Gasseri Strains Research Articles

Lactobacillus gasseri requires peptides, not proteins or free amino acids, for growth in milk

Lactobacillus gasseri is a widespread commensal lactic acid bacterium inhabiting human mucosal niches and has many beneficial effects as a probiotic. However, L. gasseri is difficult to grow in milk, which hurts usability for the food industry. It had been previously reported that supplementation with yeast extract or proteose peptone, including peptides, enables L. gasseri to grow well in milk. In this study, our objective was to confirm peptide requirement of L. gasseri and evaluate efficacy of peptide release by enzymatic proteolysis on growth of L. gassei in milk. Three strains of L. gasseri did not grow well in modified DeMan, Rogosa, Sharpe broth without any nitrogen sources (MRS-N), but addition of a casein-derived peptide mixture, tryptone, promoted growth. In contrast, little effect was observed after adding casein or a casein-derived amino acid mixture, casamino acids. These results indicate that L. gasseri requires peptides, not proteins or free amino acids, among milk-derived nitrogen sources for growth. Lactobacillus gasseri JCM 1131T hardly had growth capacity in 6kinds of milk-based media: bovine milk, human milk, skim milk, cheese whey, modified MRS-N (MRSL-N) supplemented with acid whey, and MRSL-N supplemented with casein. Moreover, treatment with digestive proteases, particularly pepsin, to release peptides made it grow well in each milk-based medium. The pepsin treatment was the most effective for growth of strain JCM 1131T in skim milk among the tested food-grade proteases such as trypsin, α-chymotrypsin, calf rennet, ficin, bromelain, and papain. As well as strain JCM 1131T, pepsinolysis of milk improved growth of other L. gasseri strains and some strains of enteric lactobacilli such as Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus johnsonii, and Lactobacillus reuteri. These results suggest that some relatives of L. gasseri also use peptides as desirable nitrogen sources, and that milk may be a good supplier of nutritious peptides to enteric lactobacilli including L. gasseri after peptic digestion in the gastrointestinal tract. This is the first report showing peptide requirement of L. gasseri and efficacy of pepsinolysis on the growth of L. gasseri and its relatives in milk. This study would contribute to increasing usability of L. gasseri and its relatives as probiotics in dairy foods.

Probiotic and cultural characteristic of strain Lactobacillus gasseri 4/13 of human origin

Lactobacillus gasseri within the Lactobacillus acidophilus group is a major species in the human microflora. The potential probiotic properties of a L. gasseri strain of human origin were evaluated. Out of 17 studied L. gasseri strains, L. gasseri 4/13 showed the highest immunomodulatory effect (induction of interferon gamma measured by sandwich enzyme-linked immunosorbent assay) in Balb/c mouse splenocytes in vitro and the highest rate of adhesion to Caco-2 human epithelial cells. The strain also reduced the concentration of cholesterol in the growth medium by 65% as compared with the initial concentration (measured spectrophotometrically). These probiotic properties indicate that L. gasseri 4/13 could prove an attractive concentrated adjunct monoculture in the production of new functional foods. To obtain a freeze-dried bacterial concentrate from L. gasseri 4/13, the influence of different culture media, temperatures and pH values on the accumulation of cell biomass was studied. Yoghurt samples were produced using a classical fermentation technology. Freeze-dried concentrated monoculture of L. gasseri 4/13 with over 1 × 1010 CFU/g viable cells was added as an adjunct culture together with a starter. The viable L. gasseri 4/13 cells remained above the critical value of 106 CFU/g during storage at 5 °C for the entire 20-day period. Organoleptic tests did not reveal any adverse change in the product taste and aroma of yoghurt samples at the 20th day. In conclusion, L. gasseri 4/13 was selected as having suitable probiotic and cultural characteristics for production of fermented milk products with high nutritional and biological value.

