• Journal of Microbiology and Biotechnology
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• v.18 no.12
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• pp.1953-1957
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• 2008

Recombinant Lactobacillus strains have been constructed using gene splicing by overlap extension (SOE). Primers were designed of which one end of an amplified product contained complementary sequences for an end of other amplified fragment. For efficient matching, we used an asymmetric PCR step that was effective at generating an excess of strands that would anneal in the final PCR. CP12, a recombinant fragment consisting of the integrase gene and attachment site of the bacteriophage A2, was constructed and inserted into the genome of Lactobacillus casei ATCC 393, yielding Lb. casei ATCC 393::XCP12. Another recombinant Lb. casei strain was constructed, where the egfp gene was a part of the construction. The EGFP produced from Lb. casei ATCC 393::XCEGFP14 was detected by Western blot hybridization. This simple and widely applicable approach has significant advantages over standard recombinant DNA techniques for Lactobacillus species.

• Journal of Animal Science and Technology
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• v.48 no.5
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• pp.719-726
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• 2006

Bovine whey protein are rich in cysteine, which is the rate limiting amino acid for synthesis of antioxidant glutathione(GSH). Some strains of Lactobacillus caseihas been reported to contain high level of GSH in cell extracts. The objective ofthis study was to determine whether enzymatically hydrolyzed whey protein isolate(WPI) and cell extract of Lb. casei HY2782 could increase intracellular GSH concentrations and protect against oxidant induced cell death in human prostate epithelial cell line (designated as RWPE1, and PC3MMM2 cells). Treatment of RWPE1 cellsandPC3MMM2 cells with hydrolyzed WPI (500g/ml) significantly increased GSH by28.2% and38.4% respectively. Compared with control cells receiving no hydrolyzed WPI(P<0.05). hydrolyzed WPI and Lb casei HY2782 cell extracts significantly protected RWPE1 and PC3MMM2 cellsfrom oxidant induced cell death compared with controls receiving no WPI. DNA damage associated with oxidant treatment was demonstrated by single cell gel (SCG) electrophoresis.

• Preventive Nutrition and Food Science
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• v.14 no.2
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• pp.162-167
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• 2009

Various fermented foods were screened in search of food-grade bacteria that produce bacteriocins active against Gram-negative pathogens. An isolate from a mold-ripened cheese presented antibacterial activity against Gram-positive and Gram-negative bacteria. The most active isolate was identified as Lactobacillus casei by a biochemical method, ribotyping, and membrane lipid analysis, and was designated as OSY-LB6A. The cell extracts of the isolate showed inhibition against Escherichia coli p220, E. coli O157, Salmonella enerica serovar Enteritidis, Salmonella Typhimurium, and Listeria monocytogenes. The antibacterial nature of the cell extract from the isolate was confirmed by eliminating the inhibitory effects of acid, hydrogen peroxide, and lytic bacteriophages. The culture supernatant and cell extract retained antibacterial activity after heating at $60{\sim}100^{\circ}C$ for $10{\sim}20$ min. The activity of the cell extract from Lb. casei was eliminated by pronase and lipase. Finally, the cell extract showed a bactericidal mode of action against E. coli in phosphate buffer solution, but it was bacteriostatic in broth medium and food extracts.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.2
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• pp.255-258
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• 2005

Several commercial yogurt starter cultures (L. acidophilus LA5, L. casei 01, L. acidophilus LA100, L. bulgaricus LB207 and L. rhamnosus GG744) were investigated for antioxidant activity by using in vitro assays. From the results of the present work, all strains tested showed varying degrees of antioxidant activity. L. bulgaricus LB207 showed the highest antioxidant activity, with a linoleic acid peroxidation inhibition of 81.3%. Hydroxy radical scavenging activity, ferrous iron chelating activity, reducing power and superoxide dismutase (SOD) activity were also studied. L. bulgaricus LB207 showed the highest hydroxy radical scavenging activity and L. casei 01 showed the highest chelating activity. L. bulgaricus LB207 and L. acidophilus LA100 showed good reducing power. All the strains in this study showed low SOD activity. The results of the present work suggest that antioxidant activity of L. bulgaricus LB207 due to its strong hydroxy radical scavenging activity and reducing power.

