• Title/Summary/Keyword: $^1H$-MRS

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Changes in Acidity and Distributions of the Vancomycin-Resistant Lactic Acid Bacteria in the Kimchi Fermented at Different Temperatures (발효 온도에 따른 김치의 산도 변화와 Vancomycin 내성 젖산균의 분포)

  • 정의숙;김기환;신원철;송광영;윤성식
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.249-255
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    • 2004
  • Chinese cabbage ('Baechu') Kimchi was fermented at the three different temperatures right after it was prepared. Samples were taken everyday for measuring bacterial populations, pH, and titratable acidity through the whole periods of fermentation up to 50 days. pH values and developed acidity were significantly affected by the fermenting temperatures of 4, 10, and $20^{\circ}C$, suggesting that different bacterial flora has been established by the temperatures exposed. The modified MRS agar containing vancomycin (300 $\mu$g/mL) was used for isolating the vancomycin-resistant LAB strains and 127 isolates were finally obtained. Of the LAB isolates, 13 isolates were subjected to the identification experiments based on the biochemical characteristics and the molecular-typing approach, an ITS-PCR, whether they belong to the genus Leuconostoc or not. The data obtained from API 50 CHL kit resulted that six isolates were identified as the members of Leuconostoc and six as Lactobacillus brevis strains except for a single isolate YKI 30-0401, which was not able to be identified because its biochemical traits were not matched to the database of API 50 CHL kit. It was noted that some isolates were distinct in a couple of some biochemical characteristics compared with those of the reference Leuconostoc species. To overcome the limitations experienced in the commercial identification products above, an ITS-PCR experiment was also conducted for the isolates, resulting that eight isolates belong to Leu. mesenteroides ssp. mesenteroides or dextranicum with a single band of 564 bp, and four to L. brevis strains. The ITS-PCR profiles clearly differentiated the closely-related LAB isolates for which same results were obtained by the biochemical method. This molecular approach, however, failed to produce the amplicons for the YKI 20-1003, leaving the strain unidentified. Judging from the identification data obtained in the Kimchi fermented at $4^{\circ}C$ or $10^{\circ}C$, Leuconostoc spp. including Leu. mesenteroides/dextranicum were likely predominant species in the earlier stage and L. brevis occurred at the high level through the whole period. By contrast, L. brevis, as one of the major flora, possibly lead the fermentation from the beginning in the Kimchi fermented at $20^{\circ}C$.}C$.

Effect of the amino acid mixture on freeze-drying and preservation of Lactobacillns casei YIT 9018 (아미노산 혼합용액이 Lactobacillus casei YIT 9018의 동결건조 및 저장성에 미치는 영향)

  • 윤성식;이해옥;유주현
    • Microbiology and Biotechnology Letters
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    • v.14 no.5
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    • pp.421-426
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    • 1986
  • This study was attempted to find out effective storage methods of Lactobacillus casei YIT 9018, industrial strain for fermented mil k production, without severe bacterial death and activity deteriorations. The cryoprotective effect of the ammo acid mixture consisting of glycine and DL-g1utamic acid on the test strain were examined and also compared with those other protectants already reported. The apparent protective effect by the amino acid mixture was observed to controls. Both glycine and DL-glutamic acid prevented the freezing death of test strain and his effect of 1. casei YIT 9018 had reached stationary stage in MRS-broth 18h after inoculation. Cells harvested from stationary stage were most resistant to freezing damage. The viability of the test strain was affected by rehydration media and the recovery of viable cells was increased about threefold when amino acid mixture was used for rehydration. The presence of non-fat milk solid (NFMS), sucrose and lactose in amino acid mixture increased viability of the test strain up to 85%. In this case, optimal concentrations of NFMS, sucrose and lactose were 10%, 7.5-10%, 7.5-10%, respectively.