Differential modulation of innate immunity in vitro by probiotic strains of Lactobacillus gasseri

BackgroundProbiotics species appear to differentially regulate the intestinal immune response. Moreover, we have shown that different immune-modulatory abilities can be found among probiotic strains belonging to the same species. In this study, we further addressed this issue while studying L. gasseri, a species that induces relevant immune activities in human patients.ResultsWe determined the ability of two strains of L. gasseri, OLL2809 and L13-Ia, to alter cell surface antigen expression, cytokine production and nuclear erythroid 2-related factor 2 (Nrf2)-mediated cytoprotection in murine bone marrow-derived dendritic cells (DCs) and MODE-K cells, which represent an enterocyte model. Differential effects of L. gasseri strains were observed on the expression of surface markers in mature DCs; nevertheless, both strains dramatically induced production of IL-12, TNF-α and IL-10. Distinctive responses to OLL2809 and L13-Ia were also shown in MODE-K cells by analyzing the expression of MHC II molecules and the secretion of IL-6; however, both L. gasseri strains raised intracellular glutathione. Treatment of immature DCs with culture medium from MODE-K monolayers improved cytoprotection and modified the process of DC maturation by down-regulating the expression of co-stimulatory markers and by altering the cytokine profile. Notably, bacteria-conditioned MODE-K cell medium suppressed the expression of the examined cytokines, whereas cytoprotective defenses were significantly enhanced only in DCs exposed to OLL2809-conditioned medium. These effects were essentially mediated by secreted bacterial metabolites.ConclusionsWe have demonstrated that L. gasseri strains possess distinctive abilities to modulate in vitro DCs and enterocytes. In particular, our results highlight the potential of metabolites secreted by L. gasseri to influence enterocyte-DC crosstalk. Regulation of cellular mechanisms of innate immunity by selected probiotic strains may contribute to the beneficial effects of these bacteria in gut homeostasis.

Characterization of Lactobacillus gasseri isolates from a breast-fed infant

The potential health benefits of probiotic bacteria have led to the isolation of new microbial strains for incorporation into food products. However, newly isolated candidate probiotic organisms do not automatically share the “generally recognized as safe” (GRAS) status of traditional lactic acid bacteria (LAB). Before their introduction into food products, the safety of new isolates has to be evaluated. The objective of this study was to characterize LAB isolates from the stool of a newborn infant, and evaluate their safety and probiotic potential, in vitro. Thirty colonies were identified as Lactobacillus gasseri through sequencing of 16S rDNA. Pulsed Field Gel Electrophoresis using restriction enzymes SmaI and Apa I revealed that 29 of the L. gasseri were nearly identical, however one isolate exhibited a distinctive DNA fingerprint. All 30 L. gasseri were evaluated for resistance to antibiotics, bile tolerance, hemolytic activity and antagonism toward selected pathogens. All 30 strains harbored three plasmids, with one strain that showed strong tolerance to 0.5% of bile and harbored a unique fourth plasmid encoding a putative multidrug resistance transporter protein (LmrB). No hemolytic activity or antagonism, beyond acid inhibition was observed. Three selected strains UFVCC1083, 1091 and 1112 showed strong resistance to simulated small intestinal and gastric juices and adhered in vitro to mucin and two intestinal epithelial cell lines, Caco-2 and HT-29. This study identified and characterized recently isolated L. gasseri strains from faeces of a breast fed infant as potential probiotic candidates for use in the human milk banks in Brazil.

In vitro evaluation of Lactobacillus gasseri strains of infant origin on adhesion and aggregation of specific pathogens.

Numerous Lactobacillus species are members of the normal healthy human intestinal microbiota, and members of the Lactobacillus family predominate among the current marketed probiotic strains. Most of the current commercial probiotic strains have not been selected for specific applications but rather have been chosen based on their technological properties. Often the ability of such strains to temporarily colonize the gastrointestinal tract may be lacking, and the interactions with intestinal microbiota are few. Furthermore, the competitive exclusion properties of potential probiotic bacteria are strain specific and vary greatly. Thus, it is highly desirable that new candidate probiotic isolates originate from the healthy target population. In this study, seven newly isolated strains of Lactobacillus gasseri originating from feces of a healthy newborn child were evaluated for their ability to adhere to intestinal mucus, to autoaggregate and coaggregate with the model pathogens Cronobacter sakazakii (ATCC 29544) and Clostridium difficile (1296). All the bacterial strains, single or in combination, in viable and nonviable forms, were able to autoaggregate. The coaggregation with C. sakazakii or C. difficile was higher (P < 0.05) in nonviable than in the viable forms. Single L. gasseri strains showed similar adhesion abilities to intestinal colon mucus. The seven L. gasseri strains when combined were also able to significantly compete with, displace, and inhibit the adhesion of C. sakazakii and C. difficile in the mucus model. This study demonstrates that the studied L. gasseri strains fulfill the basic adhesion and aggregation properties for probiotics and could be considered for potential future use in children.

