• Preventive Nutrition and Food Science
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• v.9 no.4
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• pp.324-329
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• 2004

In order to investigate the molecular mechanism of $\gamma$-aminobutyric acid (GABA) production in lactic acid bacteria, we cloned a glutamate decarboxylase (GAD) gene from Lactobacillus plantarum using polymerase chain reaction (PCR). One PCR product DNA was obtained and inserted into a TA cloning vector with a T7 promoter. The recombinant plasmid was used to transform E. coli. The insertion of the product was con­firmed by EcoRI digestion of the plasmid purified from the transformed E. coli. Nucleotide sequence analysis showed that the insert is a full-length Lactobacillus plantarum GAD and that the sequence is $100\%$ and $72\%$ identical to the regions of Lactobacillus plantarum GAD and Lactococcus lactis GAD sequences deposited in GenBank, accession nos: NP786643 and NP267446, respectively. The amino acid sequence deduced from the cloned Lactobacillus plantarum GAD gene showed $100\%$ and $68\%$ identities to the GAD sequences deduced from the genes of the NP786643 and NP267446, respectively. To express the GAD protein in E. coli, an expression vector with the GAD gene (pkk/GAD) was constructed and used to transform the UT481 E. coli strain and the expression was confirmed by analyzing the enzyme activity. The Lactobacillus plantarum GAD gene obtained may facilitate the study of the molecular mechanisms regulating GABA metabolism in lactic acid bacteria.

• Journal of Animal Science and Technology
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• v.50 no.6
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• pp.807-818
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• 2008

This study was conducted to prove the optimum feeding method of Lactobacillus in broiler chickens by investigating the intestinal viability of ingested Lactobacillus and the effect of feeding levels and frequency of Lactobacillus on growth performance in broiler chickens. In experiment 1, A total of one hundred, 5 weeks old male broiler chickens(Abor Acre) were fed Lactobacillus reuteri avibro2 expressed green fluorescent protein(GFP) at 104cfu/g diet to investigate the retention time of ingested Lactobacillus in the intestine for 1 day. The percentage of Lactobacillus expressed GFP in intestinal contents was 26% at 1 day after fed Lactobacillus expressed GFP. The percentage of Lactobacillus expressed GFP in intestinal contents was decreased in length of time. In experiment 2, A total of four hundred eighty, 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each to prove the optimum feeding level of Lactobacillus. The treatments were control(free antibiotics), Lactobacillus reuteri avibro2 5.0×10cfu/mL, 5.0×103cfu/mL, and 5.0×105cfu/mL. The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). Feed conversion ratio was not significantly difference among the groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). Protein and fat digestibility in Lactobacillus 5.0×103cfu/mL and 5.0×105cfu/mL treated groups were significantly improved(P<0.05). No significant differences were observed on the availability of dry matter and crude ash in Lactobacillus treatments compared to those of control. In experiment 3, A total of six hundred 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each and were fed Lactobacillus reuteri avibro2 at intervals of 1, 2, 3, and 5 day for five weeks. Feeding level of Lactobacillus was 5.0×103cfu/mL The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). The final body weight and body weight gain were significantly increased, when Lactobacillus was fed at intervals of 1 days, or 2 days. There were no significant differences in feed intake and feed conversion ratio among the all groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). No significant differences were observed on the number of coliform bacteria and Salmonella of ileum and cecum. Consequently, supplemental Lactobacillus influenced positive effects on the growth performance, nutrient availability and intestinal microflora. The optimum feeding level of Lactobacillus was 5.0×103cfu/mL, and the constant feeding of Lactobacillus was effective.

• The Korean Journal of Food And Nutrition
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• v.10 no.3
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• pp.350-359
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• 1997

