• 제목/요약/키워드: 16S rDNA sequences

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전통 장류로부터 Exopolysaccharide 생성 유산균의 분리 및 특성 (Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Korean Soy Sauce and Soybean Paste)

  • 윤혜주;이유정;여수환;박혜영;박희동;백성열
    • 한국미생물·생명공학회지
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    • 제41권2호
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    • pp.190-197
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    • 2013
  • 전통 장류에서 점성물질을 분비하는 유산균을 분리하고 높은 활성을 보이는 N45-10, K6-7 및 N58-5의 3균주를 선발하였다. 선발한 3균주의 16S rDNA의 염기서열을 분석한 결과 N45-10은 Leuc. citreum, K6-7, N58-5는 Leuc. mesenteroides로 동정되었다. 산 저항성과 인공위액 저항성은 3균주 중에서 Leuc. citreum N45-10이 높은 생균수($10^5-10^6$ CFU/ml)를 나타내어 산 저항성과 인공위액 저항성이 가장 우수하게 나타났으며, 인공 담즙 저항성은 Leuc. citreum N45-10은 높은 생균수($10^3-10^4$ CFU/ml)를 유지하여 담즙액 저항성이 우수하게 나타났다. 3균주들 중에서 Leuc. citreum N45-10은 EPS 생성량이 가장 많았고, MRS 배지에서 배양하여 EPS가 생성되지 않았을 때보다 슈크로오스 배지에서 배양하여 EPS를 생성하였을 때 생균수가 더 높게 나타났다. 이것은 EPS 생성이 유산균의 세포벽 주위에 보호막으로 작용한 결과로 생각된다. 선발 유산균 3주의 EPS 생산량은 슈크로오스 액체배지에서 각각 16.173, 8.167, 3.652 g/L였으며, Leuc. citreum N45-10은 글루코오스로만 이루어진 homo-polysaccharide, Leuc. mesenteroides K6-7과 N58-5는 프락토오스, 글루코오스로 구성된 hetero-polysaccharides로 동정되었다. 선발 유산균 3주 모두는 용혈 음성반응을 나타내고, 젤라틴 액화능을 나타내지 않아 안전성이 확인되었다.

Agar를 분해하는 swarming 박테리아 균주의 특성과 동정 (Characterization and Identification of an Agar-Degrading Motile Bacteria Strain)

  • 강성완;유아영;유종언;강호영
    • 생명과학회지
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    • 제22권2호
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    • pp.259-265
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    • 2012
  • 환경에서 분리된 CK214 균주는 1.5% (w/v) agar가 포함되어 있는 LB 평판배지에서 빠르게 이동하는 특징을 가지며, agar 고체평판배지 위의 CK214 균주의 집락 주위로 움푹한 투명환이 관찰되었다. 이 균주는 단일 탄소원으로 agar만이 첨가된 최소 배지에서 잘 자랐으며, DNS 법을 이용하여 CK214 균주의 외부추출성분이 agar 분해활성을 가진다는 것을 확인하였다. CK214 균주는 다양한 농도의 agar (0.5, 1.0, 1.5 2.0% w/v)가 포함된 고체평판 배지에서 swarming 운동을 하였다. CK214 균주를 동정하기 위해 그람염색과 현미경 관찰, 생화학적 분석(API), 16S rRNA 염기서열분석에 기초한 계통발생학적 분석을 수행하였다. 이를 통해 CK214 균주는 그람 양성의 간균으로, Paenibacillus 속에 포함되었으며 Paenibacillus lactis MB 2035와 가장 가까운 연관성을 보이는 것을 확인할 수 있었다. 또한 CK214 균주는 agar 고체표면에서 주모성의 편모를 형성하는 것을 투과 전자 현미경(TEM)을 통해 관찰하였다. CK214 균주의 agarase 활성과 운동성의 연관성에 관한 앞으로의 연구를 위해 transposon random mutagenesis에 의한 agar 분해활성 결손 돌연변이주를 구축하였다.

한국 여수에서 채집된 매가오리과 (Myliobatidae) 어류 첫기록종, Mobula thurstoni (First Record of the Smoothtail Mobula, Mobula thurstoni (Myliobatiformes: Myliobatidae) in Southern Korea)

  • 명세훈;송영선;강충배;최홍인;김종관;윤문근;임재복;한동진
    • 한국어류학회지
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    • 제33권2호
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    • pp.148-154
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    • 2021
  • 매가오리목 매가오리과에 속하는 Mobula thurstoni 2개체(1770~1850 mm 체반폭)가 2018년 9월 전라남도 여수시 연도 연안에서 정치망으로 채집되었다. 이 종은 가슴지느러미의 앞부분이 이중 굴곡이고, 등지느러미 바로 뒤 꼬리 시작부분에 가시가 없으며, 등지느러미 끝부분에는 흰색이고, 그리고 등쪽의 체색이 어두운 남색을 띤다. Mobula kuhlii와 가장 형태적으로 유사하였지만, 가슴지느러미 앞부분에 이중 굴곡을 가지고 있다는 점(vs. 직선이거나 약간의 굴곡을 가진다)과 등쪽 체색이 어두운 남색을 띤다는 점(vs. 회갈색)에서 잘 구분된다. 또한, 이 종은 M. kuhlii와 미토콘드리아 16S rRNA 영역에서 유전적 거리 0.030~0.069의 차이를 보여 구분되었다. 이 종의 새로운 국명으로 '매끈꼬리쥐가오리'를 제안한다.

Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

  • Chung, Eu Jin;Hossain, Mohammad Tofajjal;Khan, Ajmal;Kim, Kyung Hyun;Jeon, Che Ok;Chung, Young Ryun
    • The Plant Pathology Journal
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    • 제31권2호
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    • pp.152-164
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    • 2015
  • Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.

한국인 신생아의 분변 미생물 균총 (Fecal Microflora of Korean Neonates)

  • 이승규;정석근;오미화;김동훈;강대경;이완규;함준상
    • Journal of Dairy Science and Biotechnology
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    • 제27권2호
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    • pp.1-6
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    • 2009
  • Probiotic bacteria have been administered to neonates to serve as maturational stimuli for the developing gut and intestinal immune system, establish and develop the intestinal microbiota, and mediate host-microbe interactions; further, these bacteria have shown beneficial effects In the treatment and reduction of the risk of infectious diseases, necrotizing enterocolitis, and atopic disease. An LAB isolation project to identify effective lactic acid bacteria for Korean people is in progress. The average total counts of lactic acid bacteria, lactobacilli, bifidobacteria, and coliforms in the fecal samples from 2 provinces were estimated as 8.31, 5.98, 8.13, and 3.01 CFU/g. Additional samples from other provinces will be analyzed to examine the changes in the lactic bacterial counts according to the area, sex of the neonate, mode of delivery, and type of feeding. A database containing the 16S rDNA sequences and the ribosomal protein profile of all the lactic acid bacteria isolated from fecal samples will be constructed. For the effective use of probiotics, a number of clinical studies are needed to formulate guidelines for strain, subject, purpose, and dose.

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Isolation of a Nisin-Producing Lactococcus lactis Strain from Kimchi and Characterization of its nisZ Gene

  • Lee, Kwang-Hee;Moon, Gi-Seong;An, Jong-Yun;Lee, Hyong-Joo;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Jong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제12권3호
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    • pp.389-397
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    • 2002
  • Bacteriocin-producing lactic acid bacteria were isolated from kimchi. One isolate producing the most efficient bacteriocin was identified and named Lactococcus lactis B2, based on the biochemical properties and 16S rDNA sequences. The B2 bacteriocin inhibited many different Gram positive bacteria including Lactococcus, Lactobacillus, Leuconostoc, Enterococcus, Streptococcus, and Staphylococcus, but did not inhibit Gram-negative bacteria. The bacteriocin was maximally produced at temperatures between $25^{\circ}C\;and\;30^{\circ}C$ and at the initial pH of 7.0. Ninety $\%$ of the activity remained after 10 min of heat treatment at $121^{\circ}C,\;and\;100\%$, after 1 h exposure to organic solvents. The bacteriocin was purified from culture supernatant by ammonium sulfate precipitation, CM Sepharose column chromatography, ultrafiltration, and finally, by reverse-phase HPLC. A 1.58-kb fragment was amplified from B2 chromosome by using a primer set designed from the published nisA sequence. Sequencing result showed that the fragment contained the whole nisZ and 5' portion of nisB, whose gene product was involved in postmodification of nisin. The upstream sequence, however, was completely different from those of reported nisin genes.

Genetic and Phenotypic Diversity of Parathion-Degrading Bacteria Isolated from Rice Paddy Soils

  • Choi, Min-Kyeong;Kim, Kyung-Duk;Ahn, Kyong-Mok;Shin, Dong-Hyun;Hwang, Jae-Hong;Seong, Chi-Nam;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • 제19권12호
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    • pp.1679-1687
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    • 2009
  • Three parathion-degrading bacteria and eight pairs of bacteria showing syntrophic metabolism of parathion were isolated from rice field soils, and their genetic and phenotypic characteristics were investigated. The three isolates and eight syntrophic pairs were able to utilize parathion as a sole source of carbon and energy, producing p-nitrophenol as the intermediate metabolite during the complete degradation of parathion. Analysis of the 16S rRNA gene sequence indicated that the isolates were related to members of the genera Burkholderia, Arthrobacter, Pseudomonas, Variovorax, and Ensifer. The chromosomal DNA patterns of the isolates obtained by polymerasechain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences were distinct from one another. Ten of the isolates had plasmids. All of the isolates and syntrophic pairs were able to degrade parathion-related compounds such as EPN, p-nitrophenol, fenitrothion, and methyl parathion. When analyzed with PCR amplification and dot-blotting hybridization using various primers targeted for the organophosphorus pesticide hydrolase genes of previously reported isolates, most of the isolates did not show positive signals, suggesting that their parathion hydrolase genes had no significant sequence homology with those of the previously reported organosphophate pesticide-degrading isolates.

