芒果源植物乳杆菌FMNP01的全基因组测序及序列分析

目的:植物乳杆菌FMNP01是从新鲜热带芒果中分离获得的一株具有较好抗菌活性的乳酸菌。为深入研究其内在抗菌机理,对其进行全基因组测序并解析基因组序列信息。方法:采用高通量测序技术对植物乳杆菌FMNP01进行全基因组测序,采用生物信息学方法对其进行序列组装、基因预测与功能注释,并深入挖掘其细菌素合成相关基因簇信息。结果:通过基因组组装共得到18个重叠群,整个基因组大小为3 313 644 bp,GC含量44.48%。将序列提交至Gen Bank数据库,登陆号为JPSU00000000。在其基因组中发现一段细菌素合成相关基因簇。结论:本研究结果揭示了植物乳杆菌FMNP01抑菌机理,为其功能基因组学研究提供了理论依据。     

干酪乳杆菌Zhang分类学地位及潜在益生相关基因分析

干酪乳杆菌Zhang是本研究团队筛选出的具有优良特性的益生乳酸菌菌株。传统方法难以区分干酪乳杆菌及其近缘物种。因最近乳杆菌的系统分类进行了更新，故导致Zhang的分类地位需重新确认。本研究利用比较基因组学方法分析Zhang的分类学地位、耐药基因、致病性基因和环境抗性基因特征，探究该菌株基因组上携带的细菌素基因簇和潜在益生基因。比较基因组学结果显示Zhang与副干酪乳酪杆菌模式菌株ATCC 25302T的平均核苷酸一致性值是98.49%，通过分类学数据库比对和系统发育分析确认Zhang的分类学地位为副干酪乳酪杆菌。Zhang的基因组不含有耐药基因、致病性基因和环境抗性基因。在其基因组上注释到核心肽、LanT转运和引导切割及免疫或运输功能等Carnocin细菌素基因簇。此外，Zhang的基因组编码了谷胱甘肽合成、核黄素合成、黏附因子和分泌胞外多糖等潜在益生基因。本研究更新了Zhang的分类学地位为副干酪乳酪杆菌，揭示其不含有耐药基因、致病性基因及环境抗性基因，并注释到多个潜在益生基因，为该菌株进一步开发及产业化提供了数据参考。     

不同来源的植物乳杆菌基因组和抗生素抗性

为了研究不同分离源对植物乳杆菌基因组和功能的影响,选取了来自发酵酱、泡菜、粪便3种环境的33株植物乳杆菌,通过比较基因组学手段研究菌株的基因组基本特征、直系同源基因、系统进化关系,并结合功能基因注释结果与表型结果分析菌株对抗生素的耐药性。系统发育树揭示了分离源对植物乳杆菌的遗传进化具有较为显著的影响;同源基因结果表明粪便源的菌株其特殊基因数量要高于泡菜源和发酵酱源菌株的数量。33株植物乳杆菌均含有环丙沙星、四环素、氯霉素、甲氧苄氨嘧啶和万古霉素的抗性基因,菌株对这些抗生素也表现出抗性;绝大多数菌株对庆大霉素、链霉素、卡那霉素、新霉素、氨苄西林敏感,在基因组中也没有相关抗性基因;而红霉素和克林霉素的基因和表型并不对应。抗生素实验结果说明,分离源对菌株影响较小,大部分基因型与表型可以对应,基因组学对研究植物乳杆菌的生理特性起一定的指导作用。     