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Depletion of Nitrite by Lactic Acid Bacteria Isolated from Kimchi(II) (김치에서 분리한 유산균에 의한 아질산염 소모(II))

  • 오창경;오명철;현재석;최우정;이신호;김수현
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.4
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    • pp.556-562
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    • 1997
  • Depletion of nitrite by lactic acid bacteria isolated from Kimchi and the effects of pH and temperature on depletion of nitrite in Lactobacillus MRS broth were investigated. Depletion of nitrite by Lactobacillus plantarum and Lactobacillus sake was low in initial stage of growth but increased during growth at 15 and 2$0^{\circ}C$. L. plantarum and L. sake depleted than 90 and 75% of nitrite(250$\mu\textrm{g}$/$m\ell$) in 1 day of growth at 25 and 3$0^{\circ}C$, respectively, but depleted almost all of nitrite in 2 days. While the effect of Leuconostoc mesenteroides (150$\mu\textrm{g}$/$m\ell$) was lower compared to Lactobacillus(250$\mu\textrm{g}$/$m\ell$). In addition, even high concentrations(600 and 900 $\mu\textrm{g}$/$m\ell$) of nitrite was depleted at 2 days of growth by L. plantarum. pH of growing broth decreased as the increase of growth time and temperature, and depletion of nitrite in- creased as the decrease of pH of growing culture. However, pH of broth related with the decrease of depletion of nitrite at 25 and 3$0^{\circ}C$, but not at 15 and 2$0^{\circ}C$. Therefore, depletion of nitrite was greatly affected by growth temperature rather than by pH.

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Development of Cabbage Juice Medium for Industrial Production of Leuconostoc mesenteroides Starter

  • Jeong, Eun Ji;Moon, Dae Won;Oh, Joon Suk;Moon, Jin Seok;Seong, Hyunbin;Kim, Kwang Yup;Han, Nam Soo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2112-2118
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    • 2017
  • Leuconostoc mesenteroides is used as a starter to produce high-quality kimchi products. In this study, an efficient and economical cabbage juice medium (CJM) was developed by process optimization of cabbage extraction and pasteurization and by compositional supplementation of various lacking nutrients. The pasteurized cabbage juice was determined to be a good medium candidate to cultivate L. mesenteroides, showing maximal cell numbers ($9.85{\times}10^8CFU/ml$) after 24 h. Addition of sucrose and yeast extract with soy peptone resulted in increment of bacterial cell counts in CJM, showing the supplementing effect of the lacking nutrients. Furthermore, addition of shell powder gave a protective effect on bacterial cells by preventing pH decline and organic acid accumulation in CJM, resulting in a 2-fold increase of bacterial counts. The optimized composition of CJM was 70% cabbage juice diluted with water, 0.5% (w/v) sucrose, 1% (w/v) yeast extract, 1% (w/v) soy peptone, and 1.5% (w/v) ark shell powder. The CJM developed in this study was able to yield a comparable level of bacterial counts with MRS medium and reduced the cost by almost 10-fold.

Strain Improvement of Leuconostoc paramesenteroides as a Acid-Resistant Mutant and Effect on Kimchi Fermentation as a Starter. (Leuconostoc paramesenteroides의 내산성 변이주로의 개량과 starter로의 첨가효과)

  • 김영찬;정은영;김은해;정대현;이옥숙;권태종;강상모
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.151-160
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    • 1998
  • The Leuconostoc paramesenteroides dominated at refrigeration temperature range was isolated from kimchi, and improved its growth properties by mutation for competitive growth against Lactobacillus plantarum at lower pH. It was found that the minimal pH for the wild type Leuconostoc paramesenteroides Pw growth was pH 4.5 adjusted with HCI and pH 5.0 adjusted with organic-mixture (lactic acid:acetic acid=1:2), respectively. The mutant P-100 could grow in pH 4.0, 4.5, respectively, in MRS broth. Two strains Pw and P-100 were added into kimchi as starter and compared the quality characteristics of kimchi. The total acceptability of Pw and P-100 inoculated kimchi were evaluated better than that of control kimchi (no starter added) by sensory test and extended the optimal pH range of kimchi up to about 2.2, 2.5 times, respectively. In kimchi added P-100, the succinic acid was more abundant than others and the total number of Lactobacillus plantarum was down about 2.5 times in contrast to control kimchi.

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H2O2 Generating Ability and Multi-Drug Resistance of Lactic Acid Bacteria Required for Long-Term Inpatient Treatment with Antibiotic Resistance

