• Title/Summary/Keyword: MRS broth

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Characterization of Bacteriocin Produced by Enterococcus faecium MJ-14 Isolated from Meju

  • Lim, Sung-Mee;Park, Mi-Yeon;Chang, Dong-Suck
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.49-57
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    • 2005
  • Enterococcus faecium MJ-14, having strong antilisterial activity, was isolated from Korean fermented food, Meju. MJ-14 showed the same phenotypic characteristics, but different sugar utilization, as reference strain, E. faecium KCCM12118. It could utilize D-xylose, amygdaline, and gluconate, whereas E. faecium KCCM12118 could not. Optimal condition for bacteriocin production by E. faecium MJ-14 was at $37^{\circ}C$ and pH 7.0. Bacteriocin activity appeared in mid exponential phase and increased rapidly up to stationary phase. Activity was significantly promoted in MRS broth containing 3.0% glucose, 1.5% lactose, 2.0% peptone, or 1.5% tryptone. Bacteriocins effectively inhibited Enterococcus faecalis and Listeria spp. of Gram-positive bacteria, and Helicobacter pylori of Gram-negative bacteria, but did not inhibit yeasts and molds. They were stable against heat (for 30 min at $100^{\circ}C$), pH (3.0-9.0), long-term storage (for 60 days at 4 or $-20^{\circ}C$), and enzymatic digestion by catalase, proteinase K, papain, lysozyme, trypsin, chymotrypsin, and lipase, etc. Bacteriocin activity was completely inhibited by protease and pepsin, and 50% by ${\alpha}$-amylase. Studies on PCR detection of enterocin structural genes revealed bacteriocins are identical to enterocins A and B.

Isolation and Identification of Lactobacillus plantarum CIB 001 with Bile Salt Deconjugation Activity from Kimchi (김치로부터 담즙산 분해능이 우수한 Lactobacillus plantarum CIB 001의 분리 및 동정)

  • Cha, Sang-Do;Kim, Tae-Woon;Lee, Dong-Hee
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.222-226
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    • 2010
  • This study was carried out to isolate and characterize the Lactobacillus plantarum with bile salt deconjugation activity that was isolated from Kimchi. Some isolates were selected and identified as L. plantarum by 16S rRNA gene sequence and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of whole cell protein patterns. They were assayed to determine their capacities to express bile salt hydrolase (BSH) activity. Among the identified strains, L. plantarum CIB 001 showed the highest level of BSH activity. Then, resistance to gastric acidity and bile condition were analyzed for further characterization. This strain was able to maintain viability for 1h at pH 2.0 and to survive in a MRS (deMan, Rogosa, and Sharpe) broth with 1.0% of bile acids. L. plantarum CIB 001 would potentially be useful in the food industry as probiotics.

In vitro selection of lactic acid bacteria for probiotic use in pig (양돈용 생균제 균주개발을 위한 유산균주 선발)

  • Ryu, Ji-Sook;Han, Sun-Kyung;Shin, Myeong-Soo;Lee, Wan-Kyu
    • Korean Journal of Veterinary Service
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    • v.32 no.1
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    • pp.33-41
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    • 2009
  • In order to develop probiotic strain for pigs, Lactobacillus spp. (527 isolates), Streptococcus spp. (95 isolates) and Bifidobacterium spp. (25 isolates) were isolated from the feces of 35 pigs. These isolates were tested through in vitro experiment such as acid tolerance at pH 2.0 (Lactobacillus spp. and Streptococcus spp.) or pH 3.0 (Bifidobacterium spp.), bile tolerance in MRS broth containing 0.3% (w/v) Oxgall, heat resistance at $70^{\circ}C$ and $80^{\circ}C$ for 5 min, antibiotic resistance, antimicrobial activity against pathogenic bacteria and Caco-2 cell adherence assay. Finally ten most superior strain (5 Lactobacillus spp. strain, 3 Bifidobacterium spp. strain and 2 Streptococcus spp. strain) were selected as potential candidate for probiotic use in pig industry. It could be used as an alternative to antibiotics in feed additives.

Isolation of a Lactococcus lactis Strain Producing Anti-staphylococcal Bacteriocin

  • Yang, Jung-Mo;Moon, Gi-Seong
    • Food Science of Animal Resources
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    • v.38 no.6
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    • pp.1315-1321
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    • 2018
  • Bacteriocin is ribosomally synthesized by bacteria and inhibits closely related species. In this study we aimed at isolating lactic acid bacteria producing bacteriocin presenting anti-staphylococcal activity. A Lactococcus lactis strain was isolated from kimchi for the purpose and identified by 16S rRNA gene sequencing. As preliminary tests, optimal culture conditions, stabilities against heat, solvents, and enzymes treatments, and type of action (bacteriostatic or bactericidal) of the bacteriocin were investigated. The optimal culture conditions for production of the bacteriocin were MRS broth medium and $25^{\circ}C$ and $30^{\circ}C$ culture temperatures. The bacteriocin was acidic and the activity was abolished by a protease treatment. Its stability was maintained at $100^{\circ}C$ for 15 min and under treatments of various organic solvents such as methanol, ethanol, acetone, acetonitrile, and chloroform. Finally, the bacteriocin showed bactericidal action against Staphylococcus aureus where 200 AU/mL of the bacteriocin decreased the viable cell count (CFU/mL) of S. aureus by 2.5 log scale, compared with a control (no bacteriocin added) after 4-h incubation.

Characterization of the Recombinant Glutamate Decarboxylase of Lactobacillus brevis G144 Isolated from Galchi Jeotgal, a Korean Salted and Fermented Seafood

  • Kim, Jeong A;Park, Ji Yeong;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.9-17
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    • 2021
  • A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5'-phophate. The Km and Vmax of GAD were 8.6 mM and 0.01 mM/min, respectively.

