Evaluation of lactic acid bacteria autolysate for the supplementation of lactic acid bacteria fermentation

At the end of culture in a carbon-limited medium, i.e. the best conditions for subsequent autolysis, lactic acid bacteria were harvested and autolysed at 50 °C for 24 h. The resulting supernatant was then successfully tested as a substitute for industrial yeast extract for the supplementation of whey permeate and its conversion into lactic acid: for almost equivalent total nitrogen amounts of both supplements, the same growth and production rates were recorded.     

Screening for antiproliferative effects of cellular components from lactic acid bacteria against human cancer cell lines

Whole cells, cytoplasms and peptidoglycans of ten different lactic acid bacteria (LAB) were tested for in vitro cytotoxicity on diverse cancer cell lines using the ³H-thymidine incorporation assay. The peptidoglycans and cytoplasm fractions, as well as heat-killed whole cells of LAB, had significant antiproliferative activities against several cancer cell lines. In particular, the cytoplasm fractions exhibited marked direct antiproliferative activities against colon and gastric cancer cell lines, whereas the peptidoglycans retarded growth of colon and bladder cancer cell lines. The cytoplasm fractions of Bifidobacterium longum and Lactococcus lactis ssp. lactis inhibited proliferation of two cancer cell lines by 50% at 33 and 23 g ml^–1 for SNUC2A (a human colon adenocarcinoma cell line) and 17 and 11 g ml^–1 for SNU-1 (a human gastric cancer cell line), respectively.     

Characterization of sourdough lactic acid bacteria based on genotypic and cell-wall protein analyses

AIMS: To evaluate the effectiveness of two independent methods in differentiating a large population of lactic acid bacteria (LAB) isolated from wheat flours and sourdoughs and to correlate eventual differences/similarities among strains with their geographical origin and/or process parameters. METHODS AND RESULTS: One hundred fifty strains belonging to Lactobacillus spp. and Weissella spp., plus eight type strains, one for each species, and two unidentified isolates, were characterized by randomly amplified polymorphic DNA (RAPD) and SDS-PAGE of cell-wall proteins. The RAPD analysis separated the eight type strains but did not always assign all the strains of a species to the same group, while SDS-PAGE cell-wall protein profiles were species-specific. Frequently, strains isolated from sourdoughs of the same geographical origin or produced by similar raw material/process parameters showed similar RAPD and/or cell-wall profiles. CONCLUSIONS: The combined use of the RAPD and cell-wall protein analysis represents a useful tool to classify large adventitious microbial populations and to discriminate the diversity of the strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents a typing of a large collection of flour/sourdough LAB and provides evidence of the advantage of using two independent methods in the classification and traceability of microorganisms.     

Key role of teichoic acids on aflatoxin B1 binding by probiotic bacteria

Aims:  To assess the ability of five probiotic bacteria to bind aflatoxin B₁ and to determine the key role of teichoic acids in the binding mechanism.
Methods and Results:  The strains were incubated in aqueous solutions containing aflatoxin B₁ (AFB₁). The amount of free toxin was quantified by HPLC. Stability of the bacteria–aflatoxin complex was evaluated by repeated washes with buffer. In order to understand the binding process, protoplasts, spheroplasts and cell wall components of two strains were analysed to assess their capacity to bind AFB₁. Additionally, the role of teichoic acids in the AFB₁ binding process was assessed. Lactobacillus reuteri strain NRRL14171 and Lactobacillus casei strain Shirota were the most efficient strains for binding AFB₁. The stability of the AFB₁–bacteria complex appears to be related to the binding ability of a particular strain; AFB₁ binding was also pH-dependent. Our results suggest that teichoic acids could be responsible for this ability.
Conclusions:  Our results provide information concerning AFB₁ binding by previously untested strains, leading to enhanced understanding of the mechanism by which probiotic bacteria bind AFB₁.
Significance and Impact of the Study:  Our results support the suggestion that some probiotic bacteria could prevent absorption of aflatoxin from the gastrointestinal tract.     