  • Yuk, Young Sam
    • International journal of advanced smart convergence
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    • v.11 no.4
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    • pp.227-239
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    • 2022
  • Purpose: In our study, in order to find lactic acid bacteria (LAB) with multi-drug resistance to antibiotics, we isolated 140 strains from 15 types of kimchi commercially available in Korea and 20 types of Kimchi made at home from January to December in 2016, and investigated their H2O2 generating ability and multi-drug resistance to antibiotics. Methods: In order to observe the H2O2 generation ability of LAB, we performed the experiment with methods such as Rabe, Hillier, and Kang. To test the antibacterial susceptibility of LAB, we used the disc agar diffusion method using MRS agar (Difco, USA) according to the CLSI and WHO test methods. There are 18 types of antibiotic discs used. Results: Out of the total numbers of 140 strains, 6 strains of Ent. Faecium, 25 strains of L. plantarum, 1 strain of L. rhamnosus, 3 strains of L. sakei, 1 strain of L. acidophilus, 1 strains St. thermophilus, and 7 of unidentified strains generated H2O2. The antibiotic susceptibility of Ent. Faecium indicated SXT, OX, NA, and E; and the antibiotic susceptibility of L. plantarum indicated NA; and the antibiotic susceptibility of St. thermophilus indicated NA, CC, RA, CTT, CM, and P ; and the antibiotic susceptibility of L. rhamnosus indicated SXT, VA, NA and CTT; and the antibiotic susceptibility of 6 strains of L. sakei indicated SXT, OX, NOR, NA, CTT and CIP, all indicating antibiotic resistance. In the case of multi-drug resistance to antibiotics for 53 strains of L. antarum, 8-drug resistance was the most common with 25 strains, followed by 7-drug-resistant strains with 18 strains, 9-drug-resistant strains with 4 strains, 6-drug-resistant strains with 3 strains, 5-drug-resistant strains with 2 strains, and 17-drug-resistant strains with 1 strain. In the case of multi-drug resistance to antibiotics for Ent. Faecium 27 strains, 9-drug resistance was most commonly identified as 9 strains, 8-drug resistance was identified as 6 strains, 7- and 11 drug resistances were identified as 4 strains each, and 4- and 6-drug resistances were identified as 1 strain each. Conclusion: Ent. Faecium, L. plantarum, L. rhamnosus, L. sakei, and St. thermophilus, shown to have anantibacterial activity in previous studies on LAB and shown to have and H2O2 generating ability, antibiotic resistance and multi-drug resistance in this study, are expected to be able to play an excellent role for long-term inpatients to use as an alternative to antibiotics and to cope with emerging antibiotic resistance.

Evaluations of Spectral Analysis of in vitro 2D-COSY and 2D-NOESY on Human Brain Metabolites (인체 뇌 대사물질에서의 In vitro 2D-COSY와 2D-NOESY 스펙트럼 분석 평가)

  • Choe, Bo-Young;Woo, Dong-Cheol;Kim, Sang-Young;Choi, Chi-Bong;Lee, Sung-Im;Kim, Eun-Hee;Hong, Kwan-Soo;Jeon, Young-Ho;Cheong, Chae-Joon;Kim, Sang-Soo;Lim, Hyang-Sook
    • Investigative Magnetic Resonance Imaging
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    • v.12 no.1
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    • pp.8-19
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    • 2008
  • Purpose : To investigate the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar nuclear Overhauser effect/enhancement (NOE) interaction through 2D- correlation spectroscopy (COSY) and 2D- NOE spectroscopy (NOESY) techniques. Materials and Methods : All 2D experiments were performed on Bruker Avance 500 (11.8 T) with the zshield gradient triple resonance cryoprobe at 298 K. Human brain metabolites were prepared with 10% $D_2O$. Two-dimensional spectra with 2048 data points contains 320 free induction decay (FID) averaging. Repetition delay was 2 sec. The Top Spin 2.0 software was used for post-processing. Total 7 metabolites such as N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), lutamine (Gln), glutamate (Glu), myo-inositol (Ins), and lactate (Lac) were included for major target metabolites. Results : Symmetrical 2D-COSY and 2D-NOESY pectra were successfully acquired: COSY cross peaks were observed in the only 1.0-4.5 ppm, however, NOESY cross peaks were observed in the 1.0-4.5 ppm and 7.9 ppm. From the result of the 2-D COSY data, cross peaks between the methyl protons ($CH_3$(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methylene protons (CH2(3,$H{\alpha}$)) at 2.50ppm and methylene protons ($CH_2$,(3,$H_B$)) at 2.70 ppm were observed in NAA. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. From the result of 2-D NOESY data, cross peaks between the NH proton at 8.00 ppm and methyl protons ($CH_3$) were observed in NAA. Cross peaks between the methyl protons ($CH_3$(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methyl protons (CH3) at 3.03 ppm and methylene protons (CH2) at 3.93 ppm were observed in Cr. Cross peaks between the methylene protons ($CH_2$(3)) at 2.11 ppm and methylene protons ($CH_2$(4)) at 2.35 ppm, and between the methylene protons($CH_2$ (3)) at 2.11 ppm and methine proton (CH(2)) at 3.76 ppm were observed in Glu. Cross peaks between the methylene protons (CH2 (3)) at 2.14 ppm and methine proton (CH(2)) at 3.79 ppm were observed in Gln. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. Conclusion : The present study demonstrated that in vitro 2D-COSY and NOESY represented the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar NOE interaction. This study could aid in better understanding the interactions between human brain metabolites in vivo 2DCOSY study.

