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At least 649 records · Page 36Linked to original sources
PubMed1993

Cloning, DNA sequence analysis and partial characterization of pepN, a lysyl aminopeptidase from Lactobacillus delbrückii ssp. lactis DSM7290.

In cell extracts of Lactobacillus delbrückii ssp. lactis DSM7290 a peptidase with the ability to hydrolyse Phe-beta-naphthylamide (Phe-beta-NA) and His-beta-NA could be detected. Escherichia coli lacking the enzyme activity in an enzymic plate assay was used to screen high-copy-number and low-copy-number plasmid libraries of size-fractionated Lactobacillus DNA. Clones with the desired phenotype were detected, and the gene, designated pepN, was further subcloned and sequenced. A large open reading frame of 2529 nucleotides is predicted to encode a protein of 843 amino acids (95358 Da). Comparison of the pepN gene from Lb. delbrückii ssp. lactis DSM7290 indicates that it is homologous to genes of the family of Zn(2+)-metallohydrolases and PepN shows identity with the active centre Zn(2+)-binding motif of these enzymes. The substrate Lys-beta-NA is more effectively cleaved than Phe-beta-NA or His-beta-NA which were used for screening in E. coli. The cloned pepN gene was efficiently overexpressed in E. coli and subcloning of the gene in Lactobacillus casei resulted in a moderate overexpression of approximately 20-fold. The pepN gene product was purified from the pepN-deficient E. coli strain CM89, using the substrate Lys-p-nitroanilide (Lys-NH-Ph) in the assay procedure. In a four-step procedure including streptomycin sulfate precipitation, anion-exchange chromatography and gel filtration the peptidase was purified to electrophoretic homogeneity.

Amino Acid Sequence↗
PubMed1983

Potentiation of large intestinal tumorigenicity of cycasin derivative by high-fat diet and Lactobacillus in germfree mice.

Large intestinal tumorigenesis was investigated in germ free (GF) mice following the administration of methylazoxymethanol (MAM) acetate, lard, and Lactobacillus arabinosus. Twelve weekly doses of 0.3 mg/10 g MAM acetate given to groups of male weanling, germfree ICR mice, fed either low-fat (LF) chow alone, or chow with added lard (HF). Additional groups of mice, on the low-fat diet, were monocontaminated with Lactobacillus arabinosus (GA) and they and their GF controls were given 12 weekly doses of 0.2 mg/10 g of MAM acetate. The HF and LF groups were sacrificed after 123 days, and the GA and GF groups after 214 days. Serial sections of the entire large intestine of the MAM acetate-treated mice revealed many more sessile polyps in the mice maintained on the high-fat diet. In the mice monocontaminated with Lactobacillus, numerous tumors were found, of which 25 percent were invasive. In addition, the polyps found in these animals were generally considerably larger than the polyps found in the GF mice. Atypia occurred in the large intestinal mucosa in all groups. Lard enhanced the large intestinal polypogenesis induced by MAM acetate in germfree mice. Gnotobiosis potentiated the formation of sessile polyps, and carcinogenesis.

Animals↗
PubMed1996

Antagonistic effects of intestinal Lactobacillus isolates on pathogens of chicken.

Twelve Lactobacillus strains isolated from chicken intestine, which demonstrated a strong and moderate capacity to adhere to the ileal epithelial cells in vitro, were used to investigate their inhibitory ability against five strains of salmonella, i.e. Salmonella enteritidis 935/79, Salm. pullorum, Salm. typhimurium, Salm. blockley and Salm. enteritidis 94/448, and three serotypes of Escherichia coli, viz. E. coli O1:K1, O2:K1 and O78:K80. The results showed that all the 12 Lactobacillus isolates were able to inhibit the growth of the five strains of salmonella, and the three strains of E. coli in varying degrees. Generally, they were more effective in inhibiting the growth of salmonella than E. coli. Inhibition of the pathogenic bacteria was probably due to the production of organic acids by the Lactobacillus isolates.

Animals↗
PubMed1994

Cloning and expression of the plasmid encoded beta-D-galactosidase gene from a Lactobacillus plantarum strain of dairy origin.

