• Title/Summary/Keyword: 16S-rRNA

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Potential of Antifungal Lactic Acid Bacteria Isolated from Kimchi as Cheese Starters (김치 분리 항진균 유산균의 치즈 스타터로서 이용 가능성)

  • Oh, Hyun Hee;Huh, Chang Ki;Choi, Ha Nuel;Yang, Hee Sun;Bae, In Hyu;Lee, Jai Sung;Jeong, Yong Seob;Lee, Nam Keun;Jung, Hoo Kil
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.133-141
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    • 2013
  • This study was performed to identify the cheese starter potential of antifungal lactic acid bacteria isolated from Kimchi. Eight fungi were isolated from cheese or the cheese ripening room, and identified as Penicillium and Cladosporium by ITS-5.8S rDNA analysis. Twenty-two lactic acid bacteria species with antifungal activity were isolated from Kimchi, and identified as Lactobacillus and Pediococcus by 16S rRNA sequence analysis. Six lactic acid bacteria species were selected (L. sakei subsp. ALJ011, L. sakei subsp. ALI033, L. sakei subsp. ALGy039, P. pentosaceus ALJ015, P. pentosaceus ALJ024, and P. pentosaceus ALJ026) based on higher antifungal activity from the initial 22 species. Out of the six identified species, L. sakei subsp. ALI033 had the highest antifungal activity. For growth of the six lactic acid bacteria, optimal temperature and pH were $30{\sim}37^{\circ}C$ and 7.0, respectively. Proteolytic activities of the six lactic acid bacteria were almost as strong as the commercial strain Str. thermophilus Body-1. Coagulative activities of L. sakei subsp. ALI033, P. pentosaceus ALJ015, and P. pentosaceus ALJ024 were higher than those of L. sakei subsp. ALJ011, L. sakei subsp. ALGy039, and P. pentosaceus ALJ026. The acid resistance of L. sakei subsp. was higher than that of P. pentosaceus. The major organic acid component of the lactic acid bacteria culture medium was lactic acid.

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Optimization of Medium for the Carotenoid Production by Rhodobacter sphaeroides PS-24 Using Response Surface Methodology (반응 표면 분석법을 사용한 Rhodobacter sphaeroides PS-24 유래 carotenoid 생산 배지 최적화)

  • Bong, Ki-Moon;Kim, Kong-Min;Seo, Min-Kyoung;Han, Ji-Hee;Park, In-Chul;Lee, Chul-Won;Kim, Pyoung-Il
    • Korean Journal of Organic Agriculture
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    • v.25 no.1
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    • pp.135-148
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    • 2017
  • Response Surface Methodology (RSM), which is combining with Plackett-Burman design and Box-Behnken experimental design, was applied to optimize the ratios of the nutrient components for carotenoid production by Rhodobacter sphaeroides PS-24 in liquid state fermentation. Nine nutrient ingredients containing yeast extract, sodium acetate, NaCl, $K_2HPO_4$, $MgSO_4$, mono-sodium glutamate, $Na_2CO_3$, $NH_4Cl$ and $CaCl_2$ were finally selected for optimizing the medium composition based on their statistical significance and positive effects on carotenoid yield. Box-Behnken design was employed for further optimization of the selected nutrient components in order to increase carotenoid production. Based on the Box-Behnken assay data, the secondary order coefficient model was set up to investigate the relationship between the carotenoid productivity and nutrient ingredients. The important factors having influence on optimal medium constituents for carotenoid production by Rhodobacter sphaeroides PS-24 were determined as follows: yeast extract 1.23 g, sodium acetate 1 g, $NH_4Cl$ 1.75 g, NaCl 2.5 g, $K_2HPO_4$ 2 g, $MgSO_4$ 1.0 g, mono-sodium glutamate 7.5 g, $Na_2CO_3$ 3.71 g, $NH_4Cl$ 3.5g, $CaCl_2$ 0.01 g, per liter. Maximum carotenoid yield of 18.11 mg/L was measured by confirmatory experiment in liquid culture using 500 L fermenter.