长双歧杆菌W13 的全基因组序列分析

长双歧杆菌由于其具有的益生功能而备受关注，该文旨在基于全基因组分析，揭示长双歧杆菌W13（Bifidobacterium longum W13）的基因特性，分析其功能基因和代谢通路。结果表明，长双歧杆菌W13 隶属于长亚种，通过直系同源蛋白分组比对（evolutionary genealogy of genes：non-supervised orthologous groups，eggNOG）数据库注释，发现氨基酸代谢、碳水化合物代谢是该菌株的主要代谢功能；通过京都基因和基因组百科全书（kyoto encyclopedia of genes and genomes，KEGG）数据库注释，发现菌株中含有较多的群体感应相关基因，表明该菌株可促进肠道稳态。通过碳水化合物活性酶（carbohydrate-active enzymes，CAZy）数据库注释，发现该菌株具有较高的糖合成能力。最后，通过比较基因组分析，发现长双歧杆菌长亚种的遗传多样性和稳定性较高，相较于其他长双歧杆菌长亚种，菌株W13 含有其特定的功能基因。综上表明菌株W13 具有一定的益生功能，有开发为益生菌的潜力。     

母乳婴儿源植物乳杆菌BF_15体外抗氧化活性及缓解DSS诱导小鼠结肠氧化损伤的研究

以鼠李糖乳杆菌GG(LGG)为对照菌株,研究了自行分离鉴定的母乳婴儿源益生菌植物乳杆菌BF_15的体外抗氧化活性、安全性与缓解DSS诱导小鼠结肠氧化损伤的症状.结果表明:试验菌株植物乳杆菌BF_15能够耐受高浓度的H2O2(3.5 mmol/L).发酵上清液中,菌株BF_15对DPPH的清除能力低于对照菌株LGG(P<...     

双歧杆菌、乳杆菌和芽孢杆菌产细菌素和多糖研究进展

双歧杆菌、乳杆菌和芽孢杆菌是益生菌的重要成员,具有很大的开发潜力。其不仅能作为优势菌调节肠道微生态平衡、防治疾病,还能产生具有广谱抗菌活性的细菌素和多糖类等化合物,其中有些化合物还具有抗肿瘤和抗氧化等特性,这些特性使得这些化合物和菌株本身有望开发成为新型膳食补充剂和保健功能性食品。但许多化合物的结构、活性、机制以及安全性尚不明确。因此,本文总结了2010~2019具有益生潜力的双歧杆菌属、乳杆菌属和芽孢杆菌属细菌中分离得到的细菌素和多糖化合物,并对其分类、结构、抗菌、抗肿瘤和抗氧化等功能进行综述,为菌株及其代谢产物的开发和应用提供参考。     

风干肉中产脂肪酶瑞士乳杆菌TR13全基因组测序及序列分析

瑞士乳杆菌TR13是从自然风干羊肉中分离筛选到的1株产脂肪酶活性高的菌株,为能够更好地研究该菌株在羊肉发酵过程中的代谢机理和功能,采用PacBio Illumina HiSeq测序平台对细菌TR13进行完成图测序分析。结果表明,瑞士乳杆菌TR13基因组为1套环状无质粒分子,基因组序列总长度为2 172 224 bp,GC含量为36.82%。基因组中预测到2 536个编码基因,编码基因总长度为1 883 532 bp,平均长度为799.46 bp,平均密度为1.08个/kb,对应蛋白质平均序列长度为265 aa,占基因组百分比为86.71%。TR13菌株基因组中包含15个rRNA操纵子和63个tRNA。预测到可能的毒力基因75个,耐药基因150个。编码基因通过GO、COG和KEGG数据库的对比分析,完成了TR13菌株基因组基本功能注释和代谢通路基因信息注释,注释信息可为菌株应用研究提供一定理论依据。     

母乳及婴儿粪便菌群分析及功能菌株筛选

目的 探究母乳喂养婴儿粪便菌群及母乳菌群相关性,获得具有潜在益生功能菌株.方法 对健康母子母乳及婴儿粪便样本进行Illumina高通量测序,探究母乳与婴儿粪便样本菌群结构组成,母乳菌群在婴儿肠道垂直传递情况.通过体外益生性质研究,对分离的菌株进行筛选.结果 母乳中的主要菌科为肠杆菌科、莫拉氏菌科、葡萄球菌科和链球菌科;...     

