• 제목/요약/키워드: 16S-rRNA

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Physiological and Biochemical Characterization of Bacillus spp. from Polychaete, Perinereis aibuhitensis (갯지렁이(Perinereis aibuhitensis)에서 분리한 Bacillus spp.의 생리생화학적 특성 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Journal of Life Science
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    • v.23 no.3
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    • pp.415-425
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    • 2013
  • This study compared the characteristics of five Bacillus strains capable of aerobic and anaerobic growth, CBW3, CBW4, CBW9, CBW14 and EBW10. They were isolated and selected from a polychaete, Perinereis aibuhitensis, which is known as a good degrader of organic compounds in marine wetland. Based on a 16S rRNA sequence, CBW3 and CBW14 were found to share more than 99.8% similarity with B. nanhaiensis, B. arsenicus and B. barbaricus. CBW4, CBW9 and EBW10 shared 92.7%, 99.8%, and 99.8% similarity with B. anthracis, B. algicoa and B. thuringiensis, respectively. The temperature, salinity, and pH ranges of the cell growth of the Bacillus strains were $4-45^{\circ}C$, 0-17%, and pH 5-pH 9, respectively. All Bacillus strains were found to exhibit enzyme activities for the degradation of casein and starch. Notably, strain EBW10 exhibited the enzyme activities for all the tested macromolecules, DNA, casein, starch, cellulose, and four kinds of Tweens, which suggests the possibility that it had protease, amylase, cellulose, and lipase. All five Bacillus strains had alkaline phosphatase activities, and the strains CBW3, CBW4, and EBW10 also had acid phospatase. Strains CBW3 and EBW10 exhibited the enzyme activities both for esterase (C4) and esterase lipase (C8). The analysis of fatty acids revealed that in all strains, major fatty acids were anteiso $C_{15:0}$ and iso $C_{15:0}$.

Enhanced biological effect of fermented soy-powder milk with Lactobacillus brevis increasing in γ-aminobutyric acid and isoflavone aglycone contents (가바와 비당체 이소플라본이 증가된 Lactobacillus brevis 발효 콩-분말 두유의 생리활성 증진 효과)

  • Hwang, Chung Eun;Kim, Su Cheol;Lee, Jin Hwan;Hong, Su Young;Cho, Kye Man
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.245-255
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    • 2018
  • The research was aimed to analyze the functional constituents (GABA and isoflavone), radical (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and hydroxyl) scavenging activities and enzyme (${\alpha}-glucosidase$ and lipase) inhibitory effects of soypowder milk (SPM) and fermented soy-powder milk (FSPM) with varied Lactobacillus brevis. Ten ${\gamma}-aminobutyric$ acid (GABA) producing lactic acid bacteria that showed 96-99% similarity with L. brevis, according to 16S rRNA gene sequence analysis, were isolated from fermented kimchi. The conversion rates of GABA were obtained 66.96-93.51, 63.76-84.58, and 57.05-69.75% in monosodium glutamate, gluten and soy protein, respectively. The levels of pH and glutamic acid of FSPM were found lower than those of SPM, but the acidity and GABA contents were higher. The GABA conversion rate of FSPM with BMK484 strain was attained the highest 69.97%. The contents of isoflavone glycoside ($1290.93{\mu}g/g$) was higher in SPM, but the content of isoflavone aglycone ($287.27-501.9{\mu}g/g$) was higher in FSPM. The levels of isoflavone aglycone such as daidzein, glycitein and genistein, were found as the highest 240.2, 61.24 and $200.45{\mu}g/g$, respectively, when FSPM was made with BMK484 strain. The DPPH, ABTS and hydroxyl radical scavenging and ${\alpha}-glucosidase$ and pancreatic lipase inhibitory activities of FSPM made with BMK484 strain were the relatively higher 60.31, 88.10, 61.25, 52.71, and 39.37%, respectively. Therefore, the L. brevis can be used as a material capable of simultaneously enhanced GABA and isoflavone aglycone in FSPM.

Potential probiotics activity of Bacillus spp. from traditional soybean pastes and fermentation characteristics of Cheonggukjang (전통장류유래 Bacillus spp.의 프로바이오틱스 활성과 청국장 발효 특성)

