具有降胆固醇功能乳酸菌的筛选及其生物学特性评价

目的 筛选具有降解胆固醇功能和良好生物学特性的乳酸菌。方法 以胆固醇为唯一碳源的基础盐培养基从健康人体粪便中筛选具有降解胆固醇功能的乳酸菌,采用16S rRNA基因测序分析确定菌株种属,对其耐酸、耐胆盐、耐过氧化氢、发酵上清液抑菌活性、肠上皮细胞黏附及胆固醇清除能力等生物学特性进行系统评价,并选用综合性能良好植物乳植杆菌进行动物实验,探究其体内的降胆固醇功能。结果 经16S rRNA基因测序分析,获得5株具有降胆固醇功能的植物乳植杆菌,对其生物学特性进行评测,发现5株菌均具有良好的耐酸耐胆盐耐过氧化氢能力,在pH 3.0的酸性条件下处理6 h,活菌数与初始菌量保持在同一数量级;3.0 g/L胆盐的条件下处理6 h,其活菌数保持在106 CFU/mL;1.0 mmol/L过氧化氢条件下处理6 h,活菌数显著增加。MRS发酵上清液对5种常见的食源性致病菌都有不同程度的拮抗作用,其抑菌圈直径均大于8.5 mm;具有良好的肠上皮黏附特性,对Caco-2细胞的黏附率均大于25%;胆固醇清除能力差异较大,其中植物乳植杆菌GLPL03和GLPL04的清除能力较强,达到59%和53%。综合菌株性能,选用植物乳植杆菌GLPL03和GLPL04进行为期12周的动物实验,发现植物乳植杆菌GLPL03和GLPL04能够显著降低小鼠血清总胆固醇和总甘油三脂水平,缓解小鼠高胆固醇血症的发展。结论 健康人体来源的5株植物乳植杆菌具有良好的生物学特性,可作为益生菌进行深入研究。     

产淀粉酶益生乳杆菌的筛选及鉴定

目的以健康仔猪肠道及粪便样品为基础,从中筛选产淀粉酶的乳杆菌菌株,并评价其作为益生菌候选菌株潜力。方法用MRS培养基分别从仔猪新鲜粪便和小肠黏膜上分离到乳酸菌,采用改良的产淀粉酶选择性培养基初筛得到能降解淀粉活性的菌株,并研究菌株的淀粉水解活性、抗逆能力、粘附特性及对抗生素的敏感性。结果从备选的485株乳酸菌中筛选得到具有初步淀粉酶活性的菌株25株（占总筛选数量的5.2%）,复筛选育得到具有较强淀粉酶活性的乳杆菌3株。进一步研究了这3株乳杆菌的抗逆能力、粘附特性以及对抗生素的耐药性,并对最终选育得到的菌株进行生理生化及16S rRNA分子鉴定。经选育鉴定的罗伊乳杆菌G8-5淀粉降解能力最强,并能耐受pH 3.0的酸度、1.0%的胆盐浓度,在小肠上皮细胞上的粘附效率超过15个以上,并对常用的抗生素具有较高的敏感率。结论罗伊乳杆菌G8-5符合安全益生菌的要求,可以作为产淀粉酶的益生乳杆菌优良的候选菌株。     

猪肠道乳杆菌的筛选及其生物学特性研究

目的从健康仔猪肠道中筛选到用于研制微生态制剂的优良乳杆菌株。方法本研究选用乳杆菌选择培养基LBS、MRS从健康仔猪肠道分离到67株乳杆菌,对其染色镜检、生化试验和耐酸、耐胆汁、耐高温、抑菌活性及动物安全性等生物学特性进行初步研究。结果从其中筛选到3株嗜酸乳杆菌,均能耐受pH3．0的酸度、0．8％～1．0％的牛胆盐和60℃的高温,其代谢产物对猪肠道致病性大肠埃希菌和猪沙门菌具有抑制作用,且对小白鼠无致病性。结论3株嗜酸乳杆菌符合益生菌的要求,可作为猪用益生素制剂的候选菌株。     

益生乳杆菌的筛选及特性研究

根据对优良益生菌的要求,从健康乳猪肠道中分离筛选到13株乳杆菌,其中嗜酸乳杆菌3株、干酪乳杆菌1株、链状乳杆菌1株、发酵乳杆菌8株。所选菌株能耐受1.0％的牛胆盐,对 pH3.0的酸度有较强的抵抗力,并能抑制常见致病菌,对小白鼠安全无毒,是制备益生素的优良菌种。     

