牦牛曲拉源乳酸菌的耐受性与抑菌性能比较

以源于牦牛曲拉的6株益生乳酸菌为研究对象,比较了耐酸能力、耐胆盐能力、耐渗透压能力和抑菌性能。结果表明:6株乳酸菌菌株的耐酸能力、耐胆盐能力、耐渗透压能力和抑菌性能各不相同。菌株G2、G4、Q1对酸性环境的耐受性最好,G2和G4的耐胆酸盐的能力最强,G2、G4、Q1对高渗透压的耐受性较强,所有菌株在模拟胃肠液环境下的存活率均在90%以上;菌株Q2对大肠杆菌的抑菌效果最强,菌株Q1次之;菌株Q1和Q2对金黄色葡萄球菌的抑制效果较好,其次是菌株G2;对沙门氏菌抑菌作用最强的为菌株Q2,抑菌能力最差为G3;对单增李斯特菌抑菌作用最强的为菌株G4。研究结果可为牦牛曲拉源益生乳酸菌的应用提供理论依据。     

甘南州牧区优良乳酸菌耐受特性的研究

对分离自甘南州牧区酸牦牛奶中的4株优良乳酸菌的生长性能、耐酸能力、耐胆盐能力、耐渗透压能力和抑菌能力进行了研究。结果表明:4株乳酸菌均有较好的生长特性;球菌A-2-1和杆菌F-2-1在极限pH1.5仍能生长;当胆酸盐浓度达到0.5%时,杆菌B-4-2-1生长受到抑制,其余3株菌均能生长;杆菌F-2-1和G-3-2对浓度高达8%的NaCl均表现出一定的耐受力;同时4株乳酸菌也表现出良好的抑菌效果,杆菌B-4-2-1对金黄色葡萄球菌抑制作用较强,抑菌圈直径19mm,在对大肠杆菌和枯草芽孢杆菌的抑制作用中菌株F-2-1效果明显,抑菌圈直径分别为18mm和22mm,研究结果可以为其在发酵乳制品中的应用提供参考依据。     

抑菌活性乳酸菌的筛选及其抑菌物质的提取条件优化

从内蒙古东部地区发酵酸菜中分离的乳酸菌为供试菌株,以大肠杆菌和金黄色葡萄球菌为指示菌进行了抑菌活性筛选,并检测其抑菌谱。通过单因素和L_9(3~4)正交实验确定抑菌活性物质的最佳提取条件,并应用于市售牛饲料中检测其抑菌效果。结果表明,在发酵酸菜中分离的14株供试乳酸菌中获得了1株抑菌活性较强且稳定的菌株S1-4,该菌株对大肠杆菌和金黄色葡萄球菌的抑菌圈直径分别达(19.69±0.71)mm和(20.59±0.26)mm,并呈现出较广的抑菌谱。菌株S1-4产抑菌活性物质的最佳提取条件:沉淀p H为5.0,乙醇浓度95%,沉淀时间8 h,料液比为1∶4(v∶v),在此条件下,抑菌圈直径为39.53 mm,提高了23.18%。菌株S1-4发酵液添加到牛饲料中,可抑制饲料中细菌和霉菌的生长,并且随添加量的增加,抑菌效果明显增加。     

两株乳酸菌的耐受特性分析与研究

对两株乳酸菌植物乳杆菌A（L.plantarum）以及短乳杆菌B（L.brevis）的耐渗透压、耐胆酸盐、耐酸能力和抑菌能力等耐受特性进行了分析与研究。通过使用分光光度计来测定菌悬液的OD值以及含乳酸菌的纸片抑制大肠杆菌（E.coil）和金黄色葡萄球菌（S.aureus）实验来观察2种菌株的耐受特性。实验结果表明：两种菌株都有一定的耐受特性能力,在相同的环境条件下,每个菌株都显现出不同的抗性能力,各有着自己对某种条件的耐受能力优势。通过耐受特性和抑菌特性综合分析可以得出, 菌株A是比菌株B更具有益生潜力的优良菌株。     