Lactic Acid Bacteria from Gamecock and Goat Originating from Phitsanulok, Thailand: Isolation, Identification, Technological Properties and Probiotic Potential

  • Hwanhlem, Noraphat;Salaipeth, Lakha;Charoensook, Rangsun;Kanjan, Pochanart;Maneerat, Suppasil
    • Journal of Microbiology and Biotechnology
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    • v.32 no.3
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    • pp.355-364
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    • 2022
  • From independent swab samples of the cloaca of indigenous gamecocks (CIG), anus of healthy baby goats (AHG), and vagina of goats (VG) originating from Phitsanulok, Thailand, a total of 263 isolates of lactic acid bacteria (LAB) were collected. Only three isolates, designated C707, G502, and V202, isolated from CIG, AHG, and VG, respectively, exhibited an excellent inhibitory zone diameter against foodborne pathogenic bacteria when evaluated by agar spot test. Isolates C707 and G502 were identified as Enterococcus faecium, whereas V202 was identified as Pediococcus acidilactici, based on 16S rRNA sequence analysis. When foodborne pathogenic bacteria were co-cultured with chosen LAB in mixed BHI-MRS broth at 39℃, their growth was suppressed. These LAB were found to be capable of surviving in simulated stomach conditions. Only the isolate G502 was able to survive in the conditions of simulated intestinal juice. This research suggests that selected LAB could be used as a food/feed supplement to reduce foodborne pathogenic bacteria and improve the safety of animal-based food or feed.

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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GABA Productivity in Yoghurt Fermented by Freeze Dried Culture Preparations of Lactobacillus acidophilus RMK567 (Lactobacillus acidophilus RMK567의 동결건조 컬쳐로 제조한 요구르트에서 GABA 생성력)

  • Lim, Sang-Dong;Yoo, Sung-Ho;Yang, Hae-Dong;Kim, Sang-Ki;Park, Seung-Yong
    • Food Science of Animal Resources
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    • v.29 no.4
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    • pp.437-444
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    • 2009
  • ${\gamma}-Aminobutyric$ acid (GABA) producing lactic acid bacteria, Lactobacillus acidophilus RMK567 was cultivated in 50 L of sterilized MRS broth using a fermenter at $40^{\circ}C$ for 24 h. The cell number was increased to $10.04{\pm}0.13$ Log CFU/mL with a growth rate constant (k) of 0.454 generation/h and a generation time (g) of 2.303 h after a lapse of a lag phase (L) of 5.16 h. A total of 487 g of cell paste with 40.5% moisture was harvested with viable cell number of 12.48 Log CFU/g cell paste. The cell pastes after preparation with glycerol, glucose, and polydextrose as cryo-protectants were lyophilized under a vacuum of 84 m torr. A total of 408 g of freeze dried (FD) cell powders were mixed with a commercial strain of Streptococcus thermophilus to prepare of three types FD starter cultures with the viable cell numbers of 12.42 (FDA-GY), 12.60 (FDBGG) and 12.91 (FDC-GP) Log CFU/g. During preservation the FD cultures at -$18^{\circ}C$, the cell viability of the FD starter cultures were rapidly dropped to below 3.24% of the day of storage. No significant difference was found in the cell viabilities among three types of FD starters cultures, but significant difference (p<0.01) was found in storage periods. Yoghurts fermented through FD starter culture of L. acidophilus RMK567 were determined to contain $155.16{\pm}8.53$ ppm, $243.82{\pm}4.27$ ppm, and $198.64{\pm}23.46$ ppm of GABA, respectively. This study shows that GABA production activity of L. acidophilus RMK567 is not affected during the freeze drying process and would be available for commercial production of yoghurt containing high GABA content.

Studies on the Utilization of Wastes from Fish Processing I - Characteristics of Lactic Acid Bacteria for Preparing Skipjack Tuna Viscera Silage (수산물 가공부산물의 이용에 관한 연구 I -가다랭이 내장 발효 silage 제조를 위한 유산균주의 배양특성)

  • YOON Ho-Dong;LEE Doo-Seog;JI Cheong-Il;SUH Sang-Bok
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.1
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    • pp.1-7
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    • 1997
  • In order to utilize fish by-products from the skipjack tuna (Katsuwonus pelamis) canning manufactures Lactobacillus buigaricus KCTC 3188 and L. piantarum KCTC 1048 were used as a starter culture for the preparation of fermented fish silage with skipjark tuna viscera. The optimum temperature and pH on barterial growth and lactic acid production of L. bulgaricus and L. plantarum in MRS broth were $35^{\circ}C$ and around pH 6.0, respectively. And the optimum concentrations of the carbohydrate sources added to the broths were $7\%$ for dextrose and $10\%$ for molasses on the basis of total weights of skipjack tuna viscera. The pH of acid treated skipjack tuna viscera silage (ASS) slightly increased from 4.0 to 4.5, while that of fermented skipjack tuna viscera silages by the use of lactic acid bacterias (FSS) was significantly declined from 5.9 to about 40 after 42 days of storage at $35^{\circ}C$. Though the content of volatile basie nitrogen (VBN) in ASS was lower than those of FSS after 42 days of storage at $35^{\circ}C$, VBN content in silages slightly increased from an initial value of $62\~65{\cdot}mg/100g$ to final value of $113\~155\;mg/100g$ over 42 days. The fermented silage by L. piantarum reached a maximum concentration of amino nitrogen and showed $81\%$ of hydrolysis degree after 4 days of storage at $35^{\circ}C$.

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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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