乳酸菌的分离、鉴定及其生长特性

从动物粪便中分离乳酸菌,得到产酸快、代谢物抑菌活性强的乳酸菌菌株共６株。其中Ｏ－２菌株的产酸、抑菌、耐酸、耐胆盐和生长性能都优于其他菌株,是一株有潜质的有益菌菌性,经鉴定表明为乳杆菌。不同的营养配比的液体培养基进行发酵实验表明,Ｒ２培养基配料简单,成本低,发酵２４ｈ产生的菌体总数多,４８ｈ培养液的抑菌活性强,且其抑菌活性具有一定的热稳定性。     

一株降解生木薯淀粉的曲霉菌株的筛选、鉴定及其淀粉降解特性

本研究利用以生木薯淀粉为唯一碳源的筛选培养基,从腐烂木薯渣中分离筛选出一株可以降解生木薯淀粉的真菌菌株RSDF-7.根据RSDF-7形态和18S rDNA与28S rDNA之间的内转录间隔区(internal transcribed spacer,ITS)序列分析的结果,初步认定该菌株为曲霉属.菌株RSDF-7的粗酶液对多种不同的生淀粉底物均有水解效果;在以大米和玉米淀粉为底物时,其生淀粉分解活力比较高,分别为42%和40%.菌株RSDF-7的粗酶液具有良好的低pH稳定性,对生木薯淀粉的最适作用温度为50℃,最适作用pH为4.5.在30 min的吸附后,RSDF-7的粗酶液对生木薯淀粉的吸附力高达60%.使用HPLC对粗酶液的酶解产物进行检测,结果发现酶解产物中仅存在葡萄糖,表明菌株RSDF-7所产的生淀粉降解酶主要为糖化酶.扫描电镜观察结果发现,经RSDF-7粗酶液酶解后的生木薯粉颗粒破裂,形成空洞,说明RSDF-7粗酶液对生淀粉有较强的水解作用.可以预见,经纯化后的曲霉菌株RSDF-7生淀粉酶将来可以用于基于酶解的木薯淀粉转化.     

高对映选择性环氧化物水解酶产生菌的筛选及特性研究

从土壤中分离的芽孢杆菌Bacillus megaterium ECU1001所产五氧化物水解酶能高对映选择性水解缩水甘油苯基醚（对映选择率E值可达47.8）,当转化率为55.9%时,剩余的（S）－缩水甘油苯基醚的光学纯度（对映体过量值,ee）可达99.5%;当底物浓度提高到60mmol/L时,光学纯（S）－缩水基油苯基醚的收率达到25.6%。     

从实际应用特点出发筛选益生乳杆菌菌株

正众所周知,在人的体内和体表生活着大量的各类共生微生物。后者的总数量可达人体自身细胞数量的10-100倍,编码的基因数量可达人体自身的300多倍,被称为人体的\"第二基因组\",对保障人体的健康具[1]有重要意义。研发各种益生菌制剂来维持、调整、完善人体,特别是肠道的正常菌群结构,近年来一直[2]是备受关注的热点和前沿。乳杆菌作为公认的益生菌,可有效改善宿主的肠道环境、抑制有害菌的生长、     

豆豉中高产乳酸乳酸菌的筛选及其产酸条件的优化研究

目的 从云南传统发酵豆豉中分离并筛选得到一株高产乳酸的乳酸菌菌株,并对其产酸培养基组成及条件进行优化探讨.方法 运用紫外分光光度法,对豆豉由来高产有机酸菌株YM-4-3的碳源进行优化.结果 分子生物学鉴定结果表明该乳酸菌菌株属于植物乳杆菌,并将之命名为Lactobacillus plantarum YM-4-3,碳源优化结果表明,乳糖是L.plantarum YM-4-3 菌株生物合成乳酸的最佳碳源,而果糖则是最利于各种主要有机酸合成的碳源,由于此时各常见有机酸的含量均高于其他碳源时的含量,且其总酸含量高达9696.32 mg/L.结论 云南传统发酵豆豉可以成为功能性乳酸菌筛选的菌种资源库,所分离得到的L plantarum YM-4-3产酸能力较强,它可被应用于有机酸发酵及豆豉发酵的酸化处理.     

