• Title/Summary/Keyword: 16S ribosomal DNA

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Production of Violacein by a Novel Bacterium, Massilia sp. EP15224 Strain (Violacein을 생산하는 Massilia sp. EP15224 균주)

  • Yoon, Sang-Hong;Baek, Hee-Jin;Kwon, Soon-Wu;Lee, Chang-Muk;Sim, Joon-Soo;Hahn, Bum-Soo;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.317-323
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    • 2014
  • Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

Isolation of Lactic Acid Bacteria Showing Antioxidative and Probiotic Activities from Kimchi and Infant Feces

  • Ji, Keunho;Jang, Na Young;Kim, Young Tae
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1568-1577
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    • 2015
  • The purpose of this study was to investigate lactic acid bacteria with antioxidative and probiotic activities isolated from Korean healthy infant feces and kimchi. Isolates A1, A2, S1, S2, and S3 were assigned to Lactobacillus sp. and isolates A3, A4, E1, E2, E3, and E4 were assigned to Leuconostoc sp. on the basis of their physiological properties and 16S ribosomal DNA sequence analysis. Most strains were confirmed as safe bioresources through nonhemolytic activities and non-production of harmful enzymes such as β-glucosidase, β-glucuronidase and tryptophanase. The 11 isolates showed different resistance to acid and bile acids. In addition, they exhibited antibacterial activity against foodborne bacteria, especially Bacillus cereus, Listeria monocytogenes, and Escherichia coli. Furthermore, all strains showed significantly high levels of hydrophobicity. The antioxidant effects of culture filtrates of the 11 strains included 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging capacity, 2.2'-azino-bis (2-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation scavenging activity, and superoxide dismutase activity. The results revealed that most of the culture filtrates have effective scavenging activity for DPPH and ABTS radicals. All strains appeared to have effective superoxide dismutase activity. In conclusion, the isolated strains A1, A3, S1, and S3 have significant probiotic activities applicable to the development of functional foods and health-related products. These strains might also contribute to preventing and controlling several diseases associated with oxidative stress, when used as probiotics.

Analysis of ITS DNA Sequences of Korean Oxalis Species (Oxalidaceae) (한국산 괭이밥속(Oxalis) 식물 ITS DNA 염기서열 분석)

  • Koo, Jachoon;Chae, Mi Suk;Lee, Jeoung-Ki;Whang, Sung Soo
    • Korean Journal of Plant Taxonomy
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    • v.37 no.4
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    • pp.419-430
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    • 2007
  • This study was conducted to know the taxonomic features of nuclear ribosomal ITS DNA sequences, ITS1, ITS3 and 5.8S regions, as to nine individuals belonging to four Oxalis species in Korea and an induced species. Sequences of the same regions of sixteen taxa deposited in GenBank were also aligned with those of Korean species as outgroups. The length of ITS sequences aligned in this study is 679 by in total. Evidences, from not only the sequence similarities and divergences but also the phylogenetic and statistical treatments with ITS sequences aligned, were useful for the taxonomy of the genus. The similarity of sequences, among both cauline and acauline taxa, is high as 89% and 95% respectively, but between cauline and acauline taxa, relatively low in the range of 64~69%. The sequence divergences, among both cauline and acauline taxa, is also high as much as 0.36~0.42, but between both cauline and both acauline taxa, low as 0.04~0.06. Two groups between cauline and acauline taxa are paraphyletic, and each group makes a single Glade with a high bootstrap value. The analysis of variance, using ITS sequence aligned, revealed that taxa are significantly different in the level of 0.5%, and O. corymbosa, an induced speices, is also separated from the Korean taxa in the Duncan analysis.