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Screening and Characterization of Probiotic Strains for Prevention of Bacterial Fish Diseases (어류의 세균성 질병 예방을 위한 Probiotic균주의 선발 및 특성)

  • 허문수;양병규;전유진
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.129-134
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    • 2003
  • The purpose of the present study was to screen the effective of lactic acid bacteria (LAB) as probiotics, which are able to protect aquacultural fish pathogenic bacteria, and investigate their characterization. Twenty strains of lactic acid bacteria were isolated from fish intestine, fermented fish foods and kimchis. These bacteria were screened for antagonistic activity against fish pathogenic bacteria. Seven tested LAB strains were able to inhibit the fish pathogenic bacteria, including Vibrio anguillarum, Edwardsiella tarda, and Streptococcus sp.. Of the probiotic candidates, BK19 strain isolated from fermented pollack viscera indicated the largest inhibition activity. Moreover, this strain showed a resistance over low pH and antibiotic agents. Therefore this probiotic candidate BK19 was finally selected and identified as a probiotic strain. This particular probiotic bacteria was identified as Lactobacillus sakei BK19 by biochemical characteristics and 165 rRNA PCR amplification.

Inhibition of Lactic Acid Bacteria in Kimchi Fermentation by Nisin

  • CHOI, MIN HO;YUN HEE PARK
    • Journal of Microbiology and Biotechnology
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    • v.8 no.5
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    • pp.547-551
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    • 1998
  • Sixty isolates of lactic acid bacteria found in kimchi, a traditional Korean dish of fermented vegetables, were tested for nisin sensitivity. Of the sixty isolates, all belonging to the genera Leuconostoc, Lactobacillus, and Pediococcus, fifty isolates were sensitive to nisin at a concentration of 100 IU/$m\ell$, and four isolates appeared to be resistant to nisin. This demonstrated that the nisin sensitivity of lactic acid bacteria found in kimchi varied considerably among isolates. In MRS broth containing nisin at concentrations of 100 to 300 IV/$m\ell$, the growth of sensitive isolates of Leuconostoc mesenteroides and Lactobacillus plantarum was inhibited for two to three days at 2$0^{\circ}C$. When nisin was added to kimchi preparations at a concentration of 100 IU/$m\ell$, the growth of lactic acid bacteria was delayed and reached a maximum two days later than that in kimchi without nisin. These results suggest the possible use of nisin in kimchi preparation, at recommended levels, to control the lactic acid fermentation. Scanning electron micrographs of a sensitive isolate L. plantarum revealed the formation of pores on cell surfaces followed by rapid cell wall destruction 1 h after the addition of nisin.

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Enhanced Production of ${\gamma}$-Aminobutyric Acid Using Rice Bran Extracts by Lactobacillus sakei B2-16

  • Kook, Moo-Chang;Seo, Myung-Ji;Cheigh, Chan-Ick;Pyun, Yu-Ryang;Cho, Seok-Cheol;Park, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.763-766
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    • 2010
  • An efficient and simple fermentation process was developed for the production of ${\gamma}$-aminobutyric acid (GABA) by Lactobacillus sakei B2-16. When the L. sakei B2-16 was cultivated in the rice bran extracts medium containing 4% sucrose, 1% yeast extract, and 12% monosodium glutamate, the maximum GABA concentration reached 660.0 mM with 100% conversion yield, showing the 2.4- fold higher GABA concentration compared with the modified MRS medium without the rice bran extracts. The GABA production was scaled-up from a laboratory scale (5 l) to a pilot (300 l) and a plant (5,000 l) scale to investigate the application possibility of GABA production to industrial fields. The production yields at the pilot and plant scales were similar to the laboratory scale using rice bran extracts medium, which could be effective for the low-cost production of GABA.