种类的胆盐,降胆固醇能力不同,依次为牛磺胆酸钠＞混合胆盐＞甘氨胆酸钠.其产生的游离胆酸的量与胆固醇移除能力呈正相关,且以在牛磺胆酸钠存在下最佳.     

乳酸菌胆酸盐耐受性及降胆固醇能力研究

研究了分离自内蒙古传统稀奶油的四株乳酸菌的胆酸盐耐受性和降胆固醇能力。结果表明,供试菌株对牛磺胆酸钠的耐受性与对照菌株相比差异显著(P<0.05),其中屎肠球菌KLDS6.0330的耐受性最强;粪肠球菌KLDS6.0335呈现胆酸盐促进其生长的现象。菌株间胆固醇去除能力不同,在恒定pH5.5条件下去除能力更强(P<0.05);其中KLDS6.0330在不恒定pH和恒定pH5.5条件下分别达到58.47%和61.92%。四株供试菌株均具有共沉淀作用、吸收作用和降解作用。其中,干酪乳杆菌KLDS1.0344以共沉淀作用为主,KLDS6.0330以吸收作用和降解作用最为显著。     

产胆酸盐水解酶乳酸菌的鉴定及其生物信息分析

对初步筛选得到的胆酸盐水解酶活性高的2株乳杆菌进行了16SrDNA鉴定,结果表明,KLDS 1.0317和KLDS 1.0386均为植物乳杆菌。以牛磺脱氧胆酸盐和甘氨脱氧胆酸盐为底物,对两株植物乳杆菌所产BSH的比酶活进行了测定,试验证明了BSH具有底物特异性,且两株菌降解甘氨结合胆盐的能力强于牛磺结合胆盐;利用PCR技术克隆出胆酸盐水解酶基因,翻译成相应的氨基酸,并利用生物分析手段对bsh 1基因以及其预测氨基酸序列进行了对比分析,结果表明,两株植物乳杆菌的bsh 1基因非常保守,且预测的氨基酸序列的相似性非常高,不低于98%。     

具有胆固醇去除能力和胆盐水解酶活性的乳酸菌的体外筛选

对分离自内蒙古传统稀奶油或酸化稀奶油中的10株乳酸菌进行了胆固醇去除能力和胆盐水解酶活性的研究(定性和定量),筛选出了3株具有较高胆固醇去除能力和胆盐水解酶活力的乳酸菌菌株,分别为KLDS6.0330、KLDS6.0333和KLDS6.0335,并对它们进行16SrDNA分子生物学鉴定、酸耐受性和胆盐耐受性的测定。结果表明:这3株菌均为植物乳杆菌(L.Plantarum),并且表现出了较好的酸耐受性和胆盐耐受性。     

一株乳酸乳球菌的降胆固醇特性及其作用机制

探讨了乳酸乳球菌乳亚种HUCM 201的降胆固醇特性及其体外去除胆固醇的机制。乳酸乳球菌乳亚种HUCM 201菌株可从培养基中去除33.1%的胆固醇,其中14.3%的胆固醇发生共沉淀并重新溶解在洗涤液中,18.1%的胆固醇被吸收到菌体细胞内。HUCM 201菌株对5种结合型胆酸盐分别表现出了不同的胆盐水解酶活性,其中对甘氨胆酸钠的水解能力最强,总酶活为0.279 U/mL,比酶活为0.076 U/mg。     

植物乳杆菌C8的耐受性和体外降胆固醇活性研究

对一株分离自内蒙古奶豆腐中的植物乳杆菌C8进行了酸和胆汁盐耐受实验,抗生素敏感性试验以及体外胆固醇降解实验,并分析了其降胆固醇的作用机理.结果表明,C8能够耐受pH 3和高浓度胆盐(30g/L)环境;对所选10种抗生素均表现出一定的耐药性;该菌具有良好的降胆固醇活性,产生的游离胆酸的量与胆固醇移除能力呈正相关,且以在甘氨胆酸钠存在条件下最佳(5.39mmol/L).植物乳杆菌c8降胆固醇的作用机理主要是菌体细胞的表面吸附作用和胆固醇与胆盐的共沉淀作用.     