Genomics of lactic acid bacteria

Lactic acid bacteria (LAB) are found to occupy a variety of ecological niches including fermented foods as well as mucosal surfaces of humans and other vertebrates. This review is based on the genomic content of LAB that is responsible for the functional and ecological diversity of these bacteria. These genomes reveal an ongoing process of reductive evolution as the LAB have specialized to different nutritionally rich environments. Species-to-species variation in the number of pseudogenes as well as genes directing nutrient uptake and metabolism reflects the adaptation of LAB to food matrices and the gastrointestinal tract. Although a general trend of genome reduction was observed, certain niche-specific genes appear to be recently acquired and appear on plasmids or adjacent to prophages. Recent work has improved our understanding of the genomic content responsible for various phenotypes that continue to be discovered, as well as those that have been exploited by man for thousands of years.     

Multi-domain, cell-envelope proteinases of lactic acid bacteria

The multi-domain, cell-envelope proteinases encoded by the genes prtB of Lactobacillus delbrueckii subsp. bulgaricus, prtH of Lactobacillus helveticus, prtP of Lactococcus lactis, scpA of Streptococcus pyogenes and csp of Streptococcus agalactiae have been compared using multiple sequence alignment, secondary structure prediction and database homology searching methods. This comparative analysis has led to the prediction of a number of different domains in these cell-envelope proteinases, and their homology, characteristics and putative function are described. These domains include, starting from the N-terminus, a pre-pro-domain for secretion and activation, a serine protease domain (with a smaller inserted domain), two large middle domains A and B of unknown but possibly regulatory function, a helical spacer domain, a hydrophilic cell-wall spacer or attachment domain, and a cell-wall anchor domain. Not all domains are present in each cell-envelope proteinase, suggesting that these multi-domain proteins are the result of gene shuffling and domain swapping during evolution.     

Peptidases and amino acid catabolism in lactic acid bacteria

The conversion of peptides to free amino acids and their subsequent utilization is a central metabolic activity in prokaryotes. At least 16 peptidases from lactic acid bacteria (LAB) have been characterized biochemically and/or genetically. Among LAB, the peptidase systems of Lactobacillus helveticus and Lactococcus lactis have been examined in greatest detail. While there are homologous enzymes common to both systems, significant differences exist in the peptidase complement of these organisms. The characterization of single and multiple peptidase mutants indicate that these strains generally exhibit reduced specific growth rates in milk compared to the parental strains. LAB can also catabolize amino acids produced by peptide hydrolysis. While the catabolism of amino acids such as Arg, Thr, and His is well understood, few other amino acid catabolic pathways from lactic acid bacteria have been characterized in significant detail. Increasing research attention is being directed toward elucidating these pathways as well as characterizing their physiological and industrial significance.     

乳酸菌的系统分类概况

乳酸菌是重要的益生菌资源,在食品、农业、化工业、医学等领域具有广阔的应用前景。目前,人们熟知的乳酸菌主要集中在乳杆菌属、双歧杆菌属、链球菌属、肠球菌属、乳球菌属、片球菌属和明串珠菌属等少数属种。为了拓宽人们对乳酸菌的认知,本文就乳酸菌的系统分类学进行阐述。在系统分类学上,乳酸菌分别隶属于厚壁菌门4纲7目18科39属653种和放线菌门2纲2目3科12属88种。最后,对乳酸菌的益生作用、安全性与有效来源、益生潜能的体外评价指标等进行简要的讨论。     

Exocellular polysaccharides produced by lactic acid bacteria

Abstract The production of homopolysaccharides (dextrans, mutans) and heteropolysaccharides by lactic acid bacteria, their chemical composition, their structure and their synthesis are outlined. Mutans streptococci, which include Streptococcus mutans and S. sobrinus produce soluble and insoluble α-glucans. The latter may contain as much as 90%α-1–3 linkages and possess a marked ability to promote adherence to the smooth tooth surface causing dental plaque. Dextrans produced by Leuconostoc mesenteroides are high molecular weight α-glucans having 1–6, 1–4 and 1–3 linkages, varying from slightly to highly branched; 1–6 linkages are predominant. Emphasis is put on exopolysaccharide producing thermophilic and mesophilic lactic acid bacteria, which are important in the dairy industry. The produced polymers play a key role in the rheological behaviour and the texture of fermented milks. One of the main problems in this field is the transitory nature of the thickening trait. This instability is not yet completely understood. Controversial results exist on the sugar composition of the slime produced, but galactose and glucose have always been identified with galactose predominating in most cases.     