Strain-specific detection by pulsed-field gel electrophoresis of Lactobacillus gasseri TMC0356 in human feces after oral administration of these organisms

The present study aimed to develop an innovative, strain-specific means of identifying the probiotic Lactobacillus gasseri TMC0356 and to determine whether orally administered TMC0356 could be recovered from the human intestine. High molecular weight genomic DNA was isolated from TMC0356 and 14 reference strains of L. gasseri, including the type strain. The DNA samples were digested with the selected rare-cutting restriction endonucleases SmaI, SacII and ApaI and the resulting fragments separated by pulsed-field gel electrophoresis (PFGE) in a size range between 20 to 290 kb. TMC0356 could be distinguished from the other L. gasseri strains on the basis of the SmaI and SacII macrorestriction patterns. Furthermore, L. gasseri strains isolated from the feces of subjects who had ingested TMC0356 were identical to TMC0356 in the SmaI, SacII and ApaI macrorestriction fragments of digested DNA. These results suggest that PFGE of genomic DNA digested with SmaI, SacII, could be a practical means of identification of TMC0356. Furthermore, these results indicate that ingested TMC0356 can survive in, and colonize, the human intestine.

The probiotic Lactobacillus gasseri SBT2055 inhibits enlargement of visceral adipocytes and upregulation of serum soluble adhesion molecule (sICAM-1) in rats

The probiotic Lactobacillus gasseri strain SBT2055 (LG2055) was tested for its anti-obesity and anti-inflammatory effects to determine health benefits of consuming this organism and to explore possible relationships between those effects. Visceral adipocyte size was used as a measure of obesity, and the level of soluble intercellular adhesion molecule-1 (sICAM-1) in the blood as an inflammatory marker that is elevated in obesity. Comparisons were made among rats fed a 10% fat diet based on non-fermented milk, those fed a diet containing yoghurt fermented with conventional yoghurt bacteria, and those fed a diet containing yoghurt together with the probiotic LG2055. These experiments demonstrated that the yoghurt containing LG2055, but not the control yoghurt, inhibited the enlargement of visceral adipocytes and prevented upregulation of sICAM-1. These findings indicate the utility of specific probiotics and suggest a correlation between the anti-obesity effect and the anti-inflammatory action of LG2055.

Inulin and levan synthesis by probiotic Lactobacillus gasseri strains: characterization of three novel fructansucrase enzymes and their fructan products

Fructansucrase enzymes polymerize the fructose moiety of sucrose into levan or inulin fructans, with beta(2-6) and beta(2-1) linkages, respectively. Here, we report an evaluation of fructan synthesis in three Lactobacillus gasseri strains, identification of the fructansucrase-encoding genes and characterization of the recombinant proteins and fructan (oligosaccharide) products. High-performance anion-exchange chromatography and nuclear magnetic resonance analysis of the fructo-oligosaccharides (FOS) and polymers produced by the L. gasseri strains and the recombinant enzymes revealed that, in situ, L. gasseri strains DSM 20604 and 20077 synthesize inulin (and oligosaccharides) and levan products, respectively. L. gasseri DSM 20604 is only the second Lactobacillus strain shown to produce inulin polymer and FOS in situ, and is unique in its distribution of FOS synthesized, ranging from DP2 to DP13. The probiotic bacterium L. gasseri DSM 20243 did not produce any fructan, although we identified a fructansucrase-encoding gene in its genome sequence. Further studies showed that this L. gasseri DSM 20243 gene was prematurely terminated by a stop codon. Exchanging the stop codon for a glutamine codon resulted in a recombinant enzyme producing inulin and FOS. The three recombinant fructansucrase enzymes characterized from three different L. gasseri strains have very similar primary protein structures, yet synthesize different fructan products. An interesting feature of the L. gasseri strains is that they were unable to ferment raffinose, whereas their respective recombinant enzymes converted raffinose into fructan and FOS.

