• 제목/요약/키워드: 16s rRNA 유전자

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Characteristics and functional analysis of Bacillus strains from the fermented soybean products, Cheonggukjang (전통 발효 청국장으로부터 분리한 Bacillus 균주들의 특성 및 기능 분석)

  • Moon, Ji-Young;Kwon, Soon-Wo;Hong, Seung-Beom;Seok, Soon-Ja;Kim, Jeong-Seon;Kim, Soo-Jin
    • Korean Journal of Microbiology
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    • 제51권3호
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    • pp.300-307
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    • 2015
  • For selecting Bacillus strains producing high-quality Cheonggukjang, 8 strains were isolated from the different Cheonggukjang samples. Seven of them exhibited the highest 16S rRNA gene sequence similarity value of over 99.9% to Bacillus subtilis subsp. subtilis and one of them showed the similarity to B. licheniformis. All the strains showed positive activities for amylase, cellulase, protease and lipase, and 6 strains are positive for fibrinolytic activity. To confirm the safety of the strains isolated from the samples of Cheonggukjang which are manufactured by traditional method, strains were analyzed for the presence of seven toxin genes of Bacillus cereus and results were found negative. And 7 strains did not produce at all or merely produce both histamine and tyramine, the representative biogenic amines. Biogenic amine degradation analysis by HPLC revealed that, most of them exhibited tyramine degradation activity. For Cheonggukjang fermented by artificial inoculation of selected strains, fermentation property, sensory test, volatile basic nitrogen production and metabolic profiles by $^1H-NMR$ were tested. Seven strains were confirmed to make high-quality Cheonggukjang.

Antimicrobial and Biogenic Amine-Degrading Activity of Bacillus licheniformis SCK B11 Isolated from Traditionally Fermented Red Pepper Paste (전통고추장에서 유해균 억제 및 Biogenic Amines 분해 능력을 가지는 Bacillus licheniformis SCK B11의 분리)

  • Kim, Yong-Sang;Jeong, Jin-Oh;Cho, Sung-Ho;Jeong, Do-Yeon;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • 제48권2호
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    • pp.163-170
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    • 2012
  • In order to inhibit the growth of pathogens and degrade biogenic amines during the fermentation of soybean products, an isolate with antimicrobial activity against pathogens and biogenic amine-degrading property was obtained from 83 traditionally fermented soybean products. The morphological and biochemical tests and the phylogenetic relationship among 16S rRNA gene sequences indicated that the isolate named as the strain SCK B11 was most closely related to Bacillus licheniformis. The cell-free supernatant of two day cultures was active against several pathogens including Enterococcus faecalis, Listeria monocytosis, Micrococcus luteus, Pseudomonas aeruginosa, Bacillus cereus, and Staphylococcus aureus. PCR analysis was conducted to determine relatedness to antimicrobial lantibiotics and biosurfactants produced by Bacillus spp., but showed negative for the genes encoding surfactin, lichenysin, and lichenicidine. Electron microscopic observation indicated that the antimicrobial agent seemed to attack the membrane of the pathogens, leaving the ghost or shrunken cells. The strain was found to degrade histamine by 72% and tyramine by 66% in the cooked soybean containing 5.3% of biogenic amine over 10 days of fermentation time. The use of selected strain would be a potential control measure in manufacturing traditionally fermented soybean products that are difficult to control pathogens and biogenic amine levels.

Bacterial Spot Disease of Green Pumpkin by Pseudomonas syringae pv. syringae (Pseudomonas syringae pv. syringae에 의한 애호박 세균점무늬병)

  • Park, Kyoung-Soo;Kim, Young-Tak;Kim, Hye-Seong;Lee, Ji-Hye;Lee, Hyok-In;Cha, Jae-Soon
    • Research in Plant Disease
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    • 제22권3호
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    • pp.158-167
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    • 2016
  • A pathogen that causes a new disease on green pumpkin in the nursery and the field was characterized and identified. Symptoms of the disease on green pumpkin were water soaking lesions and spots with strong yellow halo on leaf, brown lesion on flower, and yellow spot on fruit. The bacterial isolates from the leaf spot were pathogenic on the 8 curcubitaceae crop plants, green pumpkin, figleaf gourd, wax gourd, young pumpkin, zucchini, cucumber, melon, and oriental melon, whereas they did not cause the disease on sweet pumpkin and watermelon. They were Gram-negative, rod shape with polar flagella, fluorescent on King's B agar and LOPAT group 1a by LOPAT test. Their Biolog substrate utilization patterns were similar to Pseudomonas syringae pv. syringae's in Biolog database. Phylogenetic trees with 16S rRNA gene sequences and multilocus sequence typing (MLST) with nucleotide sequences of 4 housekeeping genes, gapA, gltA, gyrB, rpoD and those of P. syringae complex strains in the Plant Associated and Environmental Microbes Database (PAMDB) showed that the green pumpkin isolates formed in the same clade with P. syringae pv. syringae strains. The clade in MLST tree was in the genomospecies 1 group. The phenotypic and genotypic characteristics suggested that the isolates from green pumpkin lesion were P. syringae pv. syringae.