Isolation and Identification of Three Pseudomonas koreensis Strains with Anti-microbial Activities Producing Inducers of the Expression of Egr-1 Gene (Egr-1 유전자의 발현 유도물질을 생산하는 항균성 저 영양 세균의 분리 및 동정)

  • Yoon, Sang-Hong;Kim, Dong-Gwan;Lee, Young-Han;Shin, Soon-Young;Kwon, Soon-Woo;Lee, Chang-Muk;Kang, Han-Chul;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.119-125
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    • 2011
  • The Egr-1 gene is known to be a transcription factor for activating the expression of many tumor-repressing genes. In this study, three strains activating the promoter of the Egr-1 gene were selected, through the use of Egr-1 luciferase reporter assay and western blotting, from amongst approximately 3,800 oligotrophic bacteria isolated from the cultivated soils of various regions within Korea. These strains were identified as Pseudomonas koreensis on the basis of phylogenetic tree analysis of their 16S ribosomal DNA sequences and biochemical characteristics analyses using a variety of commercial kits (API 20NE, ID 32GN, API ZYM kits). In addition, we discovered that these strains produced anti-bacterial activity against Bacillus subtilis, Staphylococcus aureus and Listeria monocytogenes.

The Complete Mitochondrial Genome and Molecular Phylogeny of the Flathead Platycephalus cultellatus Richardson, 1846 from Vietnam (Teleostei; Scorpaeniformes) (베트남 Platycephalus cultellatus Richardson, 1846 (Teleostei; Scorpaeniformes)의 전장 미토콘드리아 유전체와 분자계통)

  • Tran, Biet Thanh;Nguyen, Tu Van;Choi, Youn Hee;Kim, Keun-Yong;Heo, Jung Soo;Kim, Keun-Sik;Ryu, Jung-Hwa;Kim, Kyeong Mi;Yoon, Moongeun
    • Korean Journal of Ichthyology
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    • v.33 no.4
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    • pp.217-225
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    • 2021
  • The family Platycephalidae is a taxonomic group of economically important demersal flathead fishes that predominantly occupy tropical or temperate estuaries and coastal environments of the Indo-Pacific oceans and the Mediterranean Sea. In this study, we for the first time analyzed the complete mitochondrial genome (mitogenome) of the flathead Platycephalus cultellatus Richardson, 1846 from Vietnam by Next Generation Sequencing method. Its mitogenome was 16,641 bp in total length, comprising 13 protein-coding genes (PCGs), two ribosomal RNA genes, and 22 transfer RNA genes. The gene composition and order of the mitogenome were identical to those of typical vertebrates. The phylogenetic trees were reconstructed based on the concatenated nucleotide sequence matrix of 13 PCGs and the partial sequence of a DNA barcoding marker, cox1 in order to determine its molecular phylogenetic position among the order Scorpaeniformes. The phylogenetic result revealed that P. cultellatus formed a monophyletic group with species belonging to the same family and consistently clustered with one nominal species, P. indicus, and two Platycephalus sp. specimens. Besides, the cox1 tree confirmed the taxonomic validity of our specimen by forming a monophyletic clade with its conspecific specimens. The mitogenome of P. cultellatus analyzed in this study will contribute valuable information for further study on taxonomy and phylogeny of flatheads.

Morphological and molecular evidence of the hybrid origin of Crepidiastrum ×muratagenii in Korea (홍도고들빼기의 형태 다양성 및 잡종 기원의 분자 증거)

  • JANG, Young-Jong;PARK, Boem Kyun;SON, Dong Chan;CHOI, Byoung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.52 no.2
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    • pp.85-96
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    • 2022
  • The plant "Hong-do-go-deul-ppae-gi" has been considered as Crepidiastrum × muratagenii, a hybrid between C. denticulatum and C. lanceolatum, based on its morphological traits and geographical distribution. To reveal the hybrid origin of Hong-do-go-deul-ppae-gi, we examined additional morphological traits of this plant and its putative parents (C. denticulatum, C. lanceolatum, C. platyphyllum) and analyzed one nuclear ribosomal internal transcribed spacer (ITS) region and four chloroplast regions (trnT-L, trnL-F, rpl16 intron, and rps16 intron). As a result of examining the morphological traits, putative hybrid individuals were classified into three types based on the habit, cauline leaf, outer phyllary, and achene beak traits. A molecular analysis found that the ITS sequences of Type 1 and Type 2 individuals showed additive species-specific sites of C. denticulatum and C. lanceolatum. Plastid sequences of Type 1 and Type 2 individuals showed C. denticulatum and C. lanceolatum sequences, respectively. However, Type 3 individuals had ITS and plastid sequences corresponding to C. denticulatum. Accordingly, Type 1 and Type 2 individuals not only share morphological traits with C. denticulatum and C. lanceolatum but also show additive species-specific sites for C. denticulatum and C. lanceolatum, and not C. platyphyllum, supporting its origin as a hybrid between C. denticulatum and C. lanceolatum. Type 3 had morphological traits similar to other hybrid types but was distinguished with respect to outer phyllaries and demonstrated some resemblance to C. denticulatum. In a molecular analysis, Type 3 was found to be identical with regard to the sequence of C. denticulatum and was judged to be an ecological variation of C. denticulatum.