  • Ryu, Myeong Seon;Yang, Hee-Jong;Kim, Jin Won;Jeong, Su-Ji;Jeong, Seong-Yeop;Eom, Jeong-Seon;Jeong, Do-Youn
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1168-1179
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    • 2017
  • In this study, we tried to screen the Bacillus strain having safety probability by isolation of strains from traditional fermented food, measurement of probiotic properties, and the fermentative characteristics of Cheonggukjang. We isolated 400 Bacillus-like isolates from traditional fermented foods. Selected strains examined on the prevalent characteristic such as extracellular enzyme and antibacterial activities, and their safety probability was confirmed by biogenic amine productivity, hemolytic, and harmful substances and enzyme productivity. We selected the 5 strains by analysis of biogenic amine, antibacterial and B. cereus toxic associated gene. Five selected strains were examined on cell surface hydrophobicity, and bile and acid tolerance, and we selected the SRCM100730 as the final strain. SRCM100730 was confirmed B. amyloliquefaciens by 16S rRNA sequencing, and named the B. amyloloquefaciens SRCM100730 (KCCM11966P). Finally, we manufactured Cheonggukjang using SRCM100730 for confirmation of fermentation properties. Manufactured Cheonggukjang did not contain B. cereus, and showed that ${\gamma}$-PGA and extracellular enzyme activities were superior to commercial Chunggukjang. Amino nitrogen content was 544.02 mg% and 26 free amino acid were detected, and the bitterness-related amino acid content was lower than commercial Cheonggukjang. Especially, the amount of GABA was 3 fold higher than commercial Cheonggukjang. These results suggest that SRCM100730 have high availability in commercial probiotics market and fermented food industry.

Antifungal activity against cheese fungi by lactic acid bacteria isolated from kimchi (김치 분리 유산균의 치즈 곰팡이 항진균 활성)

  • Choi, Ha Nuel;Oh, Hyun Hee;Yang, Hee Sun;Huh, Chang Ki;Bae, In Hyu;Lee, Jai Sung;Jeong, Yong Seob;Jeong, Eun Jeong;Jung, Hoo Kil
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.727-734
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    • 2013
  • The antifungal activity against cheese fungi by lactic acid bacteria isolated from kimchi was investigated. Eight fungi were isolated from cheese in the cheese ripening room. Two of them were identified as Penicillium and Cladosporium via ITS-5.8S rDNA analysis. Twenty-two species of lactic acid bacteria with antifungal activity were isolated from kimchi. Two of them were identified as Lactobacillus and Pediococcus via 16S rRNA sequence analysis. Of the 22 lactic acid bacteria species, six were selected (L. sakei subsp. ALJ011, L. sakei subsp. ALI033, L. sakei subsp. ALGy039, P. pentosaceus ALJ015, P. pentosaceus ALJ024 and P. pentosaceus ALJ026) due to their higher activity against the eight fungi isolated from cheese in the cheese ripening room; and among the six species, the P. pentosaceus ALJ015 and P. pentosaceus ALJ024 isolates from the Jeonju area kimchi and the L. sakei subsp. ALI033 isolate from the Iimsil area kimchi had higher antifungal activity than the other lactic acid bacteria. The minimum inhibitory concentration (MIC) of L. sakei subsp. ALI033 against the eight fungi isolated from cheese in the cheese ripening room was $62.5{\mu}g/mL$.

Characterization of Streptomyces Species Causing Potato Scab in Korea: Distribution, Taxonomy, and Pathogenicity

  • Lim, Chun-Keun;Park, Duck-Hwan;Kim, Jeom-Soon;Cho, Jun-Mo;Kwon, Soon-Wo;Hur, Jang-Hyun
    • The Plant Pathology Journal
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    • v.19 no.1
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    • pp.13-18
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    • 2003
  • From 1996 to 1999, potato-growing areas in Korea were surveyed for identification and distribution of potato scab pathogens. Potato scab was widely distributed in the mass cultivation areas, especially in Jriu island, southern areas of Chonnam and Gyounggi provinces, and the alpine area of Gangwon province. Jeju island was the most affected area by this disease. A total of 55 Streptomyces strains were isolated from potato scab lesions, among which 40 strains were pathogenic on progeny tubers. Among the pathogenic strain, 21 strains were identified as previously described S. scabies, 7 Strains as S. turgidiscabies, and 5 Strains as S. acidiscabies, while 7 strains were observed as having distinct phenotypic properties. These strains were classified into six distinct clusters based on phenotypic characteristics and selected representative strains for each cluster. S. scabies (S33) had grey spores in a spiral chain. Mean-while, S. turgidiscabies (S27) had grey spores, S. acidiscabies (S71) had white spores, S. luridiscabiei (S63) had yellow-white spores, S. puniciscabiei (S77) had purple-red spores, and S. niveiscabiei (S78) had thin and compact white spores, all in a rectiflexuous chain. Pathogenicity was determined by the production of thaxtomin A and homologs of necl and ORFtnp genes. In TLC, representative strains S27, S71, S63, S77, and S78 produced a yellow band that co-migrated with the authentic thaxtomin A. However, thaxtomin A was not detected in chloroform extracts from oatmeal broth culture and Slice tuber tissue of S. luridiscabiei (S63) and S. puniciscabiei (S77) by HPLC analysis. In addition, no homologs of necl and ORFtnp genes in S. acidiscabies (S71), S. luridiscabiei (S63), S. puniciscabiei (S77), and S. niveiscabiei (S78) were detected by PCR and Southern hybridization analysis.

