• Title/Summary/Keyword: MRS broth

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Protease Activity of Lactic Acid Bacteria Isolated from Korean Traditional Fermented Food (전통 발효식품으로부터 Protease 활성을 보유한 유산균의 분리 및 동정)

  • Kook, Moo Chang;Cho, Seok Cheol;Park, Hoon;Kim, Seung Seop;Pyun, Yu Ryang;Choi, Woon Yong;Lee, Hyeon Yong
    • Food Engineering Progress
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    • v.15 no.2
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    • pp.182-187
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    • 2011
  • A proteolytic lactic acid bacterium was isolated from Korean traditional fermented foods. The isolate BV-26, which had a protease activity (24 U/mg-crude protein), was identified as Lactobacillus plantarum by the API 50CHL kit and 16S rDNA analysis (99.9% of homology), and named as L. plantarum BV-26. Cell growth and protease activity of L. plantarum BV-26 was determined in MRS broth using 5L jar fermentor at $30^{\circ}C$. The maximum growth of L. plantarum BV-26 was reached at 18 hr in MRS broth, while protease activity of BV-26 was detectable at 12 hr and the highest activity was obtained after 16 hr cultivation. Therefore, we expect that the proteolytic lactic acid bacteria, L. plantarum BV-26, may be used as a starter for the fermentation of animal feed. Especially, the fermentation of soybean meal with the strain can be applied for improving feed utilization.

Analysis of Antibacterial, Antioxidant, and In Vitro Methane Mitigation Activities of Fermented Scutellaria baicalensis Georgi Extract (발효 황금 뿌리 추출물의 항균, 항산화 효과 및 메탄가스 저감 효과 In Vitro)

  • Marbun, Tabita Dameria;Song, Jaeyong;Lee, Kihwan;Kim, Su Yeon;Kang, Juhui;Lee, Sang Moo;Choi, Young Min;Cho, Sangbuem;Bae, Guiseck;Chang, Moon Baek;Kim, Eun Joong
    • Korean Journal of Organic Agriculture
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    • v.24 no.4
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    • pp.735-746
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    • 2016
  • This study was conducted to investigate the antibacterial, antioxidant, and in vitro greenhouse gas mitigation activities of fermented Scutellaria baicalensis Georgi extract. Seven starter cultures were used, comprising four of lactic acid bacteria and three of Saccharomyces cerevisiae. Ten grams of S. baicalensis Georgi powder was diluted in 90 mL autoclaved MRS broth. Each seed culture was inoculated with 3-10% (v/v) S. baicalensis Georgi MRS broth and incubated at $30^{\circ}C$ for 48 h. Among the starter cultures used, only Lactobacillus plantarum EJ43 could withstand the fermentation conditions. This fermentation broth was dried and extracted with ethanol to assess its antibacterial, antioxidant, and in vitro methane mitigation activities. The extract of S. baicalensis Georgi fermented by L. plantarum EJ43 (SBLp) showed higher antibacterial activity (bigger clear zone) compared to the unfermented S. baicalensis Georgi extract (SB0). SBLp also presented 1.2 folds higher antioxidant activity than SB0. During in vitro rumen fermentation, SBLp showed reduction in methane production compared to SB0 or the control. In conclusion, fermentation by L. plantarum EJ43 may enhance antibacterial and antioxidant activities of S. baicalensis Georgi and decrease enteric methane production.

Characteristics and Purification of Bacteriocin Produced by Lactobacillus acidophilus GP4A (Lactobacillus acidophilus GP4A가 생산하는 박테리오신의 특성 및 정제)

  • Han, Gyeong-Sik;Joo, Gwan-Seok;Kim, Se-Heon
    • Journal of Dairy Science and Biotechnology
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    • v.17 no.1
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    • pp.1-10
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    • 1999
  • A bacteriocin produced by Lab. acidophilus GP4A isolated from fecal contents of pig was characterized. Lab. acidophilus GP4A produced a heat-stable and pH-resistant bacteriocin, which was hydrolyzed by trypsin and pepsin and active against various microorganisms. Lab. acidophilus GP4A produced bacteriocin at maximum rate when grown in MRS broth(pH 6.5${\sim}$7.5) at$37^{\cric}C$ or $40^{\cric}C$. The bacteriocin produced by Lab. acidophilus GP4A inhibited the growth of Lactobacillus delbrueckii subsp. lactis 4794 in early logarithmic phase. The bacteriocin was purified by ammonium sulfate precipitation and Octyl sepharose CL-4B column chromatography. The purification resulted in a final yield of 21.7% and a 13.6-fold increase in the specific activity.

