• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1216-1222
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• 2017

To develop starters for the production of functional foods or materials, lactic acid bacteria producing ${\gamma}-aminobutyric$ acid (GABA) were screened from jeotgals, Korean fermented seafoods. One isolate producing a high amount of GABA from monosodium $\text\tiny{L}$-glutamate (MSG) was identified as Enterococcus avium by 16S rRNA gene sequencing. E. avium M5 produced $18.47{\pm}1.26mg/ml$ GABA when incubated for 48 h at $37^{\circ}C$ in MRS broth with MSG (3% (w/v)). A gadB gene encoding glutamate decarboxylase (GAD) was cloned and overexpressed in E. coli BL21 (DE3) using the pET26b (+) expression vector. Recombinant GAD was purified through a Ni-NTA column and the size was estimated to be 53 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 4.5 and $55^{\circ}C$and the activity was dependent on pyridoxal 5'-phosphate. The $K_m$ and $V_{max}$ values of GAD were $3.26{\pm}0.21mM$ and $0.0120{\pm}0.0001mM/min$, respectively, when MSG was used as a substrate. Enterococcus avium M5 secretes a lot of GABA when grown on MRS with MSG, and the strain is useful for the production of fermented foods containing a high amount of GABA.

• Microbiology and Biotechnology Letters
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• v.28 no.5
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• pp.279-284
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• 2000

This study was per-formed to screen lactic acid bacteria poultry for the probiotic use. Among the previously obtained acid tolerant, 139 strains, 111 strains were selected with MRS medium containing 0.3% oxgall. 34 strains of 111 was re-selected by Gram-staining and acid producing ability. These strains was identified by MIDI Sherlock Microbial Identification System. Among the identified 34 strains Lactobacillus fermenum YL-3 was selected for the final pro-biotic use because of the good growth and high survival rate at pH 2.0. 60%, 50% and 40% cells of Lactobacillus fermentum YL-3 survived at pH 3.0, 2.5 and 2.0, respectively. More than $10^{7}$ / CFU/ml survived when exposed with the number of $10^{8}$ CFU/ml at pH 2.0 after 12 hr. L.fermenum YL-3 maintained growth in MRS broth containing 0.3, 0.5, 1.0 and 2.0% oxgall for 24 hr. L.fermenum YL-3 showed an inhibitory effect against pathogenic strains of Sal. enteritidis and E. coli O157:H7. In mixed culture with L.fermenum YL-3 Sal. enteritidis lost ability com-pletely in 15 hrs and E. coil O157:H7 in 16 hrs.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.4
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• pp.549-555
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• 1997

Twenty species of lactic acid bacteria were isolated from Kimchi. Six species were identified as Lactobacillus sake, ten species as Leuconostoc mesenteroides and four species as Lactobacillus plantarum. The ability of these isolates to deplete nitrite during growth at 15, 20, 25 and 3$0^{\circ}C$ in MRS broth containing 250$\mu\textrm{g}$/$m\ell$ of nitrite was examined. Depletion of nitrite was high as the order of L. plantarum, L. sake and L. mesenteroides at all temperature tested, and the depletion activity was increased as the increase of growth temperatures. Especially, almost all of nitrites were utilized by L. plantarum during growth at all temperature ranges tested. L. plantarum and L. sake required induction periods for adapting to nitrite, but L. plantarum was remarkably depleted nitrite after two days of growth at 15$^{\circ}C$ and one day at 2$0^{\circ}C$, L. sake after one day of growth at both temperatures. Whereas, L. mesentero-ides did not require those periods at all temperature ranges tested.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.420-426
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• 2004

In this study, among 10 medicinal plants extracted with 50%-ethanol, antimicrobial activities measured by paper disc method were showed in Scutellaria baicalensis (Sb) on Leu mesenteroides and in Phellodendron amurene (Pa) on Lac. plantarum and Leu. mesenteroides. While 0.05∼0.2% of Pa extracts showed also relatively strong growth inhibition of both strains of lactic acid bacteria cultured in MRS broth for 24 hours at 3$0^{\circ}C$, Sb extract at concentration of 0.1% showed similar inhibitory effect on Leu mesenteroides to that of 0.05% of Pa extract. Addition of 0.02∼0.04% of Pa extracts to kimchi lowered effectively extents of pH decrease and acidity increase, and numbers of lactic acid bacteria and total bacteria compared to those of control during fermentation at 4$^{\circ}C$. This extending effect on fermentation period was obtained by adding 0.04% in case of Sb. Mixed extracts of Pa and Sb (Pa-Sb) by ratio 1 : 1 delayed also fermentation of kimchi at 0.03∼0.04%. In sensory evaluation kimchi containing 0.04% of Pa extract were less sour than kimchi containing 0.02% of Sb extract and/or control at late stage of fermentation.

• Korean Journal of Soil Science and Fertilizer
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• v.48 no.2
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• pp.125-131
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• 2015