Lactobacillus spp. associated with early childhood caries

A group of 69 lactobacilli was isolated from caries lesions and root canals of early childhood caries (ECC) affected children treated in the Department of Pedodontics (Children's Teaching Hospital, Brno, Czech Republic). Biochemical and physiological properties of all strains were characterized by API 50 CH kit and conventional tube tests. The rep-PCR fingerprinting with the (GTG)(5) primer was used for genotypic grouping of the isolates. The (GTG)(5)-PCR fingerprinting grouped all analyzed strains into a few clusters in nearly full agreement with phenotype identification results and clarified the taxonomic position of 13 biochemically unidentified strains. In total, 20 strains of Lactobacillus fermentum, 17 L. rhamnosus, 14 L. casei/paracasei, 7 L. gasseri, 7 L. salivarius and 4 L. plantarum were identified. Mixtures of two or even three Lactobacillus spp. were isolated from a few root canal content samples. Results obtained by biotyping and (GTG)(5)-PCR were generally comparable except for L. gasseri strains that were not biochemically identified. The (GTG)(5)-PCR fingerprinting was shown to be quicker, easier to perform and more reliable than biotyping. Our results imply this molecular method as a good tool for screening and identification of Lactobacillus spp. inhabiting dental plaque.

Lactobacillusstrains isolated from the vaginal microbiota of healthy women inhibitPrevotella biviaandGardnerella vaginalisin coculture and cell culture

The purpose of this study was to investigate how human vaginal isolates of Lactobacillus acidophilus, Lactobacillus jensenii, Lactobacillus gasseri and Lactobacillus crispatus inhibit the vaginosis-associated pathogens Gardnerella vaginalis and Prevotella bivia. Results show that all the strains in coculture condition reduced the viability of G. vaginalis and P. bivia, but with differing degrees of efficacy. The treatment of G. vaginalis- and P. bivia-infected cultured human cervix epithelial HeLa cells with L. gasseri strain KS120.1 culture or cell-free culture supernatant (CFCS) results in the killing of the pathogens that are adhering to the cells. The mechanism of the killing activity is not attributable to low pH and the presence of lactic acid alone, but rather to the presence of hydrogen peroxide and proteolytic enzyme-resistant compound(s) present in the CFCSs. In addition, coculture of G. vaginalis or P. bivia with L. gasseri KS120.1 culture or KS120.1 bacteria results in inhibition of the adhesion of the pathogens onto HeLa cells.

Survival and in vivo adhesion of human isolates Lactobacillus gasseri LF221 and K7 in weaned piglets and their effects on coliforms, clostridia and lactobacilli viable counts in faeces and mucosa

In in vivo study on 24 weaned piglets (8 per group), the survival rates of human isolates Lactobacillus gasseri K7 and LF221 were quantified by selective enumeration on MRS agar with rifampicin, and the presence of both strains in intestinal mucosa was examined. Faeces from individual animals were analysed for the number (cfu/g) of coliforms, lactobacilli, clostridia and both of the two probiotic strains during 2-weeks probiotic application period (5 x 10(10) cfu of individual strain/day) and 1 week after the probiotic treatment was ceased. Samples of duodenum, jejunum and ileum of sacrificed animals (5th or 20th day) were also examined microbiologically. A great variability in the microflora of faeces and mucosa was observed even between equally treated animals. The survival of both Lb. gasseri strains was established by their detection in the faeces (2.5 x 10(5) to 3.3 x 10(5) cfu of K7 strain/g faeces; 4.5 x 10(5) to 5 x 10(5) cfu of LF221 strain/g). In two animals, the LF221 or K7 viable cells were found in the faeces 6 d after ceasing probiotic application. In both animals from the group fed with Lb. gasseri K7 that were sacrificed 5 d after weaning, the presence of K7 strain was found either in the mucosa of duodenum (140 cfu/10 cm2) and jejunum (170 cfu/10 cm2) or in the ileum (1600 cfu/10 cm2). LF221 cells were detected in the ileal mucosa of one piglet (820 cfu/10 cm2). The results demonstrated the capability of both tested strains of in vivo adhesion to intestinal mucosa and of temporary colonisation of the piglets' intestine.