Purification of \alpha-Cyclodextrin Glucanotransferase Excreted from Themophilic Geobacillus thermosac-chalytycus and Characterization of Transglycosylation Reaction of Glucosides. (호열성 Geobacillus thermosacchalytycus가 생산하는 \alpha-Cyclodextrin Glucanotransferase의 분리정제와 당전이 반응 특성)

  • 이미숙;신현동;김태권;이용현
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.29-36
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    • 2004
  • $\alpha$-Cyclodextrin glucanotransferase excreted from a newly isolated Geobacillus thermosacchalytycus was purified through the ultrafiltraion, hydrophobic Sepharose CD-4B affinity chromatography, and gel filtration on Sephadex G-75, respectively. The molecular weight of the purified CGTase was 69 kDa and its N-terminal amino acid sequence was determined to be Asn-Leu-Asn-Lys-Val-Asn-Phe-Val-Ser-Asp-Val-Val-Val-Gln-Ile. The optimum pH and temperature were pH 6.0 and$ 60^{\circ}C$, respectively, and stably at the pH range of 6.0-8.0 and $60^{\circ}C$ in the presence of $Ca^{++}$. The excreted CGTase from the thermophilic G. thermosacchalytycus was $\alpha$-type showing a high coupling activity for the transglycosylation on various glucosides. The coupling reaction was carried out according to the random ternary complex mechanism.m.

Isolation and Identification of a Streptomyces sp. that Produces Antibiotics Against Multidrug - Resistant Acinetobacter baumannii (다제내성 Acinetobacter baumannii의 생장을 억제하는 항생물질을 생산하는 방선균의 분리.동정 및 항균효과)

  • Rhee, Ki-Hyeong
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.37-42
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    • 2011
  • I isolated the actinomycete strain KH223 from soil samples collected from the Kye Ryong mountain area. This strain is antagonistic to the multidrug-resistant Acinetobacter baumannii. KH223 was confirmed as belonging to the genus Streptomyces based on the scanning electronmicroscopy(SEM) observations of the diaminopimelicacid(DAP) type and morphological and physiological characteristics. Comparison of the 16S rDNA nucleotide sequences revealed that KH223 has a relationship with Streptomyces galbus. Production of antibiotics by KH223 was most favorable when cultured on a glucose, polypeptone, and yeast extract(PY) medium for 6 days at 27$^{\circ}C$. The supernatant was found to exhibit an antimicrobial effect on various kinds of bacteria and fungi. Particularly, butanol and ethylacetate extracts of KH223 and cyclo(trp-trp) exhibited significant activity against A. baumannii at concentration ranges of 0.8-12.5 ${\mu}g$/mL, 5.0-25 ${\mu}g$/mL and 12.5${\rightarrow}$100 ${\mu}g$/mL, respectively. Moreover, in contrast to cyclo(trp-trp) had shown to activity against Micrococcus luteus JCM 1464 at the concentration of 12.5 ${\mu}g$/mL, the butanol extract of KH223 showed significant activity against Bacillus subtilis IAM 1069 and Micrococcus luteus JCM 1464 at the concentration of 0.4 and 0.8 ${\mu}g$/mL, respectively. These results suggest that KH223 may have a great potential in the production of new antibiotics to combat multidrug-resistant pathogens and further studies may be warranted for the same.

Probiotic Potential of Lactobacillus Isolