• Title/Summary/Keyword: identification of lactic acid bacteria

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Antibacterial Effects of Lactococcus lactis HK-9 Isolated from Feces of a New Born Infant (신생아 태변에서 젖산세균인 Lactococcus lactis HK-9의 분리 및 항균활성)

  • Baek, Hyun;Ahn, Hye-Ran;Cho, Yun-Seok;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.127-133
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    • 2010
  • The purpose of this work was to investigate the antibacterial activity derived from a lactic acid bacterium, Lactococcus lactis HK-9, isolated from the feces of a 2-day newborn infant. We characterized the physiological and biochemical properties of this strain. Both the BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were utilized for identification, and the strain was assigned to the Lactococcus lactis species, designated as L. lactis HK-9, and registered in GenBank as [GU936712]. We monitored growth rate, production of lactic acid and acetic acid as metabolites, and pH during growth. The maximum concentrations of lactic acid and acetic acid reached 495.6 mM and 104.3 mM, respectively, and the initial pH of the cultures decreased from 7.0 to 4.1 after incubating for 60 h. HPLC was used to confirm the production of lactic acid and acetic acid. Significant antibacterial activity of the concentrated supernatant was demonstrated against Gram-positive (e.g., Staphylococcus aureus, Enterococcus faecalis, Listeria monocytogenes, MRSA) and Gram-negative (e.g., Escherichia coli, Salmonella enteritidis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Shigella sonnei) bacteria by the plate diffusion method. The antibacterial activity was sensitive to protease, and the molecular weight of the presumed bacteriocin molecule was estimated to be about 4 kDa by tricine-SDS-PAGE.

Angiotensin-I-Converting Enzyme Inhibitory Peptides in Goat Milk Fermented by Lactic Acid Bacteria Isolated from Fermented Food and Breast Milk

  • Rubak, Yuliana Tandi;Nuraida, Lilis;Iswantini, Dyah;Prangdimurti, Endang
    • Food Science of Animal Resources
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    • v.42 no.1
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    • pp.46-60
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    • 2022
  • In this study, angiotensin-I-converting enzyme inhibitory (ACEI) activity was evaluated in fermented goat milk fermented by lactic acid bacteria (LAB) from fermented foods and breast milk. Furthermore, the potential for ACEI peptides was identified in fermented goat milk with the highest ACEI activity. The proteolytic specificity of LAB was also evaluated. The 2% isolate was inoculated into reconstituted goat milk (11%, w/v), then incubated at 37℃ until pH 4.6 was reached. The supernatant produced by centrifugation was analyzed for ACEI activity and total peptide. Viable cell counts of LAB and titratable acidity were also evaluated after fermentation. Peptide identification was carried out using nano liquid chromatography mass spectrometry (LC-MS/MS), and potential as an ACEI peptide was carried out based on a literature review. The result revealed that ACEI activity was produced in all samples (20.44%-60.33%). Fermented goat milk of Lc. lactis ssp. lactis BD17 produced the highest ACEI activity (60.33%; IC50 0.297±0.10 mg/mL) after 48 h incubation, viable cell counts >8 Log CFU/mL, and peptide content of 4.037±0.27/mL. A total of 261 peptides were released, predominantly derived from casein (93%). The proteolytic specificity of Lc. lactis ssp. lactis BD17 through cleavage on the amino acid tyrosine, leucine, glutamic acid, and proline. A total of 21 peptides were identified as ACEI peptides. This study showed that one of the isolates from fermented food, namely Lc. lactis ssp. lactis BD17, has the potential as a starter culture for the production of fermented goat milk which has functional properties as a source of antihypertensive peptides.

Isolation and Identification of Lactic Acid Bacteria for Preparation of Probiotics (생균제용 유산균의 분리 및 동정)

  • 이진규;김운태;이준호;유재홍;신원철
    • Microbiology and Biotechnology Letters
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    • v.19 no.5
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    • pp.429-432
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    • 1991
  • In order to prepare probiotics, a strain which has resistant against acid, bile salts and antibiotics was isolated from infant and anirnat feces. Among these isolated strains, TK- 19 showed a strong acid tolerance and growth at 500 $\mu g$/ml of Na-deoxycholate. Also strain TK-19 was resistant to kanarnycin (30 $\mu g$/ml), streptomycin (10 $\mu g$/ml), nisin (50 $\mu g$/ml) and tetracycline (10 $\mu g$/ml). The isolated strain TK-19 was identified as Streptococcus faeciam, and was named Streptococcus faeciurn TK-19.