The beta-galactosidase (beta-Gal) gene from Lactobacillus plantarum C3.8 was cloned and expressed in Lactococcus lactis and Escherichia coli. Hybridization experiments indicated that the gene is located on a plasmid and is present in other strains of Lactobacillus plantarum. Its sequence is very similar to a Leuconostoc lactis beta-Gal gene. Expression of the gene, both in Lactobacillus plantarum and in Lactococcus lactis, was four-fold higher in cells growth in lactose compared to those grown in glucose. The presence of the beta-Gal gene in Lactococcus lactis allowed this bacterium to be efficient in clotting milk.

Amino Acid Sequence↗
PubMed2000

Purification and characterization of a surface-binding protein from Lactobacillus fermentum RC-14 that inhibits adhesion of Enterococcus faecalis 1131.

Lactobacilli have been shown to be important in the maintenance of the healthy urogenital flora. One strain, Lactobacillus fermentum RC-14, releases surface-active components which can inhibit adhesion of uropathogenic bacteria. Using a quantitative method for determining inhibition of adhesion, a protein with high anti-adhesive properties against Enterococcus faecalis 1131 was purified. The N-terminal sequence of the 29-kDa protein was identical to that of a collagen-binding protein from Lactobacillus reuteri NCIB 11951, and exhibited close homology with a basic surface protein from L. fermentum BR11. The results suggest that this anti-adhesive cell surface protein of Lactobacillus could protect against uropathogens by preventing their adhesion. the Federation of European Microbiological Societies.

Amino Acid Sequence↗
PubMed1987

Can the presence of erupting or exfoliating teeth increase the salivary lactobacillus count?

The salivary lactobacillus count was assessed during school attendance among 1734 7-15-yr-old children and related to the number of teeth erupting within each age group. The prevalence of extremely low lactobacillus counts (less than or equal to 10(3)) was inversely correlated with the number of erupting teeth (P less than 0.001) but the differences were small. High counts differed only by 7% between ages with the lowest and the highest mean number of erupting or exfoliating teeth tend to elevate the lactobacillus count in individuals. On a population level the effect is small.

Adolescent↗
PubMed1995

Effect of Lactobacillus rhamnosus strain GG (ATCC 53103) on the growth of Streptococcus sobrinus in vitro.

Lactobacillus GG, a recently characterized L. rhamnosus GG strain (ATCC 53103), has been shown to exert inhibitory activity against a variety of bacterial species, including streptococci. We isolated and studied the effect of the inhibitory substance of Lactobacillus GG on some oral streptococci. The inhibitory activity of the isolated substance was weak, but some growth inhibition was observed in Streptococcus sobrinus pretreated with the substance in comparison with untreated controls. Zones of growth inhibition on agar plates were apparent only at pH values below 5, indicating that the inhibitory activity was restricted to a low pH range. Growth curve experiments showed a statistically significant inhibition between series with and without the isolated substance (P < 0.05). The ultrastructure of S. sobrinus was not affected when treated with the inhibitory substance. The Lactobacillus GG itself did not ferment sucrose. The results offer interesting perspectives for future research focusing on the protective function of normal flora and in the attempt to replace harmful bacterial species in oral microflora with less harmful ones.

Antibiosis↗
PubMed2006

Engineered vaginal lactobacillus strain for mucosal delivery of the human immunodeficiency virus inhibitor cyanovirin-N.

Women are at significant risk of human immunodeficiency virus (HIV) infection, with the cervicovaginal mucosa serving as a major portal for virus entry. Female-initiated preventatives, including topical microbicides, are urgently needed to help curtail the HIV/AIDS pandemic. Here we report on the development of a novel, live microbicide that employs a natural vaginal strain of Lactobacillus jensenii engineered to deliver the potent HIV inhibitor cyanovirin-N (CV-N). To facilitate efficient expression of CV-N by this bacterium, the L. jensenii 1153 genome was sequenced, allowing identification of native regulatory elements and sites for the chromosomal integration of heterologous genes. A CV-N expression cassette was optimized and shown to produce high levels of structurally intact CV-N when expressed in L. jensenii. Lactobacillus-derived CV-N was capable of inhibiting CCR5-tropic HIV(BaL) infectivity in vitro with a 50% inhibitory concentration of 0.3 nM. The CV-N expression cassette was stably integrated as a single copy into the bacterial chromosome and resolved from extraneous plasmid DNA without adversely affecting the bacterial phenotype. This bacterial strain was capable of colonizing the vagina and producing full-length CV-N when administered intravaginally to mice during estrus phase. The CV-N-producing Lactobacillus was genetically stable when propagated in vitro and in vivo. This work represents a major step towards the development of an inexpensive yet durable protein-based microbicide to block the heterosexual transmission of HIV in women.