Comparison of Bacterial Communities in Beach Sands along the East Coast of North Gyeongsang Province (경상북도 동해안 해변모래에 서식하는 미생물 군집 비교)

  • Khang, Yongho
    • Korean Journal of Microbiology
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    • 제50권4호
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    • pp.376-380
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    • 2014
  • Marine beach sands with bacterial pathogens may cause increased outcomes of illness among beachgoers in summer. In this study, pyrosequencing of 16S ribosomal DNAs extracted from 12 beach sands was performed to understand how the environmental factors of wastewaters or human wastes affected the distribution of bacterial communities at the beach of North Gyeongsang province (Yeongdeok and Pohang counties) in the middle of October. It was found that Acidobacteria were dominantly distributed in the sands near the clean seawaters, Proteobacteria in the sands near the river waters, Cyanobacteria in the sands near the wastewaters, and Bacteroidetes in the sands near the beach park. Other phyla groups such as Actinobacteria, Chlorobi, Deferribacteres, Deinococcus-thermus, Firmicutes, Gemmatimonadetes, Nitrospirae, and/or Verrucomicrobia were distributed at low relative abundance (1-5%).

Isolation and identification of β-glucosidase producing halophilic Roseivivax roseus (β-Glucosidase를 생성하는 호염성 Roseivivax roseus 균주의 분리 및 분류동정)

  • Cho, Geon-Yeong;Han, Song-Ih
    • Korean Journal of Microbiology
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    • 제51권2호
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    • pp.141-147
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    • 2015
  • Four halophilic bacteria were isolated from a salt water tank of more than 25% above salinity used for production of salt. HJS1 and HJS6 strains were identified as having ${\beta}$-glucosidase producing capabilities at high salinity. ${\beta}$-Glucosidase produced from these bacterial strains showed the best activity at 56-79 U/ml in NaCl (0-5%), showing the highest ${\beta}$-glucosidase activity at NaCl 3%. A salt tolerant ${\beta}$-glucosidase can maintain at least 75% activity of the enzyme in 0-20% NaCl concentration. The 16S rRNA gene sequences of strains HJS1 and HJS6 shows 99.8% similarity with Roseivivax roseus $BH87090^T$. Those sequences were registered as AB971835 and AB971836 in the NCBI GenBank. DNA-DNA hybridization test revealed that both strains showed 90.1 to 90.3% hybridization values with R. roseus $BH87090^T$, which was the closest phylogenetic neighbor. Major Cellular fatty acids of strains HJS1 and HJS6 were $C_{16:0}$, $C_{18:1}$ ${\omega}7c$, $C_{19:0}$ cyclo ${\omega}8c$ and 11-methyl $C_{18:1}$ and the major quinone was Q-10. Their fatty acid composition and quinone were very similar to Roseivivax roseus $BH87090^T$. Meanwhile, Roseivivax roseus $BH87090^T$ did not produce any ${\beta}$-glucosidase. Based on the molecular and chemotaxonomic properties, strains HJS1 and HJS6 were identified as members of Roseivivax roseus.

Evaluation of the Effect of Mine Drainage on the Aquatic Environment by Quantitative Real-time PCR (실시간 정량 중합효소연쇄반응을 이용한 광산 배수의 수계 영향 평가)

  • Han, Ji-Sun;Seo, Jang-Won;Ji, Won-Hyun;Park, Hyun-Sung;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • 제32권2호
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    • pp.121-130
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    • 2010
  • Metals and sulfate can be considerably dissolved at low pH condition in the acid mine drainage(AMD) and it would make an environmental problems. There are only few of acid mine drainage treatment systems in Korea which are operating, but these still have an effect on the surrounding stream. In this study, quantification of indicator microorganisms was conducted to judge the environmental impact of AMD on microflora by quantitative real-time PCR in the drainage samples of four mines and the water samples of each surrounding stream. Two species of iron reducing bacteria(Rhodoferax ferrireducens T118 and Acidiphilium cryptum JF-5) were selected for indicator bacteria based on 16S rRNA cloning analysis, and sulfate reducing bacteria(Desulfosporosinus orientus), iron and sulfur oxidizing bacteria(Acidothiobacillus ferrooxidans) and iron oxidizing bacteria(Leptosprillum ferrooxidans) were included into indicator since these were found in the previous studies on the mining area. Thereafter, the comparative analysis of four mines were established by the microbiological variation index and it was determined that the biological environment effect of AMD is highest in Samtan mine which doesn t contain treatment system by the value.