Comparative Analysis of Gut Microbiota among Broiler Chickens, Pigs, and Cattle through Next-generation Sequencing (차세대염기서열 분석을 이용한 소, 돼지, 닭의 장내 미생물 군집 분석 및 비교)

  • Jeong, Ho Jin;Ha, Gwangsu;Shin, Su-Jin;Jeong, Su-Ji;Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1079-1087
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    • 2021
  • To analyze gut microbiota of livestock in Korea and compare taxonomic differences, we conducted 16S rRNA metagenomic analysis through next-generation sequencing. Fecal samples from broiler chickens, pigs, and cattle were collected from domestic feedlots randomly. α-diversity results showed that significant differences in estimated species richness estimates (Chao1 and ACE, Abundance-based coverage estimators) and species richness index (OUTs, Operational taxonomic units) were identified among the three groups. However, NPShannon, Shannon, and Simpson indices revealed that abundance and evenness of the species were statistically significant only for poultry (broiler chickens) and mammals (pigs and cattle). Firmicutes was the most predominant phylum in the three groups of fecal samples. Linear discriminant (LDA) effect size (LEfSe) analysis was conducted to reveal the ranking order of abundant taxa in each of the fecal samples. A size-effect over 2.0 on the logarithmic LDA score was used as a discriminative functional biomarker. As shown by the fecal analysis at the genus level, broiler chickens were characterized by the presence of Weissella and Lactobacillus, as well as pigs were characterized by the presence of provetella and cattele were characterized by the presence of Acinetobacter. A permutational multivariate analysis of variance (PERMANOVA) showed that differences of microbial clusters among three groups were significant at the confidence level. (p=0.001). This study provides basic data that could be useful in future research on microorganisms associated with performance growth, as well as in studies on the livestock gut microbiome to increase productivity in the domestic livestock industry.

Change in the composition and enzyme activity of culturable lactic acid bacteria in Nuruk during fermentation at different temperatures (온도를 달리한 누룩 발효 기간별 배양 유산균 변화 및 분리 유산균들의 효소 활성)

  • Nam, Kang;Lee, Nam Keun;Yum, Eun-Ji;Kim, Yong-Sik;Kim, Dae-Hyuk;Yeo, Soo-Hwan;Jeong, Yong-Seob
    • Food Science and Preservation
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    • v.22 no.6
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    • pp.920-925
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    • 2015
  • The microbial composition in Nuruk, a Korean cereal fermentation starter, is a critical factor for the quality and organoleptic properties of traditional alcoholic beverages. This study was aimed at monitoring the compositional change and enzyme activity of culturable lactic acid bacteria (LAB) in two types of Nuruk fermented at different temperatures. All culturable LAB were isolated at various time points (0, 3, 6, 10, 20, and 30 days) and identified by 16S rRNA sequencing. In traditional Nuruk type A (TN-A), which was fermented at $36^{\circ}C$, the population of total culturable LAB during the fermentation period was between $10^4$ and $10^5$ log CFU/mL. On the other hand, the LAB population in traditional Nuruk type B (TN-B) fermented at $45^{\circ}C$ (primary fermentation for 10 days) and $35^{\circ}C$ (secondary fermentation for 20 days) was $10^2$ log CFU/mL; however, these bacteria could not be detected after 6 days. Major LAB strains were identified in both Nuruk types: (1) from the MRS-culture of TN-A, Pediococcus pentosaceus at 3-30 days; (2) from MRS-culture of TN-B, P. pentosaceus at 3 days and Enterococcus hirae at 6 days. The protease activities of the dominant LAB isolated from the TN-A and TN-B cultures were within the ranges of 0.64~1.03 mg/mL and 0.74~0.81 mg/mL (tyrosine content), respectively, whereas the ${\alpha}$-amylase activities were 0.75~0.98 mg/mL and 0.78~0.79 mg/mL (amylose content), respectively.

Production and fermentation characteristics of seafood kimchi started with Leuconostoc mesenteriodes SK-1 isolated from octopus baechu kimchi (문어 배추김치에서 분리한 Leuconostoc mesenteroides SK-1을 이용한 수산물김치의 제조 및 발효 특성)

  • Jang, Mi-Soon;Jung, Ko-Eun;Yun, Jae-Ung;Nam, Ki-Ho
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.1050-1057
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    • 2016
  • This study was carried out to investigate the effectiveness of Leuconostoc mesenteroides isolated from octopus baechu kimchi as a potential starter for seafood kimchi. L. mesenteroides is lactic acid bacterium currently used as a starter for kimchi production. We selected the most effective L. mesenteroides strain from the 7 strains isolated from octopus baechu kimchi and, based on biochemical properties and 16S rRNA sequencing, identified the selected strain as L. mesenteroides SK-1. The SK-1 strain exhibited acid-tolerance, good survival capacity, and excellent dextran productivity. We investigated the effects the SK-1 of starter on seafood kimchi fermentation. Octopus baechu kimchi was fermented with L. mesenteroides SK-1 at $4^{\circ}C$ for 35 d. The decrease in pH and increase in acidity in octopus baechu kimchi fermented with the SK-1 starter occurred more quickly than that in the control kimchi indicating that. Octopus baechu kimchi with SK-1 starter has a relatively slow rate of increase in lactic acid production. As a result, octopus baechu kimchi prepared with L. mesenteroides SK-1 can be maintained at a suitable ripening degree over an extended period of time compared to that of the control kimchi, Moreover, the octopus baechu kimchi started with L. mesenteroides SK-1 has excellent sensory properties, including a refreshing taste, and a weak sour odor.