• Food Science and Biotechnology
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• v.17 no.6
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• pp.1221-1227
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• 2008

The application of the cellulase gene (celA) as a selection marker of food-grade integration system was investigated in Lactobacillus (Lb.) casei, Lactococcus lactis, and Leuconostoc (Leu.) mesenteroides. The 6.0-kb vector pOC13 containing celA from Clostridium thermocellum with an integrase gene and a phage attachment site originating from bacteriophage A2 was used for site-specific recombination into chromosomal DNA of lactic acid bacteria (LAB). pOC13 was also equipped with a broad host range plus replication origin from the lactococcal plasmid pWV01, and a controllable promoter of nisA ($P_{nisA}$) for the production of foreign proteins. pOC13 was integrated successfully into Lb. casei EM116, and pOC13 integrants were easily detectable by the formation of halo zone on plates containing cellulose. Recombinant Lb. casei EM 116::pOC13 maintained these traits in the absence of selection pressure during 100 generations. pOC13 was integrated into the chromosome of L. lactis and Leu. mesenteroides, and celA acted as an efficient selection marker. These results show that celA can be used as a food-grade selection marker, and that the new integrative vector could be used for the production of foreign proteins in LAB.

• Korean Journal of Food Science and Technology
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• v.47 no.2
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• pp.204-210
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• 2015

Makgeolli is made from rice or flour, yeast, and nuruk, a fermentation starter. The flavor of makgeolli is affected by sugars, amino acids, organic acids and volatile flavor compounds produced by various microorganisms. In this study, lactic acid bacteria (LAB) were isolated from unsterilized makgeolli samples collected from several provinces in Korea, and then later identified. Under anaerobic conditions, LAB density ranged from $5.0{\times}10^6$ to $1.5{\times}10^8CFU/mL$; yeast density ranged from $2.5{\times}10^7$ to $1.5{\times}10^8CFU/mL$. Of the LAB isolated from makgeolli, 1,126 were analyzed using restriction fragment length polymorphism (PCR-RFLP) analysis of 16S rRNA, which allowed for classification into five groups. Of the 1,126 LABs tested, 130 produced ${\gamma}$-aminobutyric acid (GABA).

• Journal of Food Hygiene and Safety
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• v.6 no.3
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• pp.119-126
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• 1991

ABSTRACT - This study was performed to investigate the possible effect of Lactobacillus spp. on the growth and citrinin production by Penicillium citrinum. Lactobacillus bulgaricus and Lactobacillus casei were grown with Pen. citrinum in modified APT broth containing 7% of glucose and incubated at $30^{\circ}C$ for 15 days. Four inoculation procedures were used; (a) Lactobacillus spp. and Pen. citrinum were grown alone(Pc, Lb, and Lc), (b) both organisms were added simultaneously(ST; Pc+Lb and Pc+Lc), (c) Lactobacillus spp. was grown 3 days, then conidia of Pen. citrinum were added(LbPc and LcPc), and (d) Pen. citrinum was grown 3 days, then Lactbacillus spp. was added (PcLb and PcLc). At 0, 3, 6, 9, 12, 15 days of incubation, the growth of each organism, pH and total acidity of broth, and content of citrinin were determined. Lactobacillus spp. and Pen. citrinum, when grown associatively, influenced the growth of each other. It was observed that slower growth of Pen. citrinum when in the presence of Lactobacillus spp. than when the mold grew alone. Production of citrinin by Pen. citrinum was markedly less in the mixed culture. No apparent growth and toxin production was observed when the Lactobacillus spp. was grown 3 days, then conidia of Pen. citrinum were added(LbPc and LcPc). The above results indicate that another microorganism or competing microflora in the culture can affect the behavior of Pen. citrinum.trinum.