Genotypic analyses of lactobacilli with a range of tannase activities isolated from human feces and fermented foods

A total of 77 tannase producing lactobacilli strains isolated from human feces or fermented foods were examined for their genotypic profiles and intensities of tannase production. With a PCR-based assay targeting recA gene, all strains except one isolate were assigned to either Lactobacillus plantarum, L. paraplantarum, or L. pentosus whereas a 16/23S rDNA targeted PCR-based assay identified all except 6 isolates (inclusive of the above one isolate) as one of the closely related species. Subsequent DNA/DNA hybridization assays revealed that these 6 exceptional isolates showed low homology (between 1.2% and 55.8% relative DNA binding) against type strains of the three species. Supplemental carbohydrate fermentation profiles on the 6 isolates indicated that two of them were identified as L. acidophilus, one as Pediococcus acidilactici, one as P. pentosaceus, and two remained unidentifiable. The evidence suggests that the 16/23S rDNA targeted PCR assay can be used as a reliable identification tool for the closely related lactobacilli, and that the tannase gene is widely distributed within members of the Lactobacillaceae family. Meanwhile, a randomly amplified polymorphism DNA (RAPD) analysis revealed that all except 8 isolates were well allocated in 4 major RAPD clusters, though not species specific, consisting of two L. plantarum predominant clusters, one L. paraplantarum predominant, and one L. pentosus predominant. The RAPD patterns of the 8 non-clustered isolates, which consisted of the 6 unidentifiable isolates and 2 isolates identified as L. pentosus, were <40% similarity to those belonging to the 4 clusters. A quantitative assay of the tannase activities showed that there was a marked variation in the activities among the strains, which did not correlate with either species identification or clustering by RAPD.     

抗氧化菌株的筛选及其对HepG2细胞氧化应激损伤的保护作用

【背景】机体的氧化损伤会引发多种疾病和机体衰老,研究发现益生菌可以改善宿主的代谢水平,提高宿主的抗氧化能力,为抗氧化产品开发提供新方向。【目的】筛选具有良好抗氧化效果的菌株,为抗氧化药物以及保健品的开发和研究提供菌种材料。【方法】将从江西省宜春市温汤长寿村地区的健康人群粪便中筛选出的菌株进行16S rRNA基因鉴定,挑取其中6株益生菌,通过测定其培养液上清中的超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)酶活及1,1-二苯基-2-三硝基苯肼(1,1-dipheny-2-picrylhydrazyl,DPPH)自由基清除能力,选取一株抗氧化能力最佳的菌株进行后续细胞实验。建立HepG2细胞氧化应激损伤模型,使用CCK-8法评估其培养液上清对HepG2细胞氧化应激损伤的保护效果,通过试剂盒测定培养液上清对细胞中SOD酶活的保护作用,并进一步验证其培养液上清的抗氧化效果。【结果】筛选出的6株益生菌培养液上清中,菌株H0661表现出最佳的抗氧化能力,其培养液上清中GSH-Px酶活为586.11 U/mL (P<0.01),SOD酶活为1 278.00 U/mL (P<0.01),DPPH自由基的清除能力达到40.5%。结合16S rRNA基因鉴定结果,将其命名为木糖葡萄球菌(Staphylococcus xylosus) TG022。木糖葡萄球菌TG022培养液上清浓度为15%时对HepG2细胞氧化应激损伤的保护效果最佳(P<0.01)。此外,木糖葡萄球菌TG022培养液上清能有效保护HepG2细胞氧化应激损伤中SOD活性(P<0.01)。经木糖葡萄球菌TG022培养液上清处理后可以有效提高HepG2细胞的抗氧化活性(P<0.05)。【结论】本研究筛选出一株木糖葡萄球菌TG022,其培养液上清表现出良好的抗氧化效果,为研发抗氧化药物和保健品提供了一个潜在的菌株材料。     

Application of novel starter cultures for sourdough bread production

Sourdough application has been extensively increased in the last years due to the consumers demand for food consumption without the addition of chemical preservatives. Several starter cultures have been applied in sourdough bread making targeting the increase of bread self-life and the improvement of sensorial character. More specific, Lactobacillus acidophilus and Lactobacillus sakei as single and mixed cultures were used for sourdough bread making. Various sourdough breads were produced with the addition of sourdough perviously prepared with 10% w/w L. acidophilus, 10% w/w L. sakei and 5% w/w L. acidophilus and 5% w/w L. sakei at the same time. Various chemical parameters were determined such as lactic acid, total titratable acidity and pH. The results revealed that the produced sourdough bread made with sourdough containing the mixed culture was preserved for more days (12 days) than all the other breads produced in the frame of this study, since it contained lactic acid in higher concentrations. The respective total titratable acidity varied between 10.5 and 11 ml NaOH N/10. The same sourdough bread had a firmer texture, better aroma, flavor and overall quality compared to other sourdough breads examined in this study, as shown by sensory evaluation tests and results obtained through SPME GC–MS analysis, which revealed significant differences among the different bread types.     