Identification of a new marine bacterium Ruegeria sp. 50C-3 isolated from seawater of Uljin in Korea and production of thermostable enzymes (대한민국 울진 연안 해양에서 분리한 해양 미생물 Ruegeria sp. 50C-3의 동정 및 내열성 효소 생산)

  • Chi, Won-Jae;Kim, Jong-Hee;Park, Jae-Seon;Hong, Soon-Kwang
    • Korean Journal of Microbiology
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    • 제52권3호
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    • pp.344-351
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    • 2016
  • A marine bacterium, designated as strain 50C-3, was isolated from a seawater sample collected from the East Sea of South Korea. The strain is a Gram-negative, aerobic, yellow colored polar-flagellated bacterium that grows at $20-50^{\circ}C$ and pH 5.5-8.5. Optimal growth occurred at $40-50^{\circ}C$, at pH 6.5-7.5, and in the presence of 2% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, the isolate was considered to represent a member of the genus Ruegeria. The result of this analysis showed that strain 50C-3 shared 99.4% and 96.98% sequence similarity with Ruegeria intermedia CC-GIMAT-$2^T$ and Ruegeria lacuscaerulensis ITI-$1157^T$, respectively. Furthermore, strain 50C-3 showed clear differences from related strains in terms of several characteristics such as motility, carbon utilization, enzyme production, etc. The DNA G+C content was 66.7 mol%. Chemotaxonomic analysis indicated ubiquinone-10 (Q-10) as the predominant respiratory quinone. Based on phenotypic, chemotaxonomic, and phylogenetic characteristics, the isolate represents a novel variant of the Ruegeria intermedia CC-GIMAT-$2^T$, for which we named Ruegeria sp. 50C-3 (KCTC23890=DSM25519). Strain 50C-3 did not produce cellulase and agarase, but produced alkaline phosphatase, ${\alpha}$-galactosidase, and ${\beta}$-galactosidase. The three enzymes showed stable activities even at $50^{\circ}C$ and thus regarded as thermostable enzymes. Especially, the ${\beta}$-galactosidase activity enhanced by 1.9 times at $50^{\circ}C$ than that at $37^{\circ}C$, which may be very useful for industrial application.

Properties of Antimicrobial Substances Produced by Bacillus amyloliquefaciens CJW15 and Bacillus amyloliquefaciens SSD8 (Bacillus amyloliquefaciens CJW15와 SSD8이 만드는 항균물질들의 특성)

  • Liu, Xiaoming;Shim, Jae Min;Yao, Zhuang;Lee, Jae Yong;Lee, Kang Wook;Kim, Hyun-Jin;Ham, Kyung-Sik;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • 제44권1호
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    • pp.9-18
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    • 2016
  • Two Bacillus strains, CJW15 and SSD8, with strong antibacterial activities were isolated from cheonggukjang. Both were identified as B. amyloliquefaciens strains after gene sequencing of rRNA and recA. CJW15 strongly inhibited the growth of B. cereus (ATCC14579), Listeria monocytogenes (ATCC19111), and Lactococcus lactis (ATCC11454). In comparison, SSD8 inhibited the growth of B. cereus (ATCC14579) and Enterococcus faecium (ATCC19953). The antibacterial activities of the two strains were not affected when exposed to a temperature of $100^{\circ}C$ for 15 min and were quite stable in acidic (pH 3) and alkaline (pH 12) pH conditions. Enzymatic treatments (trypsin, pepsin, proteinase K, and protease) had no effect on the activity of CJW15, but reduced the activity of SSD8 by half. Both isolates possess genes involved in the synthesis of lipopeptides (e.g. surfactin, fengycin, iturin, and iturin A), and genes encoding subtilin, a bacteriocin. Moreover, both isolates have fibrinolytic activities as well.

Production and Characterization of Mannanase from a Bacillus sp. YB-1401 Isolated from Fermented Soybean Paste (된장 분리균 Bacillus sp. YB-1401의 Mannanase 생산성과 효소특성)

  • Joen, Ho Jin;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • 제42권2호
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    • pp.99-105
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    • 2014
  • A Bacillus strain capable of hydrolyzing locust bean gum was isolated as a producer of extracellular mannanase by way of an enrichment culture in an acidic medium from homemade soybean pastes. The isolate YB-1401 showed a biochemical identity of 61.1% with Brevibacillus laterosporus, while the nucleotide sequence of its 16S rDNA had the highest similarity with that of Bacillus amyloliquefaciens. The mannanase productivity of the Bacillus sp. YB-1401 was drastically increased by mannans. Particularly, maximum mannanase productivity was reached at approximately 265 U/ml in LB medium supplemented with konjac glucomannan (4.0%). The mannanase was the most active at $55^{\circ}C$ and pH 5.5. Mannanase activity was completely maintained after pre-incubation at pH 3.5 to 11.0 for 1 h. The predominant products resulting from the mannanase hydrolysis were mannobiose and mannotriose for LBG, guar gum or mannooligosaccharides. A small amount of mannose was also detected in the hydrolyzates.