• The Korean Journal of Physiology and Pharmacology
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• v.20 no.6
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• pp.621-628
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• 2016

Hypertension can be caused by various factors while the predominant causes include increase in body fluid volume and resistance in the circulatory system that elevate the blood pressure. Consumption of probiotics has been proven to attenuate hypertension; however, the effect is much strain-dependent. In this study, a newly isolated Lactobacillus casei (Lb. casei ) strain C1 was investigated for its antihypertensive properties in spontaneously hypertensive rats (SHR). Lactic acid bacteria (LAB) suspension of 11 log colony-forming unit (CFU) was given to SHR (SHR+LAB, n=8), and phosphate buffer saline (PBS) was given as a control in SHR (SHR, n=8) and in Wistar rats as sham (WIS, n=8). The treatment was given via oral gavage for 8 weeks. The results showed that the weekly systolic blood pressure (SBP), mean arterial pressure (MAP), diastolic blood pressure (DBP) and aortic reactivity function were remarkably improved after 8 weeks of bacterial administration in SHR+LAB. These effects were mostly attributed by restoration of wall tension and tensile stress following the bacterial treatment. Although not statistically significant, the level of malondialdehye (MDA) in SHR+LAB serum was found declining. Increased levels of glutathione (GSH) and nitric oxide (NO) in SHR+LAB serum suggested that the bacterium exerted vascular protection through antioxidative functions and relatively high NO level that induced vasodilation. Collectively, Lb. casei strain C1 is a promising alternative for hypertension improvement.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.10
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• pp.1476-1480
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• 2016

Anti-microbial effects of the medium extracts from three different lactic acid bacteria (LB1, Lactobacillus acidophilus; LB2, Lactobacillus casei; LB3, Lactobacillus sicerae) were investigated. Three different extracts of lactic acid bacteria media (ELAM) did not show significant changes in pork loin quality after 3 and 14 days of cold storage such as general contents, colors, pH, and TBARS. To determine anti-bacterial activity of three ELAM, three pathogenic bacteria (Bacillus cereus, Bacillus subtilis, and Staphylococcus aureus) were obtained and incubated with ELAM-absorbed paper discs. ELAM of LB2 and LB3 showed significantly larger bacterial growth inhibitory zones compared with that of LB1 (P<0.05). When the pathogenic bacteria inoculated in pork loin with three ELAM, total microbial contents of pork loin treated with ELAM of LB3 after 14 days of cold storage showed significantly lower microbial contents compared to those of control, LB1 and LB2 (P<0.05). In conclusion, ELAM of LB3 derived from L. sicerae had the most effective pathogenic bacteria inhibitory activity on agar and pork loin. This is the first result to report the antibacterial effect of L. sicerae. If the safety and toxicity characteristics of L. sicerae are further investigated, this new lactic acid bacterium would have potential as an effective and nature-friendly food preserving agent.

• Journal of Life Science
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• v.23 no.1
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• pp.44-54
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• 2013

To determine the potential function of Rhus javanica in Korean medicine, it was fermented with each strain of Lactobacillus spp. Each strain of Lactobacillus spp. was inoculated in lactobacilli MRS broth, and 5 mg/ml of methanol extract of Rhus javanica was added. In mouse hippocampal HT22 cells, ethyl acetate extract of R. javanica fermented with L. brevis KCTC 3498 induced heme oxygenase-1 expression and showed a significant cytoprotective effect on glutamate-induced oxidative damage. The cytoprotective effect was related to the transcription of the nuclear factor E2-related factor2 (Nrf2), which is responsible for the induction of heme oxygenase-1 within the nucleus. The antimicrobial, antioxidant, and heme oxygenase-1 expression activities of fermented R. javanica were measured after extraction with ethyl acetate. R. javanica fermented with L. plantarum subsp. plantarum KCTC 3108, L. fermentum KCTC 3112, and L. brevis KCTC 3498 had higher antioxidant activity than nonfermented R. javanica. The fermented R. javanica with L. plantarum subsp. plantarum KCTC 3108, L. casei KCTC 3109 after ethyl acetate extraction showed antibacterial activity against Bacillus subtilis PCI 219, Escherichia coli KCTC 1682, Shigella flexneri KCTC 2517, Vibrio parahaemolyticus KCTC 7471, and Pseudomonas aeruginosa KCTC 2004. An ethyl acetate extract of the fermented R. javanica with Lactobacillus brevis KCTC 3498 exhibited stronger antibacterial activity than a nonfermented one against strains of B. subtilis PCI 219, E. coli KCTC 1682, S. flexneri KCTC 2517, and V. parahaemolyticus KCTC 7471.