The changes in pH, acid contents and microbial counts were investigated during fermentation of Baikkimchi, a kind of Kimchi without red pepper, and the major microbial groups were also isolated and identified. Immediately after the preparation of Baikkimchi(pH 6.15, acid contents 0.03%), its major microbial group was Gram negative rods, and was composed of Pseudomonas(55%), Enterobacter(40%) and Erwinia(5%). After 2 days of fermentation at 15$^{\circ}C$, the most predominant microbial group was changed to lactic acid bacteria. Lactic acid bacteria showed 1st, 2nd and 3rd stationary phase on its growth curve in 4, 12 and 50 days of fermentation, respectively. At the 2nd stationary phase of lactic acid bacteria(pH 3.51, acid contents 0.59%), the group was composed of Lactobacillus bavaricus(55%), Leuconostoc mesenteroides subsp. mesenteroides(42.5%) and Leuconostoc paramesenteroides(2.5%), while at the 3rd stationary phase(pH 3.40, acid contents 1.10%), that was Lactobacillus plantarum(65%) and Lactobacillus brevis(35%). The physiological and biochemical characteristics identified as Leuconostoc mesenteroides subsp. mesenteroides, Leuconostoc paramesenteroides, Lactobacillus plantarum and Lactobacillus brevis showed good agreement with the current taxonomic system, but those identified as Lactobacillus bavaricus showed some disagreements. The number of yeast was decreased wit the increase in the number of lactic acid bacteria. Yeast showed stationary phase in 30 days between the 2nd and 3rd stationary phase of lactic acid bacteria, and the group was composed of only gunus Saccharomyces.

• Journal of Animal Science and Technology
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• v.45 no.3
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• pp.473-482
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• 2003

In vivo protective and in vitro inhibitory activities of Lactobacillus casei YIT 9018. against typical enteritis causing Salmonella enteritidis KU101 and IgA level after challenge have been determined. In order to identify the strains of lactobacilli the sequences of 16S-23S rRNA intergenic spacer region were determined. All the test strains of Lactobacillus spp. inhibited Salmonella enteritidis, the intensity varied depending upon the species of lactobacilli. Effects on the survival rate of the mouse after challenge with Salmonella enteritidis KU101 on feeding Lactobacillus spp. have shown the highest survival rate in L. helveticus CU 631 followed by L. casei YIT 9018 and L. johnsonii C-4 and the lowest in control mice. The higher level of total Ig A concentration in the intestinal fluid of lactobacilli fed mice than control mice was observed. The sequences of 16S-23S rRNA intergenic spacer region of seven strains of Lactobacillus casei could be utilized as a strain identification, those sequences showed some degree of difference in homology.

• KSBB Journal
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• v.24 no.2
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• pp.182-188
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• 2009

Lactobacillus spp., primary members of probiotics, have significant benefits for health and well-being of human. In this study Lactobacillus strains representing six species (L. paracasei KLB58, L. fermentum MS79 and KLB282, L. plantarum KLB213, L. gasseri KLB238, and L. reuteri KLB270) isolated from Korean adults were electrotransformed with plasmid pNCKH104. To determine optimal electrotransformation conditions, various conditions including cell wall weakening agent, electroporation buffer, electric field strength and time constant were tested for each strain. Overall, high transformation efficiency of approximately 2.5 ${\times}$ $10^3$ ${\sim}$ 5.5 ${\times}$ $10^4$ CFU/${\mu}g$ DNA was obtained where conditions of 0.5 M sucrose electroporation buffer, 1.8 kV pulse voltage and 5 ms time constant were applied. The common conditions developed in this study will make transformation of various Lactobacillus spp. easier than previous procedures.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1399-1403
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• 1998

We isolated wild lactic acid bacteria from kimchi and identified as Lactobacillus brevis by using API 50 CHL Kit, some morphological and physiological tests. In order to evaluate the acid tolerance of Lactobacillus brevis, its survivals rate, glycolysis assay, membrane permeability, and pH profiles of $H^+-ATPase$ were also determined. When Lactobacillus brevis were incubated in Lactobacilli MRS broth adjusted to various levels of pH for 2 hours, the decreases in its population at pH 4.0 and 3.0 were about 2.61 log cycles/mL and 5.89 log cycles/mL, respectively, but there was no decrease at pH 6.0 and 5.0. Glycolysis by Lactobacillus brevis had optimal pH about 6.5 and glucose degradation was reduced by 50% at a pH of 5.2. $Mg^{++}$ release from Lactobacillus brevis cells in medium with pHs of 4.0 and 3.0 was 24.3 and 71.2% of totals, respectively. The $H^+-ATPase$ of Lactobacillus brevis showed a maximal activity between pH values of approximately 6.5 to 7.0.

• Microbiology and Biotechnology Letters
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• v.22 no.5
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• pp.544-549
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• 1994

Lactobacilli proliferating in fermented sausages of the specific ripening conditions were isolated from fermented sausages, manufactured in the absence of an added starter, during ripening under controlled temperature-humidity conditions. Based on morphological, physiological and bio- chemical characteristics and carbohydrate fermentation of isolated strains, three strains of isolates were identified as Lactobacillus curvatus, two strains as Lactobacillus sake. Optimal temperature and pH for growth of isolated strains were 30$\circ$C and pH 6.0~7.0, respectively. These strains were salt tolerant, multiplying in the presense of 6~8% NaCl.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.3
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• pp.456-464
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• 2020

Objective: The aim of this study is to investigate the effect of oral spray with probiotics on the intestinal development and microflora colonization of hatched ducklings. Methods: In Exp. 1, an one-way factorial design was used to study the antibacterial activity of the probiotics and metabolites on Escherichia coli (E. coli) without antimicrobial resistance. There were four experimental groups including saline as control and Lactobacillus, Bacillus subtilis, combined Lactobacillus and Bacillus subtilis groups. In Exp. 2, 64-day-old ducklings were allotted to 2 treatments with 4 replicated pens. Birds in the control group were fed a basal diet supplemented with Lactobacillus fermentation in the feed whereas birds in the oral spray group were fed the basal diet and administrated Lactobacillus fermentation by oral spray way during the first week. Results: In Exp. 1, the antibacterial activities of probiotics and metabolites on E. coli were determined by the diameter of inhibition zone in order: Lactobacillus>combined Lactobacillus and Bacillus subtilis>Bacillus subtilis. Additionally, compared to E. coli without resistance, E. coli with resistance showed a smaller diameter of inhibition zones. In Exp. 2, compared to control feeding group, oral spray group increased (p<0.05) the final body weight at d 21 and average daily gain for d 1-21 and the absolute weight of the jejunum, ileum and total intestine tract as well as cecum Lactobacillus amount at d 21. Conclusion: Lactobacillus exhibited a lower antibacterial activity on E. coli with resistance than E. coli without resistance. Oral spray with Lactobacillus fermentation during the first week of could improve the intestinal development, morphological structure, and microbial balance to promote growth performance of ducklings from hatch to 21 d of age.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.316-322
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• 2011

Lactobacillus sakei OPK2-59 isolated from kimchi was found to have ${\gamma}$-aminobutyric acid (GABA) producing ability and glutamate decarboxylase (GAD) activity. When the Lactobacillus sakei OPK2-59 was cultured in MRS broth with 59.13 mM and 177.40 mM monosodium glutamate (MSG), the optimum temperature range and pH for growth were $25-37^{\circ}C$ and pH 6.5, respectively. GABA conversion rates in MRS broth with 59.13 mM and 177.40 mM MSG were 99.58% and 31.00%, respectively at $25^{\circ}C$ and 48 h of cultivation. By using the cell free extract of Lactobacillus sakei OPK2-59, MSG was converted to GABA and the conversion rate was 78.51% at $30^{\circ}C$, pH 5. Conversion of MSG to GABA was enhanced by adding salts such as $CaCl_2$, $FeCl_3$, $MgCl_2$. These data suggest that the ability of Lactobacillus sakei OPK2-59 to produce GABA results from the activity of GAD in the cells and GABA conversion by the cell extract containing GAD can be enhanced by $CaCl_2$, $FeCl_3$, $MgCl_2$.

• Korean journal of food and cookery science
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• v.1 no.1
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• pp.18-26
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• 1985

The effect of 0.05% Sorbic acid on the Kimchi fermentation and stability of ascorbic acid were investigated at 23~$25^{\circ}C$. The results are as follows. 1. Kimchi with sorbic acid showed a higher pH and a lower total acidity in general, as compared with the control Kimchi. 2. Kimchi with sorbic acid contains more ascorbic acid for the fermentation period. Especially showed the higher hydro ascorbic acid than control Kimchi. 3. The Lacticacid bacteria isolated from Baechu Kimchi and Dongchimi are identifi-ed as Leuconostoc mesenteroides, Lactobacillus Plantarum, Lactobacillus brevis, Streptococcus faecalis, Pediococcus pentosaceus. 4. The effect of sorbic acid upon the growth of Lactic acid bacteria and acid prod-ucibility is found least in Lactobacillus Plantarum, and most in Leuconostoc mesenter-oides. 5. The changes of Lacticacid bacteria occured during Kimchi fermentation curbed Leuconostoc mesenteroides most of all in Baechu Kimchi and Dongchimi with sorbic acid, while the least influence was had on Lactobacillus plantarum. Expecially, yeast wae completely curbed. 6. A result of a sensory evaluation reveals that a better taste is derived from the control Kimchi, with a significant difference, in pH4~4.3, whereas from Kimchi with sorbic acia in pH 3.7~3.8.