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Short-Term Changes in Gut Microflora and Intestinal Epithelium in X-Ray Exposed Mice

  • Tsujiguchi, Takakiyo;Yamaguchi, Masaru;Yamanouchi, Kanako
    • Journal of Radiation Protection and Research
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    • v.45 no.4
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    • pp.163-170
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    • 2020
  • Background: Gut microflora contributes to the nutritional metabolism of the host and to strengthen its immune system. However, if the intestinal barrier function of the living body is destroyed by radiation exposure, the intestinal bacteria harm the health of the host and cause sepsis. Therefore, this study aims to trace short-term radiation-induced changes in the mouse gut microflora-dominant bacterial genus, and analyze the degree of intestinal epithelial damage. Materials and Methods: Mice were irradiated with 0, 2, 4, 8 Gy X-rays, and the gut microflora and intestinal epithelial changes were analyzed 72 hours later. Five representative genera of Actinobacteria, Firmicutes, and Bacteroidetes were analyzed in fecal samples, and the intestine was pathologically analyzed by Hematoxylin-Eosin and Alcian blue staining. In addition, DNA fragmentation was evaluated by the TdT-mediated dUTP nick-end labeling (TUNEL) assay. Results and Discussion: The small intestine showed shortened villi and reduced number of goblet cells upon 8 Gy irradiation. The large intestine epithelium showed no significant morphological changes, but the number of goblet cells were reduced in a radiation dose-dependent manner. Moreover, the small intestinal epithelium of 8 Gy-irradiated mice showed significant DNA damaged, whereas the large intestine epithelium was damaged in a dose-dependent manner. Overall, the large intestine epithelium showed less recovery potential upon radiation exposure than the small intestinal epithelium. Analysis of the intestinal flora revealed fluctuations in lactic acid bacteria excretion after irradiation regardless of the morphological changes of intestinal epithelium. Altogether, it became clear that radiation exposure could cause an immediate change of their excretion. Conclusion: This study revealed changes in the intestinal epithelium and intestinal microbiota that may pave the way for the identification of novel biomarkers of radiation-induced gastrointestinal disorders and develop new therapeutic strategies to treat patients with acute radiation syndrome.

Identification of the Predominant Species of Bacillus, Staphylococcus, and Lactic Acid Bacteria in Nuruk, a Korean Starter Culture (배양법을 이용한 누룩 발효 관련 Bacillus 속, Staphylococcus 속 세균 및 유산균의 우점종 확인)

  • Saeyoung Seo;Do-Won Jeong;Jong-Hoon Lee
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.93-98
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    • 2023
  • Nuruk is a starter culture of Korea manufactured by spontaneous fermentation of grains. We isolated bacteria of the genera Bacillus and Staphylococcus, and lactic acid bacteria (LAB) from eight commercial nuruk samples collected from four districts of Korea using selective agar media and identified them based current taxonomic standards. Bacillus was detected in all samples, but Staphylococcus or LAB were not detected in three samples. In seven samples, except one sample scored the highest cell number of LAB, Bacillus and Staphylococcus were counted as the highest and the lowest numbers, respectively. Six species of Bacillus were identified, and B. subtilis, B. velezensis, and B. licheniformis were predominant species. Nine species of coagulase-negative Staphylococcus were identified, and the predominance of S. pseudoxylosus and S. saprophyticus was confirmed. Ten species of LAB including Enterococcus, Lactobacillus and close relatives, Pediococcus, and Weissella were identified. P. pentosaceus was identified as the predominant species.