植物乳杆菌KLDS1.0391在酸奶体系中的细菌素产生特点

植物乳杆菌KLDS1.0391能够合成细菌素,也是益生菌,在食品中既可作为益生菌使用,也可作为辅助发酵剂用于生物防控,该研究主要考察了KLDS1.0391菌株在酸奶体系中细菌素的产生特点。研究结果表明,在发酵的6 h期间,抑菌活性随发酵时间延长而增强,发酵结束时,添加植物乳杆菌KLDS1.0391酸奶组的抑菌活性显著高于仅使用酸奶发酵剂的对照组(P<0.01)。与对照组相比,加入辅助发酵剂的实验组的感官品质未发生明显的变化。植物乳杆菌KLDS1.0391具备开发益生酸奶的潜力。     

新疆地区母乳中乳酸菌的分离鉴定及潜在益生菌的筛选

母乳是婴儿营养物质的主要来源且蕴藏着丰富的乳酸菌资源。为了了解新疆地区母乳中乳酸菌组成,筛选母乳中具有优良益生特性的双歧杆菌。该文通过传统纯培养方法对6份新疆地区母乳中的乳酸菌进行分离鉴定。以益生菌株Bifidobacterium lactis BB12作为对照,通过发酵特性、体外胃肠液耐受性及胆盐耐受性试验对双歧杆菌进行筛选,得到2株具有潜在益生特性的双歧杆菌,对其进行最适pH,最适温度,对NaCl的耐受性,生长曲线及菌株产酸能力等生理生化特性试验。从6份样品中共分离到69株乳酸菌,鉴定为7个属11个种及亚种,其中Streptococcus salivarius为优势菌,通过初步筛选获得2株具有潜在益生特性的双歧杆菌：Bifidobacterium longum IMAU99727和Bifidobacterium animalis subsp.lactis IMAU99728,有望作为新的母乳源益生菌进行其他益生特性研究和开发利用。     

Isolation of bacteriocinogenic Lactobacillus plantarum strains from ben saalga, a traditional fermented gruel from Burkina Faso

A collection of lactic acid bacteria isolated from ben saalga, a traditional fermented gruel from Burkina Faso, was screened for bacteriocin production. Seven isolates were selected for their broad antimicrobial spectra, which overall included strains of Bacillus cereus, Bacillus licheniformis, Enterococcus faecalis, Listeria innocua, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli and Salmonella enterica. Cluster analysis of RAPD-PCR patterns revealed that six of the isolates represent different strains. The six selected strains were identified as Lactobacillus plantarum by 16S rDNA sequencing, species-specific PCR and multiplex PCR of the recA gene. PCR amplification revealed the presence of genes of the plantaricin cluster described in L. plantarum C11. Among them, strain 5.2.2 carried the largest number of genes from this cluster.     

The locus responsible for production of plantaricin S,a class IIb bacteriocin produced by Lactobacillus plantarum LPCO10, is widely distributed among wild-type Lact. plantarum strains isolated from olive fermentations

The genes plsA and plsB encoding for production of plantaricin S (Pls), a two-peptide bacteriocin produced by Lactobacillus plantarum LPCO10, are commonly distributed among wild-type Lact. plantarum strains isolated from olive fermentations. Among 68 independent isolates from different olive processing plants in South Spain, 15 of them were shown to produce bacteriocins that were active against other lactic acid bacteria, as well as spoilage and pathogenic bacteria. On the basis of PCR amplification and hybridization with specific probes, the Pls operon was detected in all the bacteriocin producer strains but not in the non-producer ones. Purification and subsequent amino acid sequencing of the bacteriocin produced by some of the 15 isolates yielded both the alpha and beta peptides from Pls. These results suggest that bacteriocin production contributes an ecological advantage for the wild-type Lact. plantanum strains in the colonization of the spontaneous, traditional olive fermentation process.     