Administration, Intravaginal↗
PubMed1999

In vitro activities of ketolide HMR3647, macrolides, and other antibiotics against Lactobacillus, Leuconostoc, and Pediococcus isolates.

Testing of susceptibility to 13 antibiotics was performed with 90 isolates of Lactobacillus, Leuconostoc, and Pediococcus. MICs at which 90% of the isolates tested were inhibited by HMR3647, erythromycin, and ciprofloxacin were 0.015, 0.125 and 32 microg/ml, respectively. The penicillin MIC was > or = 16 microg/ml against 26.2% of the studied Lactobacillus sp. isolates and 50% of Lactobacillus plantarum. HMR3647 showed excellent activity against these genera.

Anti-Bacterial Agents↗
PubMed1983

Viricidal effects of Lactobacillus and yeast fermentation.

The survival of selected viruses in Lactobacillus- and yeast-fermented edible waste material was studied to determine the feasibility of using this material as a livestock feed ingredient. Five viruses, including Newcastle disease virus, infectious canine hepatitis virus, a porcine picornavirus, frog virus 3, and bovine virus diarrhea, were inoculated into a mixture of ground food waste (collected from a school lunch program) containing Lactobacillus acidophilus. Mixtures were incubated at 20, 30, and 40 degrees C for 216 h. In a second trial, four viruses, including Newcastle disease virus, infectious canine hepatitis virus, frog virus 3, and a porcine picornavirus, were inoculated into similar edible waste material containing Saccharomyces cerevisiae. Mixtures were incubated at 20 and 30 degrees C for 216 h. Samples were obtained daily for quantitative (trial 1) and qualitative (trial 2) virus isolation. Temperature, pH, and redox potential were monitored. Controlled pH and temperature studies were also done and compared with the inactivation rates in the fermentation processes. In trial 1 (Lactobacillus fermentation), infectious canine hepatitis virus survived the entire test period in the fermentation process but was inactivated below pH 4.5 in the controlled studies. Newcastle disease virus was inactivated by day 8 in the fermentation process and appeared to be primarily heat sensitive and secondarily pH sensitive in the controlled studies. The porcine picornavirus survived the fermentation process for 8 days at 20 degrees C but was inactivated more rapidly at 30 and 40 degrees C. The controlled studies verified these findings. Frog virus 3 was inactivated by day 3 in the fermentation process and appeared to be sensitive to low pH in the controlled studies. Bovine virus diarrhea was rapidly inactivated in the fermentation process (less than 2 h) and was pH and temperature sensitive. In trial 2 (yeast fermentation), infectious hepatitis virus survived the entire test period in the fermentation process. Newcastle disease virus was inactivated by day 7 at 20 degrees C and day 6 at 30 degrees C. The porcine picornavirus was inactivated by day 7 at 30 degrees C but survived the entire test period at 20 degrees C. Frog virus 3 was inactivated by day 3 at 20 degrees C and day 2 at 30 degrees C.

Fermentation↗
PubMed1988

Genetic transformation in Lactobacillus sp. strain 100-33 of the capacity to colonize the nonsecreting gastric epithelium in mice.

Lactobacillus isolates able to colonize the surfaces of the nonsecreting epithelia in the stomachs of monoassociated ex-germfree mice were derived from Lactobacillus acidophilus 100-33. Strain 100-33 was originally isolated from pig feces and is unable to colonize the murine gastric epithelium. In experiments involving attempts genetically to transform the capacity to colonize the epithelium, cells of strain 100-33 were treated with muralytic enzymes and mixed with polyethylene glycol and genomic or plasmid DNA extracted from Lactobacillus fermentum RI. Strain RI was originally isolated from a conventional mouse and has the capacity to colonize the nonsecreting gastric epithelium. The mixtures containing cells, polyethylene glycol, and DNA were plated on a regeneration medium. After overnight incubation, the cells were washed from the plates and introduced by gastric gavage into germfree mice. Only mice that received regenerated 100-33 cells previously mixed with genomic DNA from strain RI had layers of gram-positive bacteria on the keratinized epithelia of their stomachs. Six isolates cultured from the washed gastric tissues of these animals were characterized. When a culture of each or a pool of cultures of the six were orally administered to germfree mice, layers of gram-positive bacterial cells were visible on the keratinized gastric epithelia of the animals within 1 to 3 weeks. Cells of all six, but not of strain 100-33, reacted with antibody made in rabbits to L. fermentum RI cells, as determined by an enzyme-linked immunosorbent assay. Nevertheless, all six had fermentation profiles identical to that of strain 100-33.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Characterization of a Lactobacillus strain producing white crystals on cheddar cheese.