A Study on Microbial Aspects of Korean Human Milk by Collection Methods (수집방법에 따른 한국인 모유의 미생물 분포에 관한 연구)

  • 이조윤;배형철;남명수
    • Food Science of Animal Resources
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    • v.23 no.3
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    • pp.269-277
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    • 2003
  • This study was carried out to evaluate the safety of Korean human milk. The microorganisms were identified from human milk of 149 healthy mothers by two collection methods, hand and pump expression. The means of total bacterial counts were 2.33x10$^4$ cfu/mL on the samples collected by the pump expression and 7.83xl0$^3$ cfu/mL on those collected by the hand expression. Therefore, the total bacterial counts of pump expression samples was 9.80xl0$^2$∼3.06x10$^4$ cfu/mL more than that of hand expression samples. The coliform counts of pump expression was 9.36xl0$^3$∼8.57xl0$^4$ cfu/mL more than that of hand expression. However, there was any significant differences of the lactic acid bacterial counts between the two samples collected by each methods. 100 strains of 5 patterns of total bacterial counts were isolated based on the morphology of colony in the standard plate count agar. 13 species were identified among the isolated strains. The dominant species in Korean human milk were Staphylococcus which 7 subspecies identified(81% in the rate of total bacteria, 1.07x10$^4$ cfu/mL). Other species identified were Micrococcus, Bacillus, Providencia, Pseudomonas, Yersinia and Acinetobacter. 36 strains of 6 patterns of lactic acid bacterial counts were isolated based on morphology of colony in the BCP agar. 7 species were identified among the isolated strains. The dominant species of lactic acid bacteria in Korean human milk were Lactobacillus brevis(50.9% in the rate of lactic acid bacteria, 4.72xl0$^4$ cfu/mL). Others species identified(49.1% lactic acid bacteria) were Lactobacillus curvatus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus acidophilus, Leuconostic lactis and Streptococcus salivarius subsp. thermophilus.

Isolation of Bacteriocin-Producing Lactococcus sp.HY 449 and Its Antimicrobial Characteristics (Bacteriocin을 생산하는 Lactococcus sp. HY 449의 분리와 항균 특성)

  • Kim, Sang-Kyo;Lee, Sang-Jun;Baek, Young-Jin;Park, Yun-Hee
    • Microbiology and Biotechnology Letters
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    • v.22 no.3
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    • pp.259-265
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    • 1994
  • A bacteriocin-producing lactic acid bacteria was isolated from contaminated milk products, which was identified by using the API50 CH kit as Lactococcus lactis subsp. lactis with reliability of 98%. Fatty and analysia of the cell membrane showed that this strain contained same fatty acids profiles as type strain, Lactococcus lactis subsp. lactis ATCC 19435. The bacteriocin of Lactococcus sp. HY 449 showed relatively wide range of inhibition spectrum against gram positive and some gram negative bacteria such as Escherichia coli and maintained the inhibitory activity between pH2.0 and pH9.0 The thermostability of this bacteriocin was higher in acidic solution than in distilled water and was stable at 60$\circ $C for 1 hour.

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Characteristics of Takju (a Cloudy Korean Rice Wine) Prepared with Nuruk (a Traditional Korean Rice Wine Fermentation Starter), and Identification of Lactic Acid Bacteria in Nuruk (시판 전통누룩의 젖산균 분리동정과 제조단양주의 품질특성)

  • Park, Ji-Hee;Chung, Chang-Ho
    • Korean Journal of Food Science and Technology
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    • v.46 no.2
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    • pp.153-164
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    • 2014
  • Five takju prepared using four types of nuruk (a traditional Korean fermentation starter made of malted wheat; non-cooked, naturally inoculated) labeled SH, SS, JJ, and SJ, and one type of koji (cooked, inoculated with an inoculum) labeled MN, were compared. Titratable acidity, pH, sugars, ethanol, amino acids, organic acids, and microbial changes in samples were measured, and the sensory properties were evaluated. Titratable acidity, alcohol, and organic acid content increased as sugar contents decreased. The overall ethanol concentration of all takju increased over time, reaching a maximum of 13.08-14.96% (w/v) at 7-21 days. The total amino acid contents of takju prepared with nuruk, except for one (SJ), were higher than the takju prepared with koji (MN). Lactic acid bacteria were also isolated from the starters. Sequence analysis of 16S rRNA genes (500 - 600 bp) of 223 isolates revealed that the major strains were in the genera of Leuconostoc, Weissella, Pediococcus, and Lactobacillus.