Anaerobic Induction of Adherence to Laminin in Lactobacillus gasseri Strains by Contact with Solid Surface

The effect of growth conditions on adhesion was studied in six species belonging to Lactobacillus acidophilus homology groups. Namely, 17 strains including 6 fresh isolates of L. gasseri from human feces were assessed for their adherence to immobilized fibronectin, laminin, and type IV collagen. These extracellular matrix proteins were used as a model of damaged intestinal mucosa. When the bacteria were grown on MRS agar under anaerobic conditions, all eight L. gasseri strains and one L. johnsonii strain showed strong adhesiveness to laminin, but not when grown in static MRS broth. A similar pattern was observed in four L. gasseri strains in terms of adherence to fibronectin. No L. gasseri or L. johnsonii strains exhibited adhesion to type IV collagen under either growth condition. Adhesion of L. acidophilus, L. crispatus, L. amylovorus, and L. gallinarum was not affected by the growth conditions. Although protease treatment of L. gasseri cells abolished the adhesion, periodate oxidation of the cells increased it except in one strain. The adherence of L. gasseri cells was diminished by periodate and alpha-mannosidase treatments of immobilized laminin. The above results suggest that mannose-specific proteinaceous adhesion can be induced in L. gasseri by contact with a mucosal surface in the anaerobic intestinal lumen.

Marker-free chromosomal integration of the manganese superoxide dismutase gene ( sodA) from Streptococcus thermophilus into Lactobacillus gasseri

A strategy for functional gene replacement in the chromosome of Lactobacillus gasseri is described. The phospho-β-galactosidase II gene ( lacII) was functionally replaced by the manganese superoxide dismutase (MnSOD) gene ( sodA) from Streptococcus thermophilus, by adapting the insertional inactivation method described for lactobacilli [Russell, W.M. and Klaenhammer, T.R. 2001 Efficient system for directed integration into the Lactobacillus acidophilus and Lactobacillus gasseri chromosomes via homologous recombination. Appl. Environ. Microbiol. 67, 4361–4364]. L. gasseri carrying the heterologous sodA gene grew on lactose as efficiently as the wild-type parent. An active MnSOD was expressed in the transgenic strain, and the enzyme migrated on PAGE-SOD activity gels to the same position as that of MnSOD from S. thermophilus. The expression of MnSOD from a single copy of sodA integrated in the chromosome of L. gasseri provided enhanced tolerance to hydrogen peroxide, and extended the viability of carbon/energy starved cultures stored at 25 °C. This is the first report showing the successful utilization of the pORI plasmids system to generate marker-free gene integration in L. gasseri strains.

Oral administration of Lactobacillus and Bifidobacterium strains of intestinal and vaginal origin to healthy human females: Re-isolation from faeces and vagina

The healthy vaginal ecosystem is maintained by the interaction of the vaginal epithelium and the microflora, where lactobacilli and occasionally bifidobacteria play an important role. Imbalances in the dominance of lactobacilli may lead to bacterial vaginosis or yeast vaginitis. Probiotics may be a means to secure a favourable microbial balance in the vagina and, as the normal vaginal flora ascends from the rectal mucosa, a convenient form of administration may be via the oral gastrointestinal route. Ten healthy women of different ages were orally administered a blend of 12 selected Lactobacillus strains and three Bifidobacterium strains originating from the intestine and the vagina. The daily dose of each Lactobacillus strain was 1-3x10 9 CFU and 1x10 8 to 1x10 9 CFU for each Bifidobacterium strain, and the doses were given in an oatmilk/blueberry drink. The treatment went on for 10 days. Samples of faeces and vaginal fluid were taken immediately before the administration was started, after 10 days of administration and 7 days after the termination of the administration. Five strains were recovered from faeces and three from the vagina after treatment. The test strains could be found in 9 of the 10 volunteers. L. plantarum strains, originally isolated from intestinal mucosa, were recovered from the vagina of five subjects after treatment. The L. plantarum strains, together with one strain of L. crispatus and one L . gasseri strain, were recovered from faeces. Key words: Probiotics , Lactobacillus, vagina , oral administration , Bifidobacterium

