• KSBB Journal
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• v.15 no.4
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• pp.411-413
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• 2000

A method is described for the rapid and simple isolation of genomic DNA from 3 mL culture of Lactobacillus crispatus KLB46 The isolated DNA using this method was shown to be an excellent substrate for restriction endonclease digestion and PCR. The method is expected to be used in gentic manipulation of L. crispatus KLB46.

• Korean Journal of Poultry Science
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• v.27 no.1
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• pp.37-41
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• 2000

This study was carried out to examine the effects of Lactobacillus spp. on performance and cecal microflora in broiler chicks. Eight diets consisting of basal diet(C), C+Lactobacillus crispatus BC7 (T2), C+Lactobacillus reuteri BC9 (T3), C+L.reuteri BC5+L.crispatus BC9 (T4), C+L. reuteri BC5 (T1)+L.reuteri BC9 (T5), C+L.crispatus BC7+L.reuteri BC9 (T6) and C+L.reuteri BC5+L.crispatus BC7+L.reuteri BC9 (T7), were fed to Ross male broiler chicks for 5 weeks. The level of supplemented Lactobacillus spp. was 107 cfu/g diet. Body weight and feed intake were measured every week, and cecal microfla was counted at 1 and 5 weeks for Lactobacillus and yeast. Body weight increased signigicantly in supplemental Lactobacillus treatments from 2 weeks of age (p<0.05). Chicks in T2, T5 and T5 of treatments were heavier than those of other treatments at 5 weeks of age(p<0.05). Viability was not different significantly. Feed intake and feed conversion also were not different, although feed conversion improved slightly in supplemental Lactobacillus treatments. Lactobacillus spp. of cecal content was increased in supplemental Lactobacillus treatments at 1 week, but no significance was found. The number of yeast in cecum was not different from that of supplemental Lactobacillus. In conclusion, Lactobacillus, L.crispatus BC7 and L.reuteri BC9 from broiler cecum could contribute to the increase in body weight with supplemented mono-or mixing of di-lactobacilli. The results indicate that Lactobacillus feeding can benefit to intestinal lactobacillus at early growing broiler.

• Journal of Microbiology
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• v.39 no.2
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• pp.146-148
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• 2001

Lactobacillus crispatus ATCC 33820 and L. gasseri ATCC 33323 were grown in MRS broth (pH 6.5) at 37$^{\circ}C$ for 24 h and the antibacterial activities of cell free culture supernatants were determined by the agar well diffusion method. The culture supernatants were inhibitory to Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis, Pediococcus acidilacticii, and Lactobacillus helveticus. The supernatants did not show any lysozyme activity. Addition of catalase did not affect the antibacterial activities of the supernatants. The antibacterial substances were heat stable (100$^{\circ}C$ for 60 min) and sensitive to proteases.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.2
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• pp.128-132
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• 2001

Bacterial vaginosis can be treated by restoring the normal vaginal flora using lactobacilli. Lactobacillus crispatus KLB46 that was isolated from the human vagina has a string antimicrobial activity and was grown in a batch and in a continuous fermentor. During batch cultivation, the maximum specific growth rate of L. crispatus KLB 46 was 0.63h(sup)-1 and the highest viable cell count (1.9$\times$10(sup)9 CFU/mL) was obtained at pH 5.5. L. crispatus KLB 46 did not grow well at either pH 3.5 or 7.5. During continuous cultivation, the highest viable cell count (1.53$\times$10(sup)9 CFU/mL) was obtained at a dilution rate of 0.32h(sup)-1, and was 7.33$\times$10(sup)11 CFU L(sup)-1 h(sup)-1, that is approximately 5 times higher than that obtained from batch culture.

• KSBB Journal
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• v.26 no.6
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• pp.529-532
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• 2011

Whey based medium was optimized for the production of viable Lactobacillus crispatus KLB 46 isolated from the vagina of Korean women. Among the various nitrogen sources such as yeast extract, beef extract, and proteose peptone no. 3 supplemented to whey, beef extract showed the highest viable cell production. The addition of Tween 80 to the whey based medium increased viable cell concentration. As beef extract supplementation is not economically attractive, corn steep liquor was added as a supplementary nitrogen sources. When corn steep liquor was supplied with beef extract with the ratio 5 : 1, the viable cell count was $3.11{\times}10^9$ CFU/mL. Also, the addition of mineral salts containing sodium acetate (5 g/L), potassium phosphate dibasic (2 g/L), magnesium sulfate (0.1 g/L) and manganese sulfate (0.05 g/L) to the whey medium increased viable cell count further ($5.00{\times}10^9$ CFU/mL).