Selection and appropriate culture conditions of antagonistic bacterium Bacillus altitudinis HC7 against button mushroom cobweb disease caused by Cladobotryum mycophilum (양송이버섯 솜털곰팡이병균(Cladobotryum mycophilum)에 대한 길항미생물 Bacillus altitudinis HC7의 선발 및 적정 배양조건)

  • Chan-Jung Lee;Hye-Sung Park;Seong-Yeon Jo;Gi-Hong An;Ja-Yun Kim;Kang-Hyo Lee
    • Journal of Mushroom
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    • v.22 no.2
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    • pp.60-66
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    • 2024
  • This study was conducted to selection and investigate appropriate conditions for mass production of antagonistic microbes to control cobweb disease caused by Cladobotryum mycophilum. A grampositive bacterium was isolated from spent substrate of Agaricus bisporus and showed significant antagonistic activity against Cladobotryum mycophilum. The bacterium was identified as Bacillus altitudinis. based on the cultural, biochemical and physiological characteristics, and 16S rRNA sequence. The isolate is saprophytic, but not parasitic nor pathogenic to cultivated mushroom whereas it showed strong inhibitory effects against C. mycophilum cells in vitro. The control efficacy of B. altitudinis HC7 against cobweb disease of C. mycophilum was up to 78.2% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol system. To define the appropriate conditions for the mass production of the Bacillus altitudinis HC7, we have investigated appropriate culture conditions and effects of various nutrient source on the bacterial growth. The appropriate initial pH and temperature were determined as pH 6.0 and 30℃, respectively. The appropriate concentration of medium elements for the growth of pathogen inhibitor bacterium(Bacillus altitudinis HC7) was determined as follows: 3.0% soluble startch, 10% soytone, 1.0% (NH4)2HPO4, 1.0 mmol KCl, and 0.5% L-asparagine.

Biogeochemical Remediation of Cr(VI)-Contaminated Groundwater using MMPH-0 (Enterobacter aerogenes) (MMPH-0 (Enterobacter aerogenes)에 의한 6가 크롬 오염 지하수의 생지화학적 정화)

  • Seo, Hyun-Hee;Rhee, Sung-Keun;Kim, Kang-Joo;Park, Eun-Gyu;Kim, Yeong-Kyoo;Chon, Chul-Min;Moon, Ji-Won;Roh, Yul
    • Economic and Environmental Geology
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    • v.45 no.2
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    • pp.105-119
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    • 2012
  • Indigenous bacteria isolated from contaminated sites play important roles to remediate contaminated groundwater. Chromium has the most stable oxidation states. Cr(VI) is toxic, carcinogenic, and mobile, but Cr(III) is less toxic and immobile. In this study, indigenous microorganism (MMPH-0) was enriched from Cr(VI) contaminated groundwater, and identified by 16S rRNA gene analysis. Using MMPH-0, the effect of stimulating with e-donors (glucose, lactate, acetate, and no e-donor control), respiration conditions, biomass, tolerance, and geochemical changes on Cr(VI) reduction were investigated in batch experiments for 4 weeks. The changes of Cr(VI) concentration and geochemical conditions were monitored using UV-vis-spectrophotometer and Eh-pH meter. And the morphological and chemical characteristics of MMPH-0 and precipitates in the effluents were characterized by TEM-EDS and SEM-EDS analyses. MMPH-0 (Enterobacter aerogenes) was able to tolerate up to 2000 mg/L Cr(VI) and reduce Cr(VI) under aerobic and anaerobic conditions. MMPH-0 performed faster and higher efficiency of Cr(VI) reduction with electron donors (over 70% after 1 week with e-donor, 10-20% after 4 weeks without e-donor). The changes of Eh-pH in effluents showing the tendency from oxidizing to reducing condition and a bit of acidic change in pH due to microbial oxidation of organic matters donating electrons and protons suggested the roles of MMPH-0 on Cr(VI) in the contaminated water catalyzing to transit geochemical stable zone for more stable $Cr(OH)_3$ or Cr(III) precipitates. TEM/SEM-EDS analyses of MMPH-0 and precipitates indicate direct and indirect Cr(VI) reduction: extracellular polymers capturing Cr component outside cells. These results suggested diverse indigenous bacteria and their biogeochemical reactions might enhance more effective and feasible remediation technology of redox sensitive heavy metals in metal-contaminated in groundwater.