Genetic Diversity of Cultivable Plant Growth-Promoting Rhizobacteria in Korea

  • Kim, Won-Il;Cho, Won-Kyong;Kim, Su-Nam;Chu, Hyo-Sub;Ryu, Kyoung-Yul;Yun, Jong-Chul;Park, Chang-Seuk
    • Journal of Microbiology and Biotechnology
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    • 제21권8호
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    • pp.777-790
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    • 2011
  • To elucidate the biodiversity of plant growth-promoting rhizobacteria (PGPR) in Korea, 7,638 bacteria isolated from the rhizosphere of plant species growing in many different regions were screened. A large number of PGPR were identified by testing the ability of each isolate to promote the growth of cucumber seedlings. After redundant rhizobacteria were removed via amplified rDNA restriction analysis, 90 strains were finally selected as PGPR. On the basis of 16S ribosomal RNA sequences, 68 Gram-positive (76%) and 22 Gram-negative (24%) isolates were assigned to 21 genera and 47 species. Of these genera, Bacillus (32 species) made up the largest complement, followed by Paenibacillus (19) and Pseudomonas (11). Phylogenetic analysis showed that most of the Grampositive PGPR fell into two categories: low- and high- G+C (Actinobacteria) strains. The Gram-negative PGPR were distributed in three categories: ${\alpha}$-proteobacteria, ${\beta}$- proteobacteria, and ${\gamma}$-proteobacteria. To our knowledge, this is the largest screening study designed to isolate diverse PGPR. The enlarged understanding of PGPR genetic diversity provided herein will expand the knowledge base regarding beneficial plant-microbe interactions. The outcome of this research may have a practical effect on crop production methodologies.

논에서 분리한 메탄산화세균 Methylomonas sp. SM4의 특성과 메탄올 생합성 (Characterization and Methanol Biosynthesis of a Methane-Oxidizing Bacterium, Methylomonas sp. SM4, Isolated from Rice Paddy Field Soil)

  • 박성민;;김시욱
    • KSBB Journal
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    • 제32권2호
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    • pp.124-132
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    • 2017
  • A methane-oxidizing bacterium was isolated from rice paddy field soil around Jeollanam-do province, Korea, and characterized. The isolate was gram-negative, orange pigmented and short rod ($1.1-1.2{\times}1.6-1.9{\mu}m$). It was catalase and urease-negative but oxidase-positive. The strain utilized methane and methanol as sole carbon and energy sources. It had an ability to grow with an optimum pH 7.0 and an optimum growth temperature $30^{\circ}C$. The strain was resistant to antibiotic polymyxin B but sensitive to streptomycin, kanamycin, ampicillin, chloramphenicol and rifampicin. The isolate required copper for their growth with concentration range of $2-25{\mu}M$, with an optimum of $10{\mu}M$. Under optimal culture condition, specific cell growth rate and generation time were found to be $0.046hr^{-1}$ and 15.13 hr, respectively. Phylogenetic analysis based on 16S rDNA sequences indicated that the strain formed a tight phylogenetic lineage with Methylomonas koyamae with a value of 99.4% gene sequence homology. So, we named the isolate as Methylomonas sp. SM4. 8.6 mM methanol was accumulated in the reaction mixture containing 70 mM sodium formate and 40 mM $MgCl_2$ (MDH inhibitor) under atmosphere of methane:air (40:60) mixture for 24 hr at $30^{\circ}C$.

Genetic and Phenotypic Diversity of Carbofuran-Degrading Bacteria Isolated from Agricultural Soils

  • Shin, Dong-Hyeon;Kim, Dong-Uk;Seong, Chi-Nam;Song, Hong-Gyu;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • 제22권4호
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    • pp.448-456
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    • 2012
  • Thirty-seven carbofuran-degrading bacteria were isolated from agricultural soils, and their genetic and phenotypic characteristics were investigated. The isolates were able to utilize carbofuran as a sole source of carbon and energy. Analysis of the 16S rRNA gene sequence indicated that the isolates were related to members of the genera Rhodococcus, Sphingomonas, and Sphingobium, including new types of carbofuran-degrading bacteria, Bosea and Microbacterium. Among the 37 isolates, 15 different chromosomal DNA patterns were obtained by polymerase chain reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences. Five of the 15 representative isolates were able to degrade carbofuran phenol, fenoxycarb, and carbaryl, in addition to carbofuran. Ten of the 15 representative isolates had 1 to 8 plasmids. Among the 10 plasmid-containing isolates, plasmid-cured strains were obtained from 5 strains. The cured strains could not degrade carbofuran and other pesticides anymore, suggesting that the carbofuran degradative genes were on the plasmid DNAs in these strains. When analyzed with PCR amplification and dot-blot hybridization using the primers targeting for the previously reported carbofuran hydrolase gene (mcd), all of the isolates did not show any positive signals, suggesting that their carbofuran hydrolase genes had no significant sequence homology with the mcd gene.