Intracellular Enzyme Activities and Autolytic Properties of Lactobacillus Acidophilus and Lactobacillus Gasseri

A number of fermented milk products containing Lactobacillus acidophilus and Lactobacillus gasseri are now available as probiotic products. The proteolytic, lipolytic and autolytic properties of human-derived L. acidophilus (5 strains) and L. gasseri (7 strains) were evaluated, as these factors are closely related to cell viability and flavor development in the products. All L. gasseri strains showed higher intracellular protease activities than the L. acidophilus strains ; in contrast, lipase activities in L. gasseri were mostly lower than those of L. acidophilus. The many strains of L. gasseri were shown to have a greater tendency than L. acidophilus to autolyze in dispersed solution ; this liability was more distinct at lower pH values. These properties should be taken into account when bacteria are selected for production of probiotic products.

Selection of useful probiotic lactic acid bacteria from the Lactobacillus acidophilus group and their applications to functional foods

ABSTRACTIn the present review, a new mass screening system for selecting probiotic strains from Lactobacillus (L) acidophilus group lactic acid bacteria (LAB) with strong adhesion to the human intestinal tract is described. Characteristics of antimicrobial peptides (bacteriocin), lactose‐hydrolyzing enzymes and immunostimulative oligo DNA motifs in L. gasseri strains are also described. Finally, the use of L. acidophilus LAB, selected by our screening method, that have strong adhesion to the human colonic mucosa in functional yogurt products is described. Adhesiveness to the human intestine is one of the most important characteristics of probiotic LAB. A new screening system that involves a combination of three methods is proposed: rat colonic mucin (RCM)‐micro plate assay, Carnoy's histochemical staining method and carbohydrate probe binding assay. By using an RCM‐coated poly‐vinylidene‐diflouride membrane that mimics the human colonic mucous layer, a new lectin was isolated and its structure was clarified by gene cloning. Furthermore, the structures and functions of a new cyclic bacteriocin (gassericin A), new lactose‐hydrolyzing enzymes, and new immunostimulating oligo DNA motifs from Lactobacillus gasseri (B1 subgroup) were clarified. A new functional yogurt ‘Fit down’ is proposed, that is fermented by an adhesive strain of L. acidophilus LA67 selected by our screening and contains antihypertensive peptides derived from whey proteins by protease digestion. In the future, superior functional foods containing more effective probiotic LAB are expected to be developed by the use of the proposed mass screening system.

Antibacterial activity associated with Lactobacillus gasseri ATCC 9857from the human female genitourinary tract

The 10-fold concentrated spent MRS culture cell-free supernatant concentrate [(cCFS)] of the human female genitourinary tract isolate Lactobacillus gasseri ATCC 9857 was shown to exhibit antibacterial activity towards gram-positive sporogenous and asporogenous fermentative eubacteria in liquid and on solid media under conditions that eliminated the activity of lactic acid (β-glycerophosphate) and hydrogen peroxide (catalase). The antibacterial activity of the cCFS was characterized by automated turbidometry (Bioscreen™) and non-linear regression analysis (Gompertz model) using MRS broth cultures of the indicator strain L. acidophilus ATCC 11975. It exhibited a bactericidal mode of action, sensitivity to trypsin and proteinase K, partial sensitivity to pepsin and pronase E, partial heat stability at 121 °C for 15 min, and retained significantly more activity following exposure to pH 3.0 and 5.0 compared with pH 7.2 and 9.0. The inhibitory spectrum included a wide range of Lactobacillus species, Bifidobacterium bifidum, B. infantis and B. catenulatum, Lactococcus cremoris, Leuconostoc cremoris, Pediococcus pentosaceus, Bacillus cereus, Clostridium tyrobutyricum, C. pasteurianum, C. sporogenes, Staphylococcus carnosus, and Enterococcus faecalis. Although partial inhibition of Escherichia coli ATCC 25922 by cCFS was observed in liquid medium, inhibition of freshly isolated human uropathogenic E. coli strains could not be demonstrated on TSB agar plates by agar well diffusion. Following partial resolution by gel permeation FPLC on Superose-12, the fractionated cCFS was shown to comprise at least two inhibitory peptides (3.05 and 5.27 kDa) as well as aggregated inhibitory peptide material (21.65, 41.50, 81.20, and 120.90 kDa). The 3.05 kDa peptide, designated Gassericin D, inhibited L. acidophilus strains ATCC 11975 and ACA-DC 241. The 5.27 kDa peptide, designated Gassericin C, inhibited L. gasseri strain UCSC LF221Snb and En. faecalis DPC 3319. The aggregated 21.65 kDa peptide material strongly inhibited L. acidophilus ATCC 11975 and weakly inhibited Listeria inocua DPC 3306. The aggregated 41.50 kDa peptide material strongly inhibited Ba. cereus DPC 3316 and weakly inhibited L. acidophilus ACA-DC 241. The ability of L. gasseri ATCC 9857 to produce bacteriocin-like activity may be of importance in the biopreservation of nutraceuticals and in the management of female genitourinary and gastrointestinal tract infections involving En. faecalis.