具有胆盐水解酶活力乳酸菌的筛选及16SrDNA分子生物学鉴定

对分离自内蒙古地区牧民家庭自制的2份酸马奶中的9株乳酸菌,进行胆盐水解酶活力的研究.采用定性和定量两种方法,筛选具有胆盐水解酶活力菌株,并对筛选出的茵株进行16S rDNA分子生物学鉴定.结果表明:9株菌中只有菌株18-1-3有白色颗粒状沉淀生成,其余8株均没产生.游离胆酸的生成量和牛磺胆酸钠的消失量分别为0.8524 mmol/L和0.8520 mmol/L.即游离胆酸钠的生成量和牛磺胆酸钠的消失量成比例.18-1-3菌株鉴定为发酵乳杆菌(Lb.fermentum).     

分离自Kefir的植物乳杆菌体外降胆固醇作用的研究

对分离Kefir中的植物乳杆菌体外降胆固醇作用进行了研究。结果表明，植物乳杆菌在含胆盐的培养基中具有良好的降胆固醇能力，24h后培养基中的胆固醇可降低60％(0．18g／L)；该菌体外降胆固醇作用的机理主要是其产生的胆盐水解酶水解胆盐变为游离态的胆酸，与胆固醇形成复合物，在酸性pH值条件下发生共沉淀。同时，也发现该菌产生的胆盐水解酶对反应底物(各种胆盐)的特异性不同，以脱氧牛磺胆酸最佳。     

胆盐水解酶基因在植物乳杆菌KLDS1.0386中的表达研究

本文从转录水平上研究了四种胆盐水解酶基因在植物乳杆菌KLDS1.0386中不同生长阶段的表达情况,并且分别以不同浓度的牛磺脱氧胆酸盐和甘氨脱氧胆酸盐为底物,以16s r RNA作为内参基因,利用实时荧光定量PCR技术研究不同浓度底物下的胆盐水解酶基因(bsh1、bsh2、bsh3、bsh4)表达的变化规律。结果显示,内参基因和目的基因的扩增效率均在90%~105%之间,说明目的基因的表达量可以被很好的反应出来。通过实时荧光定量PCR结果分析可知,植物乳杆菌KLDS1.0386中的4种胆盐水解酶基因随着生长时间的延长,表达量显著增加,而且在两种胆盐中都表达,基因表达量都被不同剂量(1%、2%、3%)的胆盐上调,上升幅度随着胆盐浓度的增加表现出极显著的增加趋势,而且胆盐不同,同浓度时每个基因的表达量也各不相同,在甘氨脱氧胆盐中的表达量普遍高于牛磺脱氧胆盐。     

植物乳杆菌耐酸耐胆盐的体外试验及其降胆固醇作用

目的:研究产胆盐水解酶植物乳杆菌H13的耐酸、耐胆盐特性,在此基础上对其可能的降胆固醇作用机理进行体外试验研究.方法:采用MRS培养基,模拟人体胃酸环境(pH=3)和胆盐环境(0.3%),测定H13菌株在酸性及高胆盐环境作用后的存活菌数,并在此胆盐浓度下测定植物乳杆菌H13的胆盐水解酶活力、降解胆盐能力和胆固醇沉淀量.结果:菌体能够耐受pH 3的环境和0.3%的胆盐环境;该菌产生的胆盐水解酶在降低胆固醇方面具有重要作用,酶活性与游离胆酸量、胆固醇沉淀量呈正相关,而且胆盐水解酶对反应底物的特异性不同,以甘氨胆酸钠为佳.结论:植物乳杆菌H13体外降胆固醇作用的机理主要是其产生的胆盐水解酶使胆盐失去共轭作用,在酸性(pH<6.0)条件下,解共轭胆盐与胆固醇形成复合物共同沉淀下来.     