Production of Poly-3-hydroxybutyrate from Xylose by Bacillus megaterium J-65 (Bacillus megaterium J-65에 의한 xylose로부터 poly-3-hydroxybutyrate 생산)

  • Jun, Hong-Ki;Jin, Young-Hi;Kim, Hae-Nam;Kim, Yun-Tae;Kim, Sam-Woong;Baik, Hyung-Suk
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1625-1630
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    • 2008
  • A microorganism capable of producing high level of poly-3-hydoxybutyrate (PHB) from xylose was isolated from soil. The isolated strain J-65 was identified as Bacillus megaterium based on the morphological, biochemical and molecular biological characteristics. The optimum temperature and pH for the growth of B. megaterium J-65 were $37^{\circ}C$ and 8.0, respectively. The optimum medium composition for the cell growth was 2% xylose, 0.25% $(NH_4)_2SO_4$, 0.3% $Na_2HPO_4{\cdot}12H_2O$, and 0.1% $KH_2PO_4$. The optimum condition for PHB accumulation was same to the optimum condition for cell growth. Copolymer of ${\beta}$-hydroxybutyric and ${\beta}$-hydroxyvaleric acid was produced when propionic acid was added to shake flasks containing 20 g/l of xylose. Fermenter culture was carried out to produce the high concentration of PHB. In batch culture, cell mass was 9.82 g/l and PHB content was 35% of dry cell weight. PHB produced by B. megaterium J-65 was identified as homopolymer of 3-hydoxybutyric acid by GC and NMR.

Treatment of $Smilax$ $china$ L. Root Extract for Improvement of Storage Stability of $Mang-gae$ Rice Cake (망개떡의 저장성 향상을 위한 청미래덩굴 뿌리 추출물의 첨가)

  • Ko, Yu-Jin;Kim, Jin-Yong;Kim, Eun-Jung;Kim, Eun-Ja;Seol, Hui-Gyeong;Park, Geun-Hye;Chung, Gwon-Yong;Ryu, Chung-Ho
    • Food Science and Preservation
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    • v.19 no.2
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    • pp.167-172
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    • 2012
  • The antimicrobial activities of $Smilax$ $china$ L. against spoilage bacteria isolated from $Mang-gae$ rice cake were investigated and the storage stability of the $Mang-gae$ rice cake was enhanced. Spoilage bacteria, which cause $Mang-gae$ rice cake to rot, were isolated from commercial $Mang-gae$ rice cake, and most of the isolated strains were identified as $Bacillus$ sp. After the leaves, roots, shoots, and stalks of the $Smilax$ $china$ L. were extracted using 50% ethanol as the solvent, their antimicrobial activities were investigated using the paper disc method by treating them with 50 ${\mu}L$ of $Bacillus$ $cereus$, which is known as a major pathogenic micro-organism in foods that contain starch, as the test organism. The antimicrobial activities of the extracts were compared according to the size of the clear zones around the paper discs. The root extract showed significant antimicrobial activities. When red beans, which are used as stuffing for $Mang-gae$ rice cake, were treated with the root extract of the $Smilax$ $china$ L., the viable cell count of the $Mang-gae$ rice cake was 5.04 Log CFU/g after 48-hr storage, and the cake showed significantly slower growth of bacteria than with commercial products. These results show that treatment of red beans with $Smilax$ $china$ root extract could improve the storage stability of $Mang-gae$ rice cake.

Evaluation of Peptide Nucleic Acid Probe-Based Fluorescence In Situ Hybridization for the Detection of Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria in Clinical Respiratory Specimens (임상 객담검체에서 Peptide Nucleic Acid Probe를 이용한 결핵과 비결핵 항산균의 구분)

  • Lee, Seung Hee;Kim, Shine Young;Kim, Hyung Hoi;Lee, Eun Yup;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.18 no.2
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    • pp.37-43
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    • 2015
  • Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.