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Probiotic properties and safety assessment of lactic acid bacteria isolated from salt-fermented anchovy (멸치 젓갈로부터 분리된 젖산세균의 프로바이오틱 특성 및 안전성 평가)

  • Lim, Eun-Seo;Kim, Young-Mog;Lee, Eun-Woo
    • Korean Journal of Food Science and Technology
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    • v.48 no.4
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    • pp.306-316
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    • 2016
  • This study was conducted to evaluate the functional characteristics and safety properties of lactic acid bacteria isolated from salt-fermented anchovy, a putative probiotic candidate. The following isolates were identified by biochemical profiles, carbohydrate fermentation patterns, and 16S rRNA sequencing: Enterococcus faecium AJ06, Leuconostoc mesenteroides AJ13, Pediococcus halophilus AJ22, Lactobacillus sakei AJ29, and Pediococcus pentosaceus AJ35. The strains AJ06, AJ22, AJ29 exhibited high tolerance to simulated gastric and intestinal juices and were able to produce bile salt hydrolase on MRS agar plates supplemented with taurocholic acid and/or taurodeoxycholic acid. The strains AJ22 and AJ29, which demonstrated high adherence to Caco-2 cells and resistance to various antibiotics, effectively inhibited the growth of food-borne pathogens by the production of antimicrobial substances. These strains did not show ${\alpha}-$ or ${\beta}$-haemolysis on blood agar. Furthermore, biogenic amines in MRS broth containing the precursor amino acids were not mutagenic in Salmonella Typhimurium TA98 and TA100.

Preparation of $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ by Bacillus sp. ${\beta}-mannanase$ and Growth Activity to Intestinal Bacteria (Bacillus sp.유래 ${\beta}-mannanase$에 의한 $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ 조제 및 장내세균에 대한 생육활성)

  • Kim, Sang-Woo;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.379-383
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    • 2004
  • For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.

Production of gamma-Aminobutyric Acid (GABA) by Lactobacillus plantarum subsp. plantarum B-134 Isolated from Makgeolli, Traditional Korean Rice Wine (한국전통주인 막걸리로부터 분리한 Lactobacillus plantarum subsp. plantarum B-134의 gamma-aminobutyric acid (GABA)의 생산)

  • Lee, Hyun-Ju;Son, Jae-Young;Lee, Sang-Jae;Lee, Han-Seung;Lee, Bae-Jin;Choi, In-Soon;Sohn, Jae Hak
    • Journal of Life Science
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    • v.27 no.5
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    • pp.567-574
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    • 2017
  • This study is to isolate and identify ${\gamma}$-amino butyric acid (GABA) producing lactic acid bacteria (LAB) from Makgeolii, traditional Korean rice wine and then establish the optimal culture conditions for GABA production. Sixty four LAB from Makgeolli were isolated according to the characteristics of the shape and color of the colony grown on MRS agar plate. The GABA production of the isolated strain cultured in MRS broth contained 1% MSG (mono-sodium glutamate) were determined and evaluated by TLC and HPLC analysis. Strain B-134 was selected for highest GABA production. From the analysis of 16S rRNA and glutamate decarboxylase B (gadB) gene sequences, strain B-134 was tentatively identified as a Lactobacillus plantarum subsp. plantarum B-134. Effects of culture parameters, including glutamic acid level, culture temperature, NaCl level, and pH on GABA production were investigated for culture optimization. The optimum culture condition for GABA production by B-134 were culture temperature of $37^{\circ}C$, pH of 5.7, NaCl content of 0% (w/v) and MSG content of 3% (w/v), which produced 25 mM of GABA during cultivation time of 48 hr. From these results, strain B-134 is expected to be utilized as useful microorganisms for GABA-enriched health beneficial food.