一株能降解胆固醇的乳酸菌的选育

利用选择性培养基从成年人的粪便中分离出1株能降解胆固醇的乳酸菌,该菌能以胆固醇作为生长的唯一能源。在10％牛奶的发酵试验中,该菌能使牛奶发酵并凝固。在液体发酵中,胆固醇的降解率为34．6％。经初步鉴定,该菌为双歧杆菌（Bifidoacterium sp.)。     

代谢组学在乳酸菌研究中的应用

代谢组学是系统生物学的重要分支,因其高效、高通量等特点而广泛应用于食品科学、药物学等研究领域。本文概述了代谢组学的分离和检测技术,综述了代谢组学在乳酸菌鉴定、发酵调控、肠道菌群研究等方面中的应用,对代谢组学在乳酸菌研究中潜在的问题和未来发展趋势进行了讨论,期望为代谢组学在食品工业微生物中的应用提供参考。     

Boza, a natural source of probiotic lactic acid bacteria

Aims: To evaluate the probiotic properties of strains isolated from boza, a traditional beverage produced from cereals. Methods and Results: The strains survived low pH conditions (pH 3·0), grew well at pH 9·0 and were not inhibited by the presence of 0·3% (w/v) oxbile. Cytotoxicity levels of the bacteriocins, expressed as CC₅₀, ranged from 38 to 3776 μg ml^?1. Bacteriocin bacST284BZ revealed high activity (EC₅₀ = 735 μg ml^?1) against herpes simplex virus type 1. Growth of Mycobacterium tuberculosis was 69% repressed after 5 days in the presence of bacST194BZ. Various levels of auto‐cell aggregation and co‐aggregation with Listeria innocua LMG 13568 were observed. Adhesion of the probiotic strains to HT‐29 cells ranged from 18 to 22%. Conclusions: Boza is a rich source of probiotic lactic acid bacteria. All strains survived conditions simulating the gastrointestinal tract and produced bacteriocins active against a number of pathogens. Adherence to HT‐29 and Caco‐2 cells was within the range reported for Lactobacillus rhamnosus GG, a well‐known probiotic. In addition, the high hydrophobicity readings recorded define the strains as good probiotics. Significance and Impact of the Study: Boza contains a number of different probiotic lactic acid bacteria and could be marketed as a functional food product.     

一株产共轭亚油酸乳酸菌的鉴定及其特性

从酸菜汁中分离筛选到一株产共轭亚油酸(CLA)能力较高的乳酸菌。经鉴定,确定为植物乳杆菌Lactobacliius plantarum。微氧条件可提高CLA的产量,催化亚油酸(LA)生成CLA的酶受着LA的诱导。37℃对细胞生长和CLA生成最为有利。对数生长期为6～12h,18h后进入稳定期。在14～22h,CLA生成量快速增加,24h时达到最高值。该菌的培养物经萃取、甲酯化后,进行了气相色谱分离,生成的CLA产物为c9/t9,c11-CLA和t10,c12-CLA异构体的混合物。     

The production-influencing factors of extracellular polysaccharide (EPS) from a strain of lactic acid bacteria and EPS extraction

The influencing factors of extracellular polysaccharide (EPS) produced from a strain of lactic acid bacteria (LAB L15) were studied by using the phenol-H₂SO₄ method. It was demonstrated that the strain produced EPS at the most amount when it was incubated for 40–48 h and when the pH value was 4 under 30°C. Glucose was the most suitable carbon source for LAB-producing EPS. The rough EPS was obtained from L15 culture after centrifugation, dialysis, deprotein, decoloration, and ethanol-precipitation. The sample was at least composed of two polysaccharides that were completely different in molecular weight and the amount. The purified EPS was passed through the SephadexG-200 column and it showed that it was a sample purified by thin layer chromatography. __________ Translated from Microbiology, 2005, 32(4): 85–90 [译自: 微生物学通报, 2005, 32(4): 85–90]     

The production-influencing factors of extracellular polysacchadde(EPS) from a Strain of lactic acid bacteria and EPS extraction

The influencing factors of extracellular polysaccharide(EPS)produced from a strain of lactic acid bacteria(LAB L15)were studied by using the phenol-H2SO4 method.It was demonstrated that the strain produced EPS at the most amount when it was incubated for 40-48 h and when the pH value was 4 under 30℃.Glucose was the most suitable carbon source for LAB-producing EPS.The rough EPS was obtained from L15 culture after centrifugation,dialysis,deprotein,decoloration,and ethanol-precipitation.The sample was at least composed of two polysaccharides mat were completely different in molecular weight and the amount.The purified EPS was passed through the SephadexG-200 colunm and it showed that it was a sample purified by thin layer chromatography.     