新疆传统乳品中产胞外多糖乳酸菌的筛选及益生特性的研究

以新疆塔城地区酸奶、酸马奶、鲜马奶样品为研究对象,采用传统的分离方法对样品中的乳酸菌进行分离,通过16S rRNA分析样品中微生物多样性,在优势菌株中通过苯酚-硫酸法筛选出高产胞外多糖的乳酸菌,测定菌株潜在的益生特性。结果显示,3份酸奶、4份酸马奶和3份鲜马奶共10份样品中分离鉴定出147株乳酸菌,优势菌株为屎肠球菌（Enterococcus faecium）、发酵乳杆菌（Lactobacillus fermentum）和植物乳杆菌（Lactobacillus plantarum）。筛选出3株高产胞外多糖的植物乳杆菌（菌株1-3,1-6,4-1）。潜在益生特性试验结果显示,3株菌均表现出一定的益生特性,其中菌株1-3较其他菌株有较强的肠道定植、降胆固醇能力和胆盐耐受性,其可作为潜在益生菌应用于功能性产品的开发。     

Bacteriocin-producing Lactobacillus strains isolated from poto poto, a Congolese fermented maize product, and genetic fingerprinting of their plantaricin operons

Thirty one bacteriocin-producing Lactobacillus isolates were identified among 135 lactobacilli isolated from the Congolese fermented maize product poto poto, during the preparation and from the finished product. Using species-specific PCR and 16S rRNA gene sequencing, 28 and 3 isolates were identified as L. plantarum and L. fermentum, respectively. Cluster analysis of RAPD-PCR fingerprints revealed two main groups (G1 and G2) plus the L. fermentum isolate C4-13. Group G1 contained 23 isolates with a similarity coefficient >74.5%, and could be divided in two subgroups (G1-1, G1-2) each with several branches, plus the L. plantarum isolate C11. Group G2 contained 8 isolates with a similarity coefficient >86%, with two main branches. Using PCR amplification with specific primers, several genes of the plantaricin cluster found in L. plantarum C11 were identified in the isolates. The number of genes that were detected varied between the strains. The L. fermentum isolate EC11 also contained the plnDEFG genes. PCR amplification of DNA from isolates with primers directed to the upstream and downstream region of the plantaricin cluster generated an amplicon identical to that obtained with DNA from the control strain L. plantarum WCFS1. Amplification products from the positive strains were used for restriction analysis with HindIII, EcoRI and KpnI in separate reactions. Cluster analysis of restriction profiles revealed high similarities for EcoRI and HindII digest profiles, and an identical profile for all KpnI digests. The L. fermentum EC11 isolate clustered with L. plantarum strains in a group with a high correlation coefficient. The results suggest a low degree of diversity in the plantarincin gene cluster. However, other strains that tested positive for individual plantaricin genes may present great heterogeneity in the plantaricin operons. Because of their broad spectra of inhibition (including Escherichia coli, Salmonella enterica, Enterobacter aerogenes, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis), isolates from the present study could be used to improve the safety and storage stability of poto poto.     

四川牦牛奶和曲拉中九株乳酸菌的分子鉴定

为准确鉴定分离自我国四川鲜牦牛奶和曲拉中9株乳酸菌到种水平,作者运用16S rDNA序列分析、recA基因特异扩增和hsp60-RFLP等多种分子技术对分离自我国四川地区鲜牦牛奶和曲拉中的9株乳酸菌进行分类鉴定。结果证实,16S rDNA序列分析法可将9株乳酸菌初步归类为植物乳杆菌群(4株),肠膜明串珠菌(4株),瑞士乳杆菌(1株)。由于16S rDNA序列分析法不能区分植物乳杆菌和戊糖乳杆菌,为了进一步鉴定植物乳杆菌群中的4株菌,继续采用recA基因特异扩增和hsp60-RFLP技术对其细分,结果表明recA基因特异扩增和hsp60-RFLP的方法均能很好地把植物乳杆菌群中的4株菌鉴定到种水平,且均为植物乳杆菌。     