From an enrichment culture of white-crystal deposits from aged Cheddar cheese, an atypical Lactobacillus strain was characterized. The new isolate is facultatively heterofermentative, has a G + C content of 40 mol%, and produces D and L isomers of lactic acid. The strain had a limited ability to ferment carbohydrates. It utilized fructose, galactose, glucose, lactose, maltose, mannose, and ribose but was negative for esculin, gluconate, citrate, and several other carbon sources. The isolate also had low DNA-DNA homologies with strains of Lactobacillus casei and Lactobacillus plantarum. Cheese prepared with milk containing the isolate developed white crystals during curing. Formation of copious D-lactate from unknown substrates during curing probably caused the white-crystal deposits. The strain has been deposited in the American Type Culture Collection (ATCC 49178).

Carbohydrate Metabolism↗
PubMed1990

Lactobacillus succession in the piglet digestive tract demonstrated by plasmid profiling.

Plasmid profiling was used to distinguish strains of lactobacilli inhabiting the digestive tract of piglets and the feces of sows. Fifteen plasmid profile types were detected among 328 isolates of lactobacilli. Plasmid profiling of lactobacilli permitted the following conclusions to be made: the maternal feces were a major source of lactobacilli colonizing the piglet digestive tract; the lactobacillus population of the gastric region of the piglet digestive tract was composed of lactobacillus strains different from those present in the rectal population; and a lactobacillus succession was observed in the digestive tract of piglets drawn from a single litter, and one plasmid profile type became dominant in the gastric region of these animals.

Animals↗
PubMed1992

Cloning and expression of a conjugated bile acid hydrolase gene from Lactobacillus plantarum by using a direct plate assay.

The conjugated bile acid hydrolase gene from the silage isolate Lactobacillus plantarum 80 was cloned and expressed in Escherichia coli MC1061. For the screening of this hydrolase gene within the gene bank, a direct plate assay developed by Dashkevicz and Feighner (M. P. Dashkevicz and S. D. Feighner, Appl. Environ. Microbiol. 53:331-336, 1989) was adapted to the growth requirements of E. coli. Because of hydrolysis and medium acidification, hydrolase-active colonies were surrounded with big halos of precipitated, free bile acids. This phenomenon was also obtained when the gene was cloned into a multicopy shuttle vector and subsequently reintroduced into the parental Lactobacillus strain. The cbh gene and surrounding regions were characterized by nucleotide sequence analysis. The deduced amino acid sequence was shown to have 52% similarity with a penicillin V amidase from Bacillus sphaericus. Preliminary characterization of the gene product showed that it is a cholylglycine hydrolase (EC 3.5.1.24) with only slight activity against taurine conjugates. The optimum pH was between 4.7 and 5.5. Optimum temperature ranged from 30 to 45 degrees C. Southern blot analysis indicated that the cloned gene has similarity with genomic DNA of bile acid hydrolase-active Lactobacillus spp. of intestinal origin.

Amidohydrolases↗
PubMed1992

Cloning, sequence, and phenotypic expression of katA, which encodes the catalase of Lactobacillus sake LTH677.

Lactobacillus sake LTH677 is a strain, isolated from fermented sausage, which forms a heme-dependent catalase. This rare property is highly desirable in sausage fermentation, as it prevents rancidity and discoloration caused by hydrogen peroxide. A gene bank containing MboI fragments of chromosomal DNA from Lactobacillus sake LTH677 in Escherichia coli plasmid pBR328 was constructed. The catalase gene was cloned by heterologous complementation of the Kat- phenotype of E. coli UM2. The catalase structural gene, designated katA, was assigned to a 2.3-kb region by deletion analysis of the originally cloned fragment in plasmid pHK1000. The original chromosomal arrangement was determined by Southern hybridization. Protein analysis revealed that the catalase subunit has a molecular size of 65,000 Da and that the active catalase possesses a hexameric structure. The molecular size of the subunit deduced from the nucleotide sequence was determined to 54,504 Da. The N-terminal amino acid sequence of the 65,000-Da protein corresponded to the one deduced from the DNA sequence. After recloning of katA in the E. coli-Lactococcus shuttle vector pGKV210, the gene was successfully transferred and phenotypically expressed in Lactobacillus casei, which is naturally deficient in catalase activity.