Isolation and Characterization of Starch-hydrolyzing Pseudoalteromonas sp. A-3 from the Coastal Sea Water of Daecheon, Republic of Korea (대한민국 대천 해안에서 분리한 전분 분해능을 갖는 Pseudoalteromonas sp. A-3 균주의 특징 및 동정)

  • Chi, Won-Jae;Park, Da-Yeon;Jeong, Sung-Cheol;Chang, Yong-Keun;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.317-323
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    • 2011
  • Strain A-3, an amylase-producing bacteria, was isolated from coastal seawater near Daecheon in the Republic of Korea. It was seen to possess a single polar flagella and grow well, on ASW-YP agar plates, at temperatures of between $20-37^{\circ}C$. However, it grew more slowly at the temperatures of $15^{\circ}C$ and $40^{\circ}C$. Similarly, it was observed to grow abundantly, in an Artificial Sea Water-Yeast extract-Peptone (ASW-YP) liquid medium, in a pH range of 6-9, but not grow at pHs of 4-5 and a pH of 10. Strain A-3 was noted as being close to Pseudoalteromonas phenolica O-$BC30^T$, Pseudoalteromonas luteoviolacea $NCIMB1893^T$, Pseudoalteromonas rubra $ATCC29570^T$, and Pseudoalteromonas byunsanensis $FR1199^T$, with 98.30%, 97.86%, 97.78%, and 97.25% similarities respectively, in its 16S rRNA sequence. A phylogenetic tree revealed that strain A-3 and P. phenolica O-$BC30^T$ belong to a clade. However, strain A-3 differed from P. phenolica O-$BC30^T$ in relation to a number of physiological characteristics. Strain A-3 exhibited no growth above 5% NaCl concentrations, no utilization of D-glucose, D-mannose, D-maltose, or D-melibose, and no lipase (C-14) activity. All of these properties strongly indicate that strain A-3 is distant from P. phenolica O-$BC30^T$ and thus led us to name it Pseudoalteromonas sp. A-3. Pseudoalteromonas sp. A-3 produces ${\alpha}$-amylase throughout growth. Maximal amylase activities of 144.48 U/mL and 149.20 U/mL were seen at pH 7.0 and $37^{\circ}C$, respectively. Pseudoalteromonas sp. A-3's high, stable production of ${\alpha}$-amylase in addition to its biochemical features, such as alkalitolerance, suggest that it is a good candidate for industrial applications.

Physiological Characteristics of Starter Isolated from Kimchi and Fermentation of Tofu with Isolated Starter (발효두부 제조용 Starter의 선발과 이를 이용한 두부의 발효특성)

  • Kang, Kyoung Myoung;Lee, Shin Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.11
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    • pp.1626-1631
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    • 2012
  • Sixty strains of lactic acid bacteria were isolated from kimchi and used as a starter for fermented tofu. Among the isolated strains, strain KL-6 showed antimicrobial activity against various pathogens, antioxidative activity, and viability in artificial gastric juice and artificial bile acid. The selected strain KL-6 was identified as Pediococcus acidilactici KL-6 by morphological and physiological tests, including Gram staining, catalase test, and 16S rRNA sequencing. The fermentation characteristics of tofu with a kimchi ingredient mixture (Control) consisting of red pepper, garlic, ginger, sugar, salt, jeotgal, and juice of chinese cabbage were compared with those of tofus inoculated with strain KL-6 and the kimchi ingredient mixture (TL) or a pre-fermented kimchi ingredient mixture (TPL) for 24 hr at $37^{\circ}C$. The pH levels of all tested tofu samples decreased after 1 week of fermentation, reaching 3.96 (control), 3.97 (TL), and 4.03 log cfu/g (TPL) after fermentation for 14 weeks at $20^{\circ}C$. Total aerobe content of fermented tofu increased until 2 weeks of fermentation, but decreased steadily thereafter. The number of lactic acid bacteria reached $10^6$ cfu/g after 1 week of fermentation in TL and TPL, whereas it took 2 weeks for the control. The number of lactic acid bacteria in all tested tofu samples reached $10^3$ cfu/g after 14 weeks of fermentation at $20^{\circ}C$. Coliform bacteria were not detected in TL or TPL after 1 week of fermentation. The sensory scores of TL and TPL were higher than that of control in terms of taste, flavor, texture, and overall acceptability. The sensory quality of TPL was the best among all tested fermented tofu samples.