新疆喀什地区维吾尔族群体不同哺乳期母乳中乳杆菌的种群多样性

采用改良的MRS培养基进行菌株分离,多重聚合酶链式反应指纹图谱去重与16S rRNA同源性分析鉴定相结合,并通过MEGA V5.0建树对新疆喀什地区维吾尔族群体不同哺乳期母乳中乳杆菌的组成及变化进行研究。从45?个母乳样品中共分离出229?株乳酸菌,分别隶属于乳酸杆菌属（Lactobacillus）、肠球菌属（Enterococcus）和链球菌属（Streptococcus）,其中96.1%的菌株隶属Lactobacillus spp.,分别为短乳杆菌（L. brevis）75?株、发酵乳杆菌（L. fermentum）82 株、口乳杆菌（L. ori）37?株、阴道乳杆菌（L. vaginalis）4?株和加氏乳杆菌（L. gasseri）22?株。其中L. vaginalis是比较罕见的菌种。新疆喀什地区维吾尔族群体母乳中乳杆菌种群组成丰富,不同哺乳期母乳中乳杆菌变化趋势明显;成熟乳样品中分离的乳杆菌种群结构相对初乳和晚乳的要复杂,其中5?d～5?个月哺乳期的母乳中乳杆菌种群结构最为复杂,为母乳中益生菌资源的开发提供依据。     

Evaluation of Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 as probiotics by using a gastro-intestinal model with infant milk formulations as substrate

Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 produce bacteriocins with activity against a number of Gram-positive and Gram-negative bacteria. Both strains survived intestinal conditions simulated in a gastro-intestinal model (GIM) with infant milk formulations as substrate and prevented the growth of Listeria monocytogenes ScottA. The strains are inhibited by the antibiotics amoxicillin, cefadroxil, roxithromycin and doxycycline, anti-inflammatory medicaments containing meloxicam, ibuprofen and sodium diklofenak, and analgesics containing paracetamol, codeine phosphate and promethazine. Strain 423 is sensitive to vancomycin and does not contain genes encoding gelatinase, cell aggregation substance (AS), adhesion to collagen (Ace), enterococcus surface protein (Esp), Enterococcus faecalis endocarditis antigen (EfaAfs), cytolysin and non-cytolysin (beta-hemolysin III). Genes encoding AS, cytolysin and non-cytolysin (beta-hemolysin III) were amplified from the genome of strain ST4SA. Survival of strains ST4SA and 423 improved when used as combined cultures in the GIM and compared well with the survival of commercially available probiotics subjected to the same conditions.     

西藏地区发酵牛乳中产细菌素乳酸菌的筛选及鉴定

从西藏地区藏族传统发酵乳中分离乳酸菌,采用生理生化特性和16S基因序列同源性分析对其进行鉴定,通过双层琼脂平板扩散法筛选具有抑菌活性的菌株。结果表明,共分离37株乳酸菌,其中,乳杆菌属(Lactobacillus)35株、明串珠菌属(Leuconostoc)2株;35株乳酸杆菌为Lactobacillus casei 16株、Lactobacillus paracasei 7株、Lactobacillus plantarum 4株、Lactobacillus fermentum 2株、L.delbrueckii subsp.bulgaricus 2株、Lactobacillus helveti-cus 1株、Lactobacillus diolivorans 3株;7株L.casei和L.paracasei的发酵上清液对3株细菌指示菌表现出明显抑制作用,所有菌株对真菌无抑菌活性;在排除有机酸、H2O2等的干扰和经蛋白酶K处理后,初步确定7株乳酸菌发酵上清液中的抑菌物质为细菌素。     

传统乳制品中潜在益生植物乳杆菌的体外筛选

对从内蒙古牧区乳制品中分离到的29株植物乳杆菌的耐酸性（pH=3．0）、耐胆汁性（质量浓度3g／L牛胆酸钠）、抗菌活性进行了系统性研究。结果表明，有10株菌具有较好的耐酸耐胆盐特性，结合抗菌特性及黏附性试验结果，菌株WZ47-1，WZ32-2-1和WH13-2有可能成为潜在益生菌菌株。