Amino Acid Sequence↗
PubMed1992

Protein-mediated adhesion of Lactobacillus acidophilus BG2FO4 on human enterocyte and mucus-secreting cell lines in culture.

The adhesion of Lactobacillus acidophilus BG2FO4, a human stool isolate, to two human enterocytelike cell lines (Caco-2 and HT-29) and to the mucus secreted by a subpopulation of mucus-secreting HT29-MTX cells was investigated. Scanning electron microscopy revealed that the bacteria interacted with the well-defined apical microvilli of Caco-2 cells without cell damage and with the mucus secreted by the subpopulation of HT29-MTX cells. The adhesion to Caco-2 cells did not require calcium and involved an adhesion-promoting factor that was present in the spent supernatant of L. acidophilus cultures. This factor promoted adhesion of poorly adhering human Lactobacillus casei GG but did not promote adhesion of L. casei CNRZ 387, a strain of dairy origin. The adherence components on the bacterial cells and in the spent supernatant were partially characterized. Carbohydrates on the bacterial cell wall appeared to be partly responsible for the interaction between the bacteria and the extracellular adhesion-promoting factor. The adhesion-promoting factor was proteinaceous, since trypsin treatment dramatically decreased the adhesion of the L. acidophilus strain. The adhesion-promoting factor may be an important component of Lactobacillus species that colonize the gastrointestinal tract.

Bacterial Adhesion↗
PubMed1993

Derivation of DNA probes for enumeration of a specific strain of Lactobacillus acidophilus in piglet digestive tract samples.

Four DNA probes were derived that hybridized specifically to DNA from Lactobacillus acidophilus O. The probes were constructed by randomly cloning lactobacillus DNA in plasmid vector pBR322. Two of the probes (pSR1 and pSR2) were composed of vector and plasmid DNA inserts (3.6 and 1.6 kb, respectively); the others (pSR3 and pSR4) were composed of vector and chromosomally derived inserts (6.9 and 1.4 kb, respectively). The probes were used to enumerate, by colony hybridization, strain O in digestive tract samples collected from piglets inoculated 24 hours previously with a culture of the strain. The probes did not hybridize to DNA from lactobacilli inhabiting the digestive tract of uninoculated piglets. Strain O made up about 10% of the total lactobacillus population of the pars esophagea and about 20% of the population in other digestive tract samples.

Animals↗
PubMed1994

Kinetic studies of the action of lactacin F, a bacteriocin produced by Lactobacillus johnsonii that forms poration complexes in the cytoplasmic membrane.

The bacteriocin lactacin F is bactericidal against Lactobacillus delbrueckii, Lactobacillus helveticus, and Enterococcus faecalis. Activity against L. delbrueckii was recently shown to be dependent on two peptides, LafA and LafX, which are encoded within the lactacin F operon (T. R. Klaenhammer, FEMS Microbiol. Rev. 12:39-87, 1993). It has been proposed that two peptides form an active lactacin F complex. In this study, the action of lactacin F against E. faecalis ATCC 19443 and the effects of various environmental parameters were investigated in detail. Addition of lactacin F induced the loss of K+ from cells of L. delbrueckii, Lactobacillus johnsonii 88-4, and E. faecalis, while the lactacin F producer L. johnsonii VPI 11088 was not affected by the bacteriocin. Lactacin F caused an immediate loss of cellular K+, depolarization of the cytoplasmic membrane, and hydrolysis of internal ATP in E. faecalis. Lactacin F induced loss of K+ in 3,3',4',5-tetrachlorosalicylanilide-treated cells, indicating that pores are formed in the absence of a proton motive force. ATP hydrolysis was not due to dissipation of the proton motive force but was most likely caused by efflux of inorganic phosphate, resulting in a shift of the ATP hydrolysis equilibrium. Action of lactacin F was optimal at acidic pH values and was reduced in the presence of di- and trivalent cations. The lanthanide gadolinium (Gd3+) prevented action of lactacin F completely at a concentration of 0.2 mM. Lactacin F-induced loss of cell K+ was severely reduced at low temperatures, presumably as a result of increased ordering of the lipid hydrocarbon chains in the cytoplasmic membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