抗龋齿口腔益生乳酸菌的筛选

本研究旨在筛选出抑制口腔致病菌能力强、安全性高、口腔定植能力强的乳酸菌。首先,采用牛津杯琼脂扩散法从实验室乳酸菌菌种资源库中筛选抑制变异链球菌和戈氏链球菌的菌株,对抑菌性较高的28株乳酸菌菌株进行了溶血性评价,结果显示7株菌株呈现α-溶血,21株菌株无溶血性。通过中和处理、过氧化氢酶以及蛋白酶敏感性研究,探究无溶血性且对变异链球菌和戈氏链球菌抑菌圈直径≥10 mm的16株乳酸菌的抑菌代谢产物,筛选出7株抑菌能力强并有可能产生非有机酸类抑菌代谢产物的乳酸菌。安全性评价结果显示：7株乳酸菌产酸性以及对牙齿腐蚀性均较低,除菌株JZ14-1对溶菌酶最高耐受质量浓度为1.6 mg/mL,其他6株乳酸菌对溶菌酶最高耐受质量浓度为1.2 mg/mL,均可耐受溶菌酶。菌株HB13-2敏感的抗生素数量最多。口腔定植能力评价显示：菌株HB13-2具有最高的表面疏水性为44.00%,表面酸电荷为20.16%,表面碱电荷为49.44%,胞外多糖质量浓度为0.283 mg/mL;菌株HB13-2具有较低的自身生物膜形成能力和良好的自聚能力,易定植于口腔。菌株HB13-2对变异链球菌生物膜最高抑制率为59.72%...     

肉源乳酸菌抑菌特性分析

为了筛选有抑菌活性的乳酸菌并分析其抑菌的物质基础,利用牛津杯琼脂扩散法,分析了19株从牧区风干肉制品中分离得到的肉源乳酸菌发酵上清液对大肠杆菌和金黄色葡萄球菌的抑菌能力,同时从中分别选取对大肠杆菌和金黄色葡萄球菌抑制能力有显著差异的乳酸菌各3株,对其上清液中的抑菌物质基础和抑菌能力影响因素进行了分析。结果表明19株乳酸菌中菌株F19对大肠杆菌的抑菌活性最强,抑菌圈直径为(15.07±0.55) mm,而F11发酵上清液对大肠杆菌完全没有抑制效果,菌株F16对金黄色葡萄球菌的抑菌活性最强,抑菌圈直径为(14.47±0.38) mm,而F6、F2发酵上清液对金黄色葡萄球菌完全没有抑制效果;抑菌的物质基础分析表明,酸性环境在抑菌过程中占主导地位,细菌素与过氧化氢发挥协同作用;对其发酵上清液进行不同pH以及不同温度处理后发现,其抑菌活性在pH<5.0时随pH的降低而显著增强(P<0.05),当pH分别升高至5.0、6.0、7.0时试验乳酸菌发酵上清液均不具有抑菌能力,F18发酵上清液分别在60、80、100、121 ℃热处理一定时间后其抑菌能力与对照组相比仍然无明显差异(P>0.05),F11发酵上清液在100 ℃热处理30 min后抑菌能力显著减弱(P<0.05),其余实验菌株发酵上清液温度达到60 ℃以上,抑菌能力随热处理温度提高而显著(P<0.05)下降。本研究发现了乳酸菌发酵上清液的抑菌效果具有特异性,抑菌物质主要是酸性物质,其次是细菌素和过氧化氢;抑菌能力受pH和温度影响,同时具有特异性。研究结果可为优质菌种资源开发提供理论指导与技术支持。     

南山牧场牛乳中具有抑菌活性乳酸菌的筛选及鉴定

该研究采用湖南南山高山牧场的鲜牛奶为原料,李斯特菌为指示菌,抑菌圈直径为评价指标,从中筛选抑菌特性较高的乳酸 菌,通过形态观察和分子生物技术对其进行鉴定。 同时,对菌株所产的抑菌物质的成分进行初步分析,并研究pH、温度及NaCl对其抑 菌效果的影响。 结果表明,从湖南南山高山牧场的鲜牛奶中共分离出28株乳酸菌,其中7株对李斯特菌具有抑菌活性,且菌株C15的 抑菌能力最强,抑菌圈直径为6.17 mm。 菌株C15被鉴定为乳酸乳球菌（Lactococcal lactis）,初步判定其所产的抑菌物质成分为蛋白类 或多肽类物质,该抑菌物质在酸性及中性条件下稳定,且具有较好的热稳定性和耐盐性,经80 ℃处理20 min后,抑菌活性保留54.35%; 经10%NaC（l 1 mol/L）处理2 h后,抑菌活性保留86.76%。     

抑制阴道加德纳菌乳酸菌的筛选及安全性研究

为筛选具有较强抑制阴道加德纳菌(Gardnerella vaginalis)特性的乳酸菌菌株，该研究通过双层琼脂扩散法测定菌株抑制阴道加德纳菌的能力，从筛菌样品中筛选能力较强的乳酸菌，通过溶血性试验、动物致病性试验评价其安全性，并进行了耐酸耐胆盐能力试验。结果表明，筛选菌株中罗伊氏粘液乳杆菌(Limosilactobacillus reuteri)HCS02-001抑菌能力最强，发酵上清液抑菌圈直径为18.23 mm，发酵上清液经过3倍浓缩后抑菌圈直径可达21.58 mm。菌株HCS02-001无溶血性，无致病性，在pH 2.0条件下培养17 h活菌数对数值为7.97，存活率为85.52%，在1.5%胆盐条件下培养17 h活菌数对数值为8.48，存活率为90.99%。结果显示罗伊氏粘液乳杆菌HCS02-001具有较强的抑制阴道加德纳菌特性，初步判定其具备安全性。     

驼乳制品中抑制α-淀粉酶和α-葡萄糖苷酶活性乳酸菌的筛选及益生特性研究

目的:本研究以新疆阿勒泰地区的驼乳制品为研究对象,筛选对α-淀粉酶和α-葡萄糖苷酶有抑制活性的优良乳酸菌。方法:采用稀释涂布法分离纯化菌株,DNS和pNPG法筛选对α-淀粉酶和α-葡萄糖苷酶有抑制活性的菌株,通过耐酸性、耐胆盐、模拟胃肠道环境耐受性、抑菌性、抗氧化活性实验评价菌株的益生特性。结果:从驼乳制品中共计获得34株菌株,其中6株对α-淀粉酶和α-葡萄糖苷酶有抑制活性,经过形态学和16S rRNA分子鉴定,确定有4株为植物乳杆菌Lactiplantibacillus plantarum,2株为副干酪乳杆菌Lactiplantibacillus paracasei。6株乳酸菌对α-淀粉酶的抑制活性都达到50%以上,其中X34对α-淀粉酶的抑制率最高达到88.59%。本实验中筛选的乳酸菌对α-葡萄糖苷酶的抑制率在11%~16%之间,X31抑制率最高为15.43%。6株乳酸菌在不同pH（1.0、2.0、3.0）和不同浓度胆盐（1、2、3 g/L）的培养基中均可存活。6株乳酸菌在模拟胃液和肠液中的存活率分别达到83%和90%以上。对羟自由基和超氧阴离子自由基的清除率超过90%以上,其中X29对羟自由基的清除率最高为92.43%,对超氧阴离子自由基清除率最强的是X33清除率达到94.04%。6株乳酸菌对大肠杆菌、沙门氏菌和金黄色葡萄球菌有一定的抑制作用,其中X31对大肠杆菌的最大抑菌圈直径为19.63 mm,X23对沙门氏菌抑菌圈最大达到19.85 mm,菌株X33对金黄色葡萄球菌的抑菌圈直径最大为19.17 mm。结论:从驼乳制品中筛选到6株具有潜在降糖活性的乳酸菌,对强酸、胆盐和胃肠液有一定的耐受性,抗氧化能力较强,具有抑制致病菌的效果,为后期研发功能性降糖饮品提供益生菌株。     

云南建水酸浆豆腐中乳酸菌的生长特性

为了筛选出优良的乳酸菌进行发酵制作酸浆,对分离自云南建水豆腐酸浆中的五株乳酸菌(SYG01、SYG02、SYG03、SYG04、SYG05)的生长曲线、产酸能力、耐酸能力和耐渗透压能力进行了比较。生长曲线实验表明在相同培养时间内,菌株SYG02繁殖能力最强,其次是菌株SYG03,而菌株SYG05、SYG04、SYG01的生长速率明显较慢;在产酸方面SYG02产酸能力最强,速度最快,SYG03、SYG05和SYG04次之,SYG01最弱。在耐酸方面,在pH4.0的环境下5株菌生长良好,在pH3.0的酸性环境下5株菌虽然能够存活,但活菌数的数量级仅在102~104 CFU/mL,其中菌株SYG02和SYG03比其它三株菌株表现出较好的耐酸能力。在耐渗透压方面,SYG02在8%(w/V)的NaCl中依然表现出较强的耐受性,活菌数的数量级达到107 CFU/mL,其余4株菌在NaCl含量大于6%(w/V)时生长受到明显抑制。并以发酵黄浆水的pH及产酸量为指标对菌株的产酸能力进行比较,发现混合菌株的产酸能力高于单菌株,其中菌株SYG02和菌株SYG03组合发酵的效果最好,在发酵72 h后黄浆水的pH为3.52,产酸量达到6.46 g/L。从而得出菌株SYG02具有良好的生长、产酸和耐渗透压能力,具有良好的应用潜力,对酸浆豆腐的工业化生产有重要的意义。     

Functional Properties of Lactic Acid Bacteria Isolated from Romanian Fermented Vegetables

A total of 139 lactic acid bacterium (LAB) strains isolated from Romanian traditionally fermented vegetables were screened for the ability to produce exopolysaccharides and for their antagonistic activity against a set of nine LAB strains, three Bacillus strains, and four Gram-negative bacteria. Eighty-five of the tested strains showed a variable antimicrobial activity against Listeria monocytogenes ATCC 1911, 35 of the strains showed a limited inhibition zone against Escherichia coli ATCC25922, and 26 strains against Salmonella enterica ATCC 14024, while 19 strains showed inhibition against one or all three Bacillus strains used as indicators. None of the tested strains showed an antimicrobial activity against Staphylococcus aureus ATCC 25923. Several strains showed antibacterial activity against more than one indicator strain. For instance, Lactobacillus plantarum 307, Lactobacillus brevis 308, and Lactobacillus plantarum/pentosus 358 were active against five of the indicator strains used, while other 23 LAB were active against three indicator strains. In the case of two strains, namely Leuconostoc citreum 344 and Lactobacillus brevis 183, the activity was maintained after neutralizing the pH of the cell-free supernatant likely due to the production of bacteriocins. The gel permeation chromatography-based screening revealed seven EPS-producing LAB strains. Two of the positive strains, namely Leuconostoc citreum 177 and Leuconostoc citreum 52, have been shown to produce large amounts of EPS, of about 20 g/L. All isolated EPS have a high molecular mass, of above 1400 KDa, and a monomer composition dominated by the presence of glucose.     

酵素中具有抑菌能力乳酸菌的筛选及鉴定

该研究探讨了从酵素中分离纯化具有抗菌性能的12株乳酸菌，对其进行生理生化实验和API鉴定。利用牛津杯法对阪崎大肠杆菌、鼠伤寒沙门氏菌和宋内志贺氏菌、大肠杆菌、金黄色葡萄球菌进行抑菌特性研究，并探讨了乳酸菌是否产生物胺能力和偶氮还原酶。结果表明，酵素源的12株乳酸菌对肠道致病菌均具有不同程度的抑菌活性，其抑菌圈直径为14~22 mm，对大肠杆菌的抑菌效果较好，其抑菌圈直径为15~23 mm，对金黄色葡萄球菌的抑菌圈直径为14~24 mm，12株乳酸菌都不具有产生物胺和偶氮还原酶的能力。通过生理和API生化鉴定确定XZJ003、XZJ006、XZJ015均为副干酪乳杆菌，通过16S rDNA和同源性确证XZJ015为副干酪乳杆菌，该研究结果对今后益生菌的深入研究和功能性食品的开发具有一定的作用。     

Invited review: methods for the screening, isolation, and characterization of exopolysaccharides produced by lactic acid bacteria

The ability to produce exopolysaccharides (EPS) is widespread among lactic acid bacteria (LAB), although the physiological role of these molecules has not been clearly established yet. Some EPS confer on LAB a "ropy" character that can be detected in cultures that form long strands when extended with an inoculation loop. When EPS are produced in situ during milk fermentation they can act as natural biothickeners, giving the product a suitable consistency, improving viscosity, and reducing syneresis. In addition, some of these EPS may have beneficial effects on human health. The increasing demand by consumers of novel dairy products requires a better understanding of the effect of EPS on existing products and, at the same time, the search for new EPS-producing strains with desirable properties. The use of genetically modified organisms capable of producing high levels of EPS or newly designed biopolymers is still very limited. Therefore, exploration of the biodiversity of wild LAB strains from natural ecological environments is currently the most suitable approach to search for the desired EPS-phenotype. The screening of ropy strains and the isolation and characterization of EPS responsible for this characteristic have led to the application over the past years of a wide variety of techniques. This review summarizes the available information on methods and procedures used for research on this topic. The information provided deals with methods for screening of EPS-producing LAB, detection of the ropy phenotype, and the physicochemical and structural characterization of these molecules, including parameters related to their viscosifying properties. To our knowledge, this is the first compilation of methods available for the study of EPS produced by LAB.     