Bile salt deconjugation and BSH activity of five bifidobacterial strains and their cholesterol co-precipitating properties

Five strains of bifidobacteria were screened for their bile salt deconjugation ability, bile salt hydrolase (BSH) activity and co-precipitation of cholesterol with deconjugated bile. Bile salt deconjugation was determined by the release of cholic acid. All strains exhibited deconjugation of both sodium glycocholate and sodium taurocholate. More cholic acid was liberated from the deconjugation of sodium glycocholate than sodium taurocholate. BSH activity was quantified by determining the amount of glycine or taurine liberated from conjugated bile salts by bifidobacteria strains. There was higher substrate specificity for glycine-conjugated bile compared to taurine-conjugated bile. Co-precipitation of cholesterol with cholic acid was observed from deconjugation of both sodium glycocholate and sodium taurocholate, and by all bifidobacteria strains studied. More cholesterol was precipitated with cholic acid when sodium glycocholate was used compared to sodium taurocholate. Increased cholesterol co-precipitation with deconjugated bile was observed with decreasing pH levels. Bifidobacterium infantis 17930 showed highest deconjugation ability and BSH activity towards bile mixtures that resemble the human bile, and may be a promising candidate to exert beneficial bile deconjugation activity in vivo.     

Bile salt deconjugation ability,bile salt hydrolase activity and cholesterol co-precipitation ability of lactobacilli strains

Eleven strains of lactobacilli were screened for their bile salt deconjugation ability, bile salt hydrolase activity (BSH) and co-precipitation of cholesterol with deconjugated bile. Bile salt deconjugation as determined by the release of cholic acid showed that more cholic acid was liberated from the deconjugation of sodium glycocholate than sodium taurocholate, and Lactobacillus acidophilus strains had higher deconjugation ability than L. casei strains. BSH activity, as quantified by the amount of taurine or glycine liberated from conjugated bile salts, indicated that substrate specificity was more towards glycine-conjugated bile compared to taurine-conjugated bile. Co-precipitation of cholesterol with cholic acid was observed from deconjugation of both conjugated bile, with more cholesterol being precipitated upon deconjugation of sodium glycocholate than upon that of sodium taurocholate. Cholesterol co-precipitation with deconjugated bile increased with decreasing pH. L. acidophilus ATCC 33200, 4356 and 4962 and L. casei ASCC 1521 showed highest deconjugation ability and BSH activity towards bile mixtures that resemble the human bile, and may be promising candidates to exert beneficial bile deconjugation activity in vivo.     

Bile salt hydrolase activity of three strains of Lactobacillus acidophilus

Three strains of Lactobacillus acidophilus, two from human intestinal origin (016 and L1) and one from porcine intestinal origin (ATCC 43121), were tested for their bile salt deconjugation activity. The L. acidophilus ATCC 43121 had more deconjugating activity of both sodium glycocholate and sodium taurocholate at pH 6.5 than did either L. acidophilus 016 or L1. The activity of intracellular bile salt hydrolase found in strain ATCC 43121 was 14-fold higher than that in either of the other two strains. The optimum pH for deconjugation of sodium glycocholate was between 4 and 5.5 for all three strains. For deconjugation of sodium taurocholate, the optimum pH was between 3.5 and 4.5 for strains L1 and ATCC 43121 and was between pH 5 and 6 for strain O16. The molecular mass of the enzyme in all three strains of L. acidophilus was estimated to be 126 kDa by Sephadex G-200 gel filtration. All three strains exhibited more bile salt hydrolase activity towards sodium glycocholate than towards sodium taurocholate.     

Binding of Bile Salts by Soluble Fibers and Its Effect on Deconjugation of Glycocholate by Lactobacillus acidophilus and Lactobacillus casei.

ABSTRACT: Binding of sodium cholate, sodium taurocholate, and sodium glycocholate by guar gum, soluble oat fiber, xanthan gum, and inulin was studied. All soluble fibers were able to bind sodium cholate, sodium taurocholate, and sodium glycocholate from a mixture of the three, but when tested individually, the fibers bound little or no sodium cholate. In general, the fibers bound higher concentrations of conjugated bile salts than they did free bile salts. Soluble oat fiber, which bound as much or more sodium cholate and sodium glycocholate as did the other 3 fibers, was tested for its effect on deconjugation of glycocholate by Lactobacillus acidophilus and Lactobacillus casei. The presence of the fiber significantly increased (P < 0.05) deconjugation by all cultures.     