Antibacterial Activity and Characteristics of Bacteriocin Produced by Lactobacillus plantarum LMG 7945 (Lactobacillus plantarum LMG 7945가 생산하는 bacteriocin의 항균력과 그 특성)

  • 김상현;이명숙;장동석
    • Journal of Food Hygiene and Safety
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    • v.10 no.2
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    • pp.65-71
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    • 1995
  • Bacteriocins from lactic acid bacteria have attracted much attention in recent years because of their useful worth in increasing safety and extending shelf life of foods. These substances show an inhibitory effect against some food spoilage bacteria and food-borne pathogens. The inhibitory effect fo the bacteriocin produces by lactic acid bacteria against Listeria monocytogenes(L. monocytogenes) was examined in this study. The culture supernatants of 5 kinds of bacteria among the 10 kinds of testes lactic acid bacteria had the inhibitory activity against Listeria sp., various Gram positive and Gram negative bacteria. Bacteriocin produced by Lactobacillus plantarum(Lact. plantarum) LMG 7945 was the most active toward L. monocytogenes. Bacteriocin production of the Lact. plantarum LMG 7945 cultured on MRS broth was increased late logarithmic phase over early stationary phase. This bacteriocin was stable at heat treatment and acidic pH relatively; The activity was retained after heating at 121$^{\circ}C$ for 15min and was active in the pH range of 2~4 but was lost above pH 5.

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동물의 장에서 분리한 Enterococcus sp.의 특성 및 분말화

  • Park, Chong-Jin;Pyeon, Jeong-Soo;Cho, Yun-Kyung;Hong, Seung-Suh;Lee, Hyun-Soo
    • Microbiology and Biotechnology Letters
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    • v.24 no.4
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    • pp.393-398
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    • 1996
  • In order to develop a lactic acid bacterial powder which can be used as a probiotic for human and animal, a lactic acid bacteria which has high resistance against low pH and ox-gall, and shows a good growth inhibition against E. coli, was isolated from an animal intestine and characterized. The isolated strain was identified as Enterococcus faecium. It had more than 90% of survival at low pH for 2 hours and almost 100% of survival in the presence of 0.3% ox-gall. When co-cultured with E. coli in MRS broth, all of the E. coli cells were killed within 24 hours. The final powdered product of the isolated strain was manufactured after a freeze drying process using an industrial media, and then checked its stability. Its storage stability was 80% for 11 months at 18$\circ$C.

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Production of ${\gamma}-Aminobutyric$ Acid (GABA) by Lactobacillus buchneri Isolated from Kimchi and its Neuroprotective Effect on Neuronal Cells

  • Cho, Yu-Ran;Chang, Ji-Yoon;Chang, Hae-Choon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.104-109
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    • 2007
  • Lactic acid bacteria that accumulated ${\gamma}-aminobutyric$ acid (GABA) in culture medium were screened to identify strains with high GAB A-producing ability. One strain, MS, which was isolated from kimchi, showed the highest GABA-producing ability among the screened strains. MS was identified as Lactobacillus buchneri based on Gram-staining, metabolic characteristics, and 16S rDNA sequence determination, Optimum culture conditions for GABA production were determined: MRS broth containing 5% MSG, 1% NaCl, and 1% glucose, at an initial pH of 5.0, the incubation temperature at $30^{\circ}C$ for 36 h. Under these conditions, MS produced GABA at a concentration of 251 mM with a 94% GABA conversion rate. Moreover, culture extracts of Lb. buchneri MS partially or completely protected neuronal cells against neurotoxicantinduced cell death.

Biological Function of Lactoferrin in Milk

  • Kei-Ichi, Shimazaki
    • 한국유가공학회:학술대회논문집
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    • 2002.04a
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    • pp.37-42
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    • 2002
  • Lactoferrin is an iron-binding glycoprotein and its bacteriostatic and bactericidal effects on Gram-positive and Gram-negative bacteria have been well-known. However, certain kind of lactic acid bacteria are resistant against its antibacterial effects. Moreover, it is reported that lactoferrin promotes the growth of bifidobacteria by in vitro and in vivo experiments. In this experiment, lactoferrin-binding protein was found both in the membrane and cytosolic franctions of Bifidobacterium. Bifidobacterium was grown in anaerobic conditions in MRS broth containing cysteine, gathered by centrifugation and processed by sonication. The lactoferrin-binding proteins on the PVDF-membrane transferred after SDS-PAGE were detected by far-western method using biotinylated lactoferrin and streptavidin-labeled horse radish peroxidase. Observation in growth effects of lactoferrin on Bifidobacterium suggested that there is a relation between the presence of lactoferrin-binding proteins on the cells and their growth.

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