Assessment of potential probiotic properties of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains isolated from kefir

In this study, the metabolic activities (in terms of quantities of the produced lactic acid, hydrogen peroxide, and exopolysaccharides) of 8 strains of Lactobacillus spp., Lactococcus spp., and Pediococcus spp., were determined. Lactic acid levels produced by strains were 8.1 to 17.4 mg/L. The L. acidophilus Z1L strain produced the maximum amount (3.18 μg/mL) of hydrogen peroxide. The exopolysaccharides (EPS) production by the strains was ranged between 173 and 378 mg/L. The susceptibility of 7 different antibiotics against these strains was also tested. All strains were found to be sensitive to ampicillin. The tolerance of the strains to low pH, their resistance to bile salts of strains, and their abilities to autoaggregate and coaggregate with Escherichia coli ATCC 11229 were also evaluated. High EPS-producing strains showed significant autoaggregation and coaggregation ability with test bacteria (P < 0.01). A correlation also was determined between EPS production and acid-bile tolerance (P < 0.05). EPS production possibly affects or is involved in acid-bile tolerance and aggregation of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains and supports the potential of L. acidophilus Z1L strain as new probiotic.     

产胞外多糖植物乳杆菌的筛选及粗多糖的活性研究

通过观察乳酸菌菌落拉丝状况并测定其胞外多糖（exopolysaccharide,EPS）的产量,筛选出1 株所产EPS黏性好、产量高的乳酸菌AR307,经鉴定为植物乳杆菌。为得到更多的胞外多糖,对植物乳杆菌AR307的发酵条件进行优化,确定其在发酵温度32 ℃、发酵时间20 h条件下的产糖量为389 mg/L。在体外实验中,所得胞外多糖具有抑制HT-29肿瘤细胞活性、降低血糖水平的作用。     

药食两用植物提取液对李果实采后常见致腐真菌抑制作用

采用滤纸片法比较七种植物提取液对李果实采后常见致腐真菌的抑菌效果,研究甘草和肉桂复合提取液对褐腐病菌（Monilinia fructicola）的抑制效果及抑菌机理。结果表明,1.0g/mL甘草提取液对褐腐病菌抑制作用最强,抑菌圈直径为12.03mm;1.0 g/mL肉桂提取液对酵母菌（Saccharomyces）、黑曲霉（Aspergillus niger）、米曲霉（Aspergillus oryzae）抑制作用最强,抑菌圈直径分 别为21.25 mm、17.49 mm和13.28 mm。 甘草、肉桂提取液对褐腐病菌的最小抑菌浓度（MIC）值分别为7.81 mg/mL、15.63 mg/mL,两者 联合使用时对褐腐病菌的分级抑菌浓度（FIC）值为0.75,抑菌效果增强。 甘草、肉桂提取液处理后的褐腐病菌培养液相对电导率分别 达到96.64%、48.47%;可溶性蛋白质含量分别达到50.6 μg/mL、25.1 μg/mL。 推测其破坏了褐腐病菌细胞膜结构,改变了细胞膜渗透性,导致细胞质外泄,从而发挥抑菌作用。     

Factors influencing autoaggregation and aggregation of Lactobacillus delbrueckii subsp. bulgaricus isolated from handmade yogurt

Of 26 Lactobacillus delbrueckii subsp. bulgaricus strains isolated from yogurt, strains B2 and 22, which produce low levels (28 and 21 mg liter(-1), respectively) of extracellular polysaccharides (EPSs), and strains B3 and G12, which produce high EPS levels (211 and 175 mg liter(-1), respectively), were selected for further study. The two high EPS-producing strains showed a significant autoaggregation and coaggregation ability with Escherichia coli ATCC 11230 (P < 0.05). Moreover, the effect of bile was evaluated on autoaggregation and hydrophobicity. Autoaggregation and hydrophobicity of these L. delbrueckii subsp. bulgaricus strains decreased after treatment with bile. Only the high EPS-producing L. delbrueckii subsp. bulgaricus strain B3 showed greater autoaggregation (80%) and hydrophobicity (86%) than the other strains after bile treatment. When these strains were assessed for the inhibition of E. coli ATCC 11230 in coculture, L. delbrueckii subsp. bulgaricus B3 completely inhibited E. coli during 24 and 48 h of incubation. This investigation showed that a high EPS production and coaggregation ability may be important in the selection of probiotic strains.