Isolation and characterization of a new bacteriocin from Lactobacillus gasseri KT7.

A bacteriocin-producing Lactobacillus gasseri strain, KT7, was isolated from infant faeces. The supernatant fluid showed inhibitory activity not only against some lactic acid bacteria but also, against some pathogenic and food-spoilage species, including Clostridium, Listeria and Enterococcus. An antimicrobial peptide designated gassericin KT7 was isolated from Lactobacillus gasseri KT7. It was purified to homogeneity by a single four-step procedure: a crude supernatant fluid obtained from early stationary-phase culture in MRS medium was subjected to ammonium sulphate fractionation, CM-Sephadex cation-exchange chromatography, Phenyl-Sepharose hydrophobic chromatography and reverse-phase HPLC chromatography. Gassericin KT7 was sensitive to proteolytic enzymes, resistant to heat, active over a wide range of pH, and migrated as a 4.5-5.0 kDa peptide on SDS-PAGE. The bacteriocin was produced constitutively during exponential growth. It was bactericidal to sensitive cells and the bactericidal effect was not produced by cell lysis. The amino acid composition of the bacteriocin was determined and no modified amino acid was found among the residues identified.

Viability of Lactobacillus gasseri and Its Cholesterol-Binding and Antimutagenic Activities During Subsequent Refrigerated Storage in Nonfermented Milk

The effect storage at 4°C on the viability of Lactobacillus gasseri and its sodium taurocholate-deconjugating and cholesterol-binding abilities as well as desmutagenic activity was investigated. Unfermented milks containing L. gasseri strains SBT0274 and SBT0270 at 109 cfu/ml were prepared using 10% skim milk. Total and bile-tolerant lactobacilli for strains SBT0274 and SBT0270 generally decreased after 14 d of storage at 4°C; however, viable cells of these strains were still at 108 cfu/ml after 28 d of storage. The amounts of cholic acid released and of cholesterol bound by strains SBT0274 and SBT0270 declined over time, especially at 21 d of storage. Antimutagenic activity of unfermented milk made from both strains was attributed to the bacterial cells, and the activity was stable during storage at 4°C for 28 d.

Bile tolerance, taurocholate deconjugation, and binding of cholesterol by Lactobacillus gasseri strains.

Bile tolerance, deconjugation of sodium taurocholate, and the cholesterol-binding ability of 28 strains of Lactobacillus gasseri were examined. There was significant variation among strains in growth in media containing bile and also variation in the ability to bind cholesterol. Cultures grown for 12 h at 37 degrees C bound significantly more cholesterol than did cells from a 48-h incubation. Variation among strains in the ability to deconjugate sodium taurocholate was not significantly different. Maximal deconjugation of sodium taurocholate was achieved with the cells during the stationary phase of growth (12 h). Statistical analysis showed no significant correlation between bile tolerance and sodium taurocholate deconjugation, bile tolerance and cholesterol-binding ability, or sodium taurocholate deconjugation and cholesterol-binding ability.