Characterization of Cholesterol Lowering Lactic Acid Bacteria Isolated from Palm Wine and Maize Beer and Assessment of Their Use in the Production of Probiotic Papaya Juice

  • Bertrand Tatsinkou Fossi;Dickson Ebwelle Ekabe;Liliane Laure Toukam Tatsinkou;Rene Bilingwe Ayiseh;Frederic Tavea;Pierre Michel Jazet
    • Microbiology and Biotechnology Letters
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    • v.51 no.2
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    • pp.191-202
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    • 2023
  • Elevated serum cholesterol is a main risk factor for heart disorders. Most probiotic products administered to lower cholesterol are dairy products which are not suitable for lactose-intolerant individuals. In this study, we assessed the cholesterol-lowering efficacy of LAB isolated from traditionally fermented drinks in diet-induced rats and determine their efficacy in the production of non-dairy, probiotic formulations using papaya juice. LAB were isolated from palm wine and corn beer on MRS agar using a pour-plate technique. Identification was carried out using 16S rRNA gene sequencing. A hypercholesterolemia model in which diet-induced Wistar albino rats were assigned into four groups was established. Oral gavage was carried out for 30 days. On the 31st day, the rats were dissected and the serum lipid profile was analyzed using biochemical kits. A 106 cfu/ml of a 24-h-old culture of selected lactobacilli was used to inoculate papaya juice and incubated at 37℃. Microbial and chemical changes were assessed during papaya fermentation and after four weeks of cold storage. Two selected isolates (Pw1 and Cb4) had in vitro cholesterol reduction of > 80%. These two isolates lowered lipid profile (triglyceride, total cholesterol, LDL-c) significantly, and increased HDL-c levels (p < 0.5) in the rat sera. Phylogenetic analysis showed that Pw1 was 98.86% similar to Limosilactobacillus fermentum, while Cb4 was 99.54% similar to Enteroccocus faecium. Both strains fermented papaya juice with cell viability reaching 8.92 × 108 cfu/ml and 25.3 × 108 cfu/ml respectively, and were still viable after 4 weeks of cold storage.

Effect of Low Salt Concentrations on Microbial Changes During Kimchi Fermentation Monitored by PCR-DGGE and Their Sensory Acceptance

  • Ahmadsah, Lenny S. F.;Min, Sung-Gi;Han, Seon-Kyeong;Hong, Yeun;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2049-2057
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    • 2015
  • Various salt concentrations (1.0%, 1.3%, 1.6%, 1.9%, and 2.1% labeled as sample A, B, C, D, and E, respectively) were investigated for microbial diversity, identification of Lactic Acid Bacteria (LAB) in salted kimchi cabbage, prepared under laboratory conditions. These samples were stored at 4°C for 5 weeks in proper aluminum-metalized pouch packaging with calcium hydroxide gas absorber. A culture-independent method known as polymerase chain reaction - denaturing gradient gel electrophoresis was carried out to identify LAB distributions among various salt concentration samples that had identified 2 Weissella (W. confusa and W. soli), 1 Lactobacillus (Lb. sakei), and 3 Leuconostoc (Lc. mesenteroides, Lc. lactis, and Lc. gelidum) in the overall kimchi samples. The pH, titratable acidity, viable cell counts, and coliform counts were not affected by salt variations. In order to assess sensory acceptance, the conducted sensory evaluation using a 9-point hedonic scale had revealed that samples with 1.3% salt concentration (lower than the manufacturer's regular salt concentration) was more preferred, indicating that the use of 1.3% salt concentration was acceptable in normal kimchi fermentation for its quality and safety. Despite similarities in pH, titratable acidity, viable cell counts, coliform counts, and LAB distributions among the various salt concentrations of kimchi samples, the sample with 1.3% salt concentration was shown to be the most preferred, indicating that this salt concentration was suitable in kimchi production in order to reduce salt intake through kimchi consumptions.