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1176-1183
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• 2009

Lactobacillus crispatus (L. crispatus) ZJ001 is highly adhesive to epithelial cells and expresses S-layer proteins. In this study, S-S-layer layer genes were sequenced and expressed in E. coli to characterize the function of proteins with this particular strain. L. crispatus ZJ001 harbored two S-layer genes slpA and slpB, and only slpA gene was expressed in the bacterium, as revealed by RT-PCR and immunoassays. The mature SlpA showed 47% amino acid sequence identity to SlpB. The SlpA and SlpB of L. crispatus ZJ001 were highly homologous at the C-terminal region to other Lactobacillus S-layer proteins, but were substantially variable at N-terminal and middle regions. Electron microscopic analysis indicated that His-slpA expressed in E. coli was able to form a sheet-like structure similar to the natural S-layer, but His-slpB formed as disc-like structures. In the cell binding experiments, HeLa cells were able to bind to both recombinant His-slpA and His-slpB proteins to the extent similar to the natural S-layer. The cell binding domains remain mostly in the N-terminal regions in SlpA and SlpB, as shown by high binding of truncated peptides SlpA2-228 and SlpB2-249. Our results indicated that SlpA was active and high binding to HeLa cells, and that the slpA gene could be targeted to display foreign proteins on the bacterial surface of ZJ001 as a potential mucosal vaccine vector.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.10
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• pp.4255-4259
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• 2014

Lactobacilli are probiotics shown to have antitumor activities. In addition, they can regulate gene expression through epigenetic mechanisms. In this study, we aimed to assess anti tumor activities of Lactobacillus acidophilus and Lactobacillus crispatus on the MDA-MB-231 breast cancer cell line. The effects of culture supernatants were determined by MTT [3-(4,5-dimethylthiazol-2-y-2,5-diphenyltetrazolium bromide] assay. Changes in expression of 5 cancer-testis antigens (CTAs), namely AKAP4, ODF4, PIWIL2, RHOXF2 and TSGA10, were analyzed by quantitative real time RT-PCR. The culture supernatants of the 2 lactobacilli inhibited MDA-MB-231 cell proliferation. In addition, transcriptional activity of all mentioned CTAs except AKAP4 was significantly decreased after 24 hour treatment with culture supernatants. This study shows that Lactobacillus acidophilus and Lactobacillus crispatus have antiproliferative activity against MDA-MB-231 cells. In addition, these lactobacilli could decrease transcriptional activity of 4 CTAs. Previous studies have shown that expression of CTAs is epigenetically regulated, so it is possible that lactobacilli cause this expression downregulation through epigenetic mechanisms. As expression of CTAs in cancers is usually associated with higher grades and poor prognosis, downregulation of their expression by lactobacilli may have clinical implications.

• KSBB Journal
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• v.16 no.6
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• pp.626-631
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• 2001

Lactobacilli have been considered to play important roles in the health of human vagina. They secrete inhibitory substances to prevent vaginal infection by pathogenic organisms. In a previous study, we have isolated several lactobacilli from Korean woman and one of them (KLB46) was selected and indentified as Lactobacillu crispatus which showed high antimicrobial activity. In this study. cold adaptation prior to subsequent stresses exposure was examined whether L. crispatus KLB46 maintain the viability better than the non-adapted calls under stresses. For pharmaceutical formulation, the lyophilization process is required where stresses such as freezing/thawing and dehydration are routinely applied. Formulated L. crispatus KLB46 can be used for ecological treatment of bacterial vaginosis. The response of cold-adapted cells to other environmental stresses such as acid, heat, ethanol, NaCl, and H$_2$O$_2$ was also examined. The results showed that cold-adapted cells maintained higher survival rate compared with the non-adapted cells (freezing-thawing. 3-folds; dehydration: 3-folds; acid, 3-folds; heat, 10-folds). However, we did net observe any positive effect of cold adaptation on other stresses such as ethanol, NaCl and H$_2$O$_2$. When chloramphenicol was added during cold adaptation, adaptation effect was abolished. This confirms that de novo protein synthesis is necessary during the adaptation process. Moreover, we have identified cold shock protein homolog that codes for a major cold shock protein by PCR amplification using degenerate primers.

• KSBB Journal
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• v.29 no.2
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• pp.81-86
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• 2014

Lactobacilli, the dominant species of microorganisms in the vaginal flora of healthy women, play important roles to prevent bacterial vaginosis and other sexually transmitted diseases. In this study, we carried out studies on stress adaptation prior to various stress treatment. We found that heat or salt adapted KLB46 showed higher cell viability than non adapted upon heat stress at $60^{\circ}C$ for 20 min. When chloramphenicol was added during the adaptation process, heat tolerance was abolished. This result suggested that de novo protein synthesis was essential during adaptation.

• Journal of Animal Science and Technology
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• v.50 no.4
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• pp.509-518
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• 2008

This experiment was conducted to investigate enzyme activity, antimicrobial activity, and antibiotics susceptibility of Lactobacilli strain(Lactobacillus reuteri BLA5, Lactobacillus crispatus BLA7, Lactobacillus reuteri BLA9, Lactobacillus amylovorus LLA7, Lactobacillus crispatus LLA9, Lactobacillus vaginalis LLA11) isolated from chicken ceca and were selected by organic acid synthesis, acid tolerance, bile salt tolerance. The enzymes activities were different among strains of Lactobacilli. The amylase activity and lipase activity of Lactobacillus were high but cellulase activity and protease activity of that were low. Lactobacillus culture showed high antimicrobial activity against E. coli but low antimicrobial activity against Salmonella. The inhibitory factor of Lactobacilli isolated from chickens’ cecum on E. coli was low pH by organic acid. All of Lactobacillus isolated from chicken’s cecum were susceptible to ampicillin and amoxicillin but weren’t susceptible at the optimum level of feed additive antibiotics(virginiamycin and salinomycin).