乳酸菌对真菌毒素的脱毒作用研究进展

真菌毒素广泛存在于农业产品中,对人和动物的健康构成巨大威胁。乳酸菌作为一种公认安全的微生物,在食品生物减毒方面具有巨大的应用潜力,成本低廉且不会对食品品质及生态环境造成不良影响。文章主要根据近年来国内外研究进展,阐述乳酸菌对食品和饲料中几种常见真菌毒素的脱毒作用(抑制真菌生长、毒素的吸附和降解),关注乳酸菌在生物脱毒方面的实际应用,为乳酸菌在食品保鲜领域的应用提供理论指导。     

Anti-mutagenic and immuno-stimulatory properties of lactic acid bacteria

Statistically significant antigenotoxic activity was exerted by six of nine strains of lactic acid bacteria tested (Lactobacillus delbrueckii subsp. bulgaricus, Staphylococcus carnosus, Streptococcus thermophilus, L. rhamnosus, Enterococcus faecium and En. faecalis) against nitrovin and 2-aminofluorene in Salmonella typhimurium TA100 and TA97. The mutagenic activity of both mutagens was substantially decreased by viable bacteria; cells heated to 100°C for 15 min were ineffective. In vitro, En. faecium stimulated the basic metabolic activities of human neutrophils which were essential for their antimicrobial and cytotoxic activity, whereas stimulation of guinea-pig macrophages was not so effective. Similar immuno-stimulatory effects were observed with both viable and heat-inactivated bacteria.L. Ebringer, M. Lahitová and D. Michálková are with the Institute of Molecular and Subcellular Biology, Comenius University, Odborárske nám. 5, SK-81107 Bratislava, Slovakia. M. Fereník and L. Kaáni are with the Institute of Immunology, Faculty of Medicine, Comenius University, SK-81108 Bratislava, Slovakia.     

Resident lactic acid bacteria in raw milk Canestrato Pugliese cheese

AIMS: Investigation of the autochthonous lactic acid bacteria (LAB) population of the raw milk protected designation of origin Canestrato Pugliese cheese using phenotypic and genotypic methodologies. METHODS AND RESULTS: Thirty phenotypic assays and three molecular techniques (restriction fragment length polymorphism, partial sequencing of the 16S rRNA gene and recA multiplex PCR assay) were applied to the identification of 304 isolates from raw milk Canestrato Pugliese cheese. As a result, 168 of 207 isolates identified were ascribed to genus Enterococcus, 25 to Lactobacillus, 13 to Lactococcus and one to Leuconostoc. More in details among the lactobacilli, the species Lactobacillus brevis and Lactobacillus plantarum were predominant, including 13 and 10 isolates respectively, whereas among the lactococci, Lactococcus lactis subsp.cremoris [corrected] was the species more frequently detected (seven isolates). CONCLUSIONS: Except for the enterococci, phenotypic tests were not reliable enough for the identification of the isolates, if not combined to the genotype-based molecular techniques. The polyphasic approach utilized allowed 10 different LAB species to be detected; thus suggesting the appreciable LAB diversity of the autochthonous microbial population of the Canestrato Pugliese cheese. SIGNIFICANCE AND IMPACT OF THE STUDY: A comprehensive study of the resident raw milk Canestrato Pugliese cheese microbial population has been undertaken.     

Hydrolysis of soybean isoflavone glucosides by lactic acid bacteria

Lactobacillus delbrueckii subsp. delbrueckii KCTC 1047, grown in de Man, Rogosa and Sharpe (MRS) or soymilk media, completely hydrolyzed the isoflavone glucosides, genistin and daidzin at 50 g ml^–1, into their respective aglycones, genistein and daidzein within 30 min. Other lactic acid bacteria did not produce -glucosidase, the enzyme responsible for the hydrolysis of isoflavone glucosides, when cultured in MRS medium. Glucoside-hydrolyzing activity was induced in some lactic acid bacteria when cultured in soymilk medium. These strains hydrolyzed 70–80% of genistin into genistein and 25–40% of daidzin into daidzein.