Isolation of Photosynthetic Bacterium, Rhodopseudomonas palustris JK-1 and Researches on IAA and Carotenoid Production (광합성세균 Rhodopseudomonas palustis 분리 및 IAA와 Carotenoid 생성에 관한 연구)

  • Kim, Yu-Kyoung;Cho, Young-Yun;Kang, Ho-Jun;Kim, Jung-Sun;Yang, Sung-Nyun;Jwa, Chang-sook
    • Korean Journal of Organic Agriculture
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    • v.25 no.4
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    • pp.843-859
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    • 2017
  • The JK-1 isolate which was the best producer of indole-3-acetic acid and carotenoid among the 388 strains isolated from 28 wetlands in Jeju, was identified to be Rhodopseudomonas palustirs belongs to a typical group of non sulfur purple bacteria based on 16S sRNA sequencing. This study investigated the effect of different cultural conditions of pH, temperature, agitation, light and aeration on growth, IAA and carotenoid production of photosynthetic bacterium JK-1 for optimization of IAA and carotenoid production. It was found that growth, IAA, carotenoid, and bacteriochlorophyll production with light (3,000~3,500 Lux) and agitation (100 rpm) showed better results than those with dark/static or dark/agitation (100 rpm) in anaerobic conditions. The optimal pH, temperature and agitation speed for cell growth were 7, $30^{\circ}C$, 150 rpm, for IAA production were 9, $30^{\circ}C$, 150rpm and for carotenoid production were 6, $25^{\circ}C$, 50 rpm, cultured for 72 h under anaerobic light, respectively. The growth and IAA production were high in aerobic culture compared with anaerocic culture, whereas carotenoid and bacteriochlorophyll content were decreased extremely in aerobic condition (0.5~1 vvm). Subsequently, the optimal culture conditions for JK-1 were selected with pH 7, $30^{\circ}C$ and 100 rpm under anaerobic light and the effect on plant growth was tested by pot assay. Inoculation of JK-1 with 3% (v/v) level caused increase in shoot and root dry weigh that varied from 20%~58% to 40%~28% in young radish in camparison to uninoculated treatment at 50 days of growth. The study suggests that the JK-1 isolate may serve as efficient biofertilizer inoculants to promote plant growth.

Isolation and Characterization of Bacillus Strain as a Potential Biocontrol Agent (환경친화적 미생물농약으로서의 잠재성을 가진 세균의 분리 및 특성)

  • Lee, Ye-Ram;Lee, Sang-Mee;Jang, Eun-Young;Hong, Chang-Oh;Kim, Keun-Ki;Park, Hyean-Cheal;g Lee, Sang-Mon;Kim, Young-Gyun;Son, Hong-Joo
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1408-1414
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    • 2015
  • In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.

Multiplex PCR of Endotracheal Aspirate for the Detection of Pathogens in Ventilator Associated Pneumonia (기계환기폐렴의 원인균 진단에서 인공기도 흡인액을 이용한 Multiplex PCR과 세균배양 결과의 비교)

  • Song, Ju Han;Myung, Soon Chul;Choi, Song Ho;Jeon, Eun Ju;Kang, Hyung Gu;Lee, Hye Min;Cho, Sung Keun;Choi, Jae Chol;Shin, Jong Wook;Park, In Won;Choi, Byoung Whui;Kim, Jae Yeol
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.3
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    • pp.194-199
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    • 2008
  • Background: Early identification of pathogens can improve the prognosis of patients with ventilator associated pneumonia (VAP). In the present study, we evaluated the feasibility of performing multiplex PCR for endotracheal aspirates to detect three important pathogens (P. aeruginosa, K. pneumoniae and MRSA) in patients with VAP. Methods: The endotracheal aspirates of 24 patients were collected within 24 hours of the diagnosis of VAP for performing multiplex PCR. Forward and reverse primers were designed to target the specific site of each pathogen (the oprL gene for P. aeruginosa, 16S rRNA for K. pneumoniae and the mec gene for MRSA). We analyzed the clinical data of the VAP patients, including the culture reports for the endotracheal aspirates. Results: Twenty-four patients (M:F=18:6, mean age=$70{\pm}11$) with VAP were enrolled. Pathogens were isolated from 11 patients (P. aeruginosa in 2, K. pneumoniae in 1, MRSA in 2, other enteric Gram negative bacilli in 3, S. pneumoniae in 2 and mixed infection in 1). Multiplex PCR detected three cases of P.aeruginosa (2 cases coincided with the culture reports) and four cases of K. pneumoniae (1 matched with the culture report). PCR detected two MRSA cases, which did not coincide with the culture reports. Conclusion: Multiplex PCR of the endotracheal aspirate showed some ability to detect Gram negative bacilli, although caution is required when interpreting the results.