Bile tolerance, taurocholate deconjugation, and binding of cholesterol by Lactobacillus gasseri strains

Bile tolerance, deconjugation of sodium taurocholate, and the cholesterol-binding ability of 28 strains of Lactobacillus gasseri were examined. There was significant variation among strains in growth in media containing bile and also variation in the ability to bind cholesterol. Cultures grown for 12 h at 37 degrees C bound significantly more cholesterol than did cells from a 48-h incubation. Variation among strains in the ability to deconjugate sodium taurocholate was not significantly different. Maximal deconjugation of sodium taurocholate was achieved with the cells during the stationary phase of growth (12 h). Statistical analysis showed no significant correlation between bile tolerance and sodium taurocholate deconjugation, bile tolerance and cholesterol-binding ability, or sodium taurocholate deconjugation and cholesterol-binding ability.     

Bile salt deconjugation activity of Propionibacterium strains and their cholesterol co‐precipitation abilities

The present study investigated the deconjugation of bile salts and co‐precipitation of cholesterol with deconjugated bile salts by seven Propionibacterium spp. strains in vitro. Propionibacterium spp. could deconjugate sodium glycocholate (1.53–5.31 mM) and sodium taurocholate (0.08–0.25 mM) bile salts. The highest cholesterol precipitation (47.8 µg/mL) was determined with Propionibacterium freudenreichii subsp. freudenreichii SP3 strain in a 0.15% oxgall‐containing medium. Significant (P < 0.05) correlations were observed between cholesterol co‐precipitation and deconjugation of sodium glycocholate among the strains. In vitro bile salt deconjugation activity studies of P. freudenreichii subsp. freudenreichii SP3 strain revealed that this strain may have potential as a probiotic strain for deconjugation of bile salts in vivo studies.     

Deconjugation and bile salts hydrolase activity by Bifidobacterium strains with acquired resistance to bile

Deconjugation by bile salts hydrolases in probiotics has been related to reduction of serum cholesterol levels in mammals. We compared the susceptibility to conjugated primary (glycocholate and taurocholate) and secondary (glycodeoxycholate and taurodeoxycholate) salts and the level of hydrolase activity of Bifidobacterium strains with acquired resistance to bile and of their more sensitive original strains. Minimum inhibitory concentrations against conjugated salts of the resistant strains were higher than that of the corresponding originals. None of the strains displayed deconjugation against primary salts, whereas most of them deconjugated secondary salts. Salts of cholic acid were more toxic than that of deoxycholic acid. Derivatives showed higher hydrolase activity than their originals. These results suggested a relationship between bile resistance and deconjugation. Finally, the resistance of bifidobacteria against glycodeoxycholate increased in the presence of maltose and cellobiose as compared with glucose, which could be related to a more efficient energy procurement from disaccharides.     

Mechanisms of cholesterol removal by lactobacilli under conditions that mimic the human gastrointestinal tract

Five strains of lactobacilli were studied for their ability to remove cholesterol in vitro under conditions that mimic the human gastrointestinal tract. The highest assimilation of cholesterol was observed in media supplemented with oxgall and the lowest in the presence of taurocholic acid. Scanning electron micrographs showed that cholesterol was adhered to the cellular surface of lactobacilli cells. Resting and dead cells were able to remove cholesterol although in small amounts. Additionally, inhibition of cholesterol micelles formation was observed in the presence of bile salts. All strains were able to deconjugate bile salts, where higher deconjugation was observed in the presence of sodium glycocholate compared with other bile salts studied. All strains also exhibited bile salt hydrolase activity and most strains showed higher substrate specificity towards glycine-conjugated bile than towards taurine-conjugated bile. The results indicated that lactobacilli could remove cholesterol in vitro via various mechanisms, and may exert such hypocholesterolaemic effects in vivo.     

Measurement of bile salt hydrolase activity from Lactobacillus acidophilus based on disappearance of conjugated bile salts

Bile salt hydrolase activity of Lactobacillus acidophilus was measured based on the disappearance of sodium glycocholate and sodium taurocholate from the reaction mixture using HPLC. The amount of sodium glycocholate and sodium taurocholate that disappeared was proportional to the amount of sodium cholate that appeared in the mixture as detected by HPLC. Sodium glycocholate did not precipitate at the enzyme reaction conditions (37 degrees C and pH 5.4) for determining bile salt hydrolase activity. The bile salt hydrolase assay was insensitive to low oxidation-reduction potential when measuring bile salt hydrolase from L. acidophilus, an intestinal microorganism. However, EDTA and freezing temperatures were necessary to maintain stability of the partially purified enzyme during storage.