泡菜水中产生物表面活性剂酵母菌株的筛选

[背景]由微生物产生的生物表面活性剂(biosurfactant,BS)具有低毒性、高效性、生物可降解性等多种特性,能在一定程度上缓解化学表面活性剂所造成的环境问题,因此筛选高产、安全的BS生产菌株备受研究者的关注.[目的]从泡菜水中筛选能代谢合成药食两用型BS的微生物菌株.[方法]运用滴崩法和排油圈法从传统发酵食品泡...     

In vivo Testing of Functional Properties of Three Selected Probiotic Strains

Summary Lactobacillus acidophilus M92, Lactobacillus plantarum L4 and Enterococcus faecium L3 were previously selected as probiotic strains on the base of in vitro selection criteria. To investigate functional properties of these three probiotic strains in vivo, Swiss albino mice were used as animal model. Survival, competition, adhesion and colonization were monitored in the gastrointestinal tract, as well as the immunomodulating capability of L. acidophilus M92, L. plantarum L4 and E. faecium L3. During the feeding of mice with probiotic strains with daily dose of 2 × 10¹⁰ rifampicin-resistant cells, the number of lactic acid bacteria in the faeces increased and reduction of enterobacteria and sulphite-reducing clostridia was observed. Rifampicin-resistant colonies of probiotic strains could be reisolated from the faeces of mice fed with the rifampicin-resistant cells. The similar results were obtained in homogenates of small and large intestine of mice on the first and fourteenth days after feeding with L. acidophilus M92, L. plantarum L4 and E. faecium L3. The adherence of the probiotic strains obtained in vitro correlated with their capability to adhere to mouse ileal epithelial cells in vivo. After oral immunization of mice with viable cells of L. acidophilus M92, L. plantarum L4 and E. faecium L3 with a daily dose of 2 × 10¹⁰ cells, the concentrations of serum IgA, IgG and IgM antibodies from all groups of mice were significantly higher in comparison to the control.     

Numerical phenetic study of the genus Carnobacterium

Eighty-nine strains representing the genus Carnobacterium, Enterococcus durans, Vagacoccus salmoninarum and atypical Lactobacillus strains MT12 and MT13 were examined for 92 unit characters. Computer analysis of the data resulted in the recovery of four major, five minor and thirteen single membered clusters. Three cluster-groups contained seventy-four of the Carnobacterium strains, Enterococcus durans NCFB 596^T and Lactobacillus maltaromicus NCFB 2382^T. Cluster-group A was equated with Carnobacterium piscicola and cluster-group B with Carnobacterium divergens. Lactobacillus maltaromicus NCFB 2382^T shared many properties in common with the C. piscicola strains. The recovery of several Carnobacteriumstrains as single membered clusters suggests that the genus Carnobacterium is underspeciated. Further work is also required to determine the subspecific structure of Carnobacterium divergens and Carnobacterium piscicola.     

Determining the Suitability of Lactobacilli Antifungal Metabolites for Inhibiting Mould Growth

Summary In recent years, public concern about indoor mould growth has increased dramatically in the United States. In this study, lactic acid bacteria (LAB), which are known to produce antimicrobial compounds important in the biopreservation of food, were evaluated to determine if the same antimicrobial properties can be used to inhibit mould fungi that typically colonize wood. Based on biomass measurement, cell-free supernatants from Lactobacillus casei subsp. rhamnosus and Lactobacillus acidophilus grown in deMan Rogosa Sharpe (MRS) broth inhibited 95–100% growth of three mould fungi and one stain fungus associated with wood-based building materials. Lactic acid and four unknown compounds ⩽ kDa molecular weight were fractionated from the culture supernatant by thin layer chromatography and high-performance liquid chromatography. Antifungal activity, which was attributed to one or more unknown metabolites, was retained during heating and neutralization. A 1:2 dilution of L. casei supernatant inhibited 100% growth of all test fungi.