Phenyllactic acid (PLA) which is known as antimicrobial compound can be synthesized through the reduction of phenylpyruvic acid (PPA) by lactate dehydrogenase (LDH) of lactic acid bacteria (LAB). LAB producing PLA was isolated from Korea Kimchi and identified to Lactobacillus plantarum SJ21 by 16 rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. plantarum SJ21 was assessed for both the capability to produce the antimicrobial compound PLA and the antifungal activity against four fungal pathogens (Rhizoctonia solani, Aspergillus oryzae, Botrytis cinerea, and Collectotricum aculatum). PLA concentration was investigated to be 3.23mM in CFS when L. plantarum SJ21 was grown in MRS broth containing 5mM PPA for 16 h. PLA production also could be promoted by the supplement of PPA and phenylalanine in MRS broth, but inhibited by the supplement of 4-hydroxyphenylpyruvic acid and tyrosine as precursors. Antifungal activity demonstrated that all fungal pathogens were sensitive to 5% CFS (v/v) of L. plantarum SJ21 with average growth inhibitions ranging from 27.32% to 69.05% (p<0.005), in which R. solani was the most sensitive to 69.05% and followed by B. cinerea, C. aculatum, and A. oryzae. The minimum inhibitory concentration (MIC) for commercial PLA was also investigated to show the same trend in the range from $0.35mg\;mL^{-1}$ (2.11 mM) to $0.7mg\;mL^{-1}$ (4.21 mM) at pH 4.0. The inhibition ability of CFS against the pathogens was not affected by heating or protease treatment. However, pH modification in CFS to 6.5 caused an extreme reduction in their antifungal activity. These results may indicate that antifungal activities in CFS were caused by acidic compounds like PLA or organic acids rather than proteins or peptides molecules.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1798-1807
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• 2002

This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.5
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• pp.834-841
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• 1997

A lactic acid bacteria which showed antimicrobial activity was isolated from dairy products and identified as Lactobacillus sp. according to the morphological, physiological and biochemical properties, which was named Lactobacillus sp. SH 7311. The bacteriocin of Lactobacillus sp. GM 7311 showed a broad range of inhibitory spectrum against some gram positive and negative bacteria. Especially, Proteus mirabilis was highly sensitive to bacteriocin and used as indicator strain for further investigation. The optimal condition for the production of bacteriocin was showed on MRS broth at $37^{\circ}C$ and pM 6.0. Bacteriocin production of this strain cultured under optimal condition was increased late logarithmic phase to early stationary phase. This bacteriocin was fully active at the pH range $2.0\~5.0$, also was stable at $100^{\circ}C$ for 60 min. at pH 5.0, But about $40\%$ of bacteriocin activity was diminished by the treatment of acetone, ethanol, iso-butanol and ethyl ether during 2 hours at $4^{\circ}C$.

• 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.

• Korean Journal of Organic Agriculture
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• v.22 no.3
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• pp.491-502
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• 2014

The present study was conducted to investigate effective starter culture to improve biological activity of Asarum sieboldii. Antibacterial activity, antioxidant activity and reduction of enteric rumen methane production were used as criterions for biological activity. Ground A. sieboldii was added in MRS broth at 10% (w/v) and fermented by different starter cultures. Weissella confusa NJ28, Weissella cibaria NJ33, Lactobacillus curvatus NJ40, Lactobacillus brevis NJ42, Lactobacillus plantarum NJ45 and Lactobacillus sakei NJ48 were used for starter culture strains. Each starter culture was inoculated with 1% (v/v) ratio and fermentation was performed at $30^{\circ}C$ with agitation (150 rpm) for 48 h. MRS broth for the control was employed without starter culture. Then the fermentation growth was dried and extracted using ethyl alcohol. The growth of starter culture was detected at NJ40, NJ42, NJ45 and NJ48. And the highest cell growth was found in NJ40. Antibacterial activity against to Staphylococcus aureus, Listeria monocytogens, Mannheimia haemolytica and Salmonella gallinarum were observed in the extract fermented by NJ40 and NJ45. All treatments showed antioxidant activities, however, there were no significant differences (p>0.05). In in vitro rumen fermentation, negative control (NC) and positive control (PC) were assigned to without extract and with non-fermented A. sieboldii extract. Significant suppression of gas productions were detected in positive control and treatments compared to negative control (p<0.05). However, total volatile fatty acid production was not suppressed. Significant methane reduction per total volatile fatty acid productions were found in positive control and NJ45 treatment (p<0.05). The present study suggested a fermentation of A. sieboldii using NJ45 strain could improve its biological activity and make possible for its use in bio additive for enteric rumen methane mitigation without suppression of animal productivity.

• Food Science of Animal Resources
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• v.20 no.3
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• pp.181-191
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• 2000

젖산균의 생장을 위한 질소, 탄소 공급원으로서 corn steep liquor의 이용 가능성을 시험하고 반응 표면 분석(Response surface metho-dology)을 이용하여 젖산균의 최적 생상 배지 조성을 연구하였다. 반응 표면 분석에서 L.fermentum의 생장배지에 첨가되 corn steep liquor와 yeast extract의 농도(p<0.01) 그리고 corn steep liquor와 yeast extract의 교호작용(P<0.05)이 L.ferm-entum의 생장에 큰 영향을 미치는 것으로 나타났으며, 이때 생균수가 최대인 corn steep liquor의 함량은 10.77%, yeast extract는 3.39%.Tween 80은 1.69%으로 예측되었다. 한편, Lc, lactis ssp. lactis 의 생장배지는 corn steep liquor 의 농도(P<0.01) 그리고 corn steep li-quor와 $\beta$-glycerophosphate disodium salt의 교호작용(P<0.05)인 Lc. lactis ssp. lactis의 생장에 큰 영향을 미치는 것으로 나타났으며 이때 생균수가 최대인 corn steep liquor의 함량은 3.5%, $\beta$-glycerophosphate disodium salt는 4.38%로 예측되었다. MRS broth와 예측된 최적 배지에서 L.fermentum의 젖산과 초산의 생성량은 각각 0.166, 0.114과 0.273, 0.081 M이고 M 17glc broth와 최적배지에서 Lc. lacti ssp. lactis의 젖산과 초산의 생성량은 각각 0.089, 0.003과 0.189, 0.003M이었다. 따라서 corn steep liquor는 L. fermentum와 Lc. lactis ssp, lactis 의 생장을 위해 질소 또는 탄소 공급원으로서 배지에 첨가 될 수 있는 우수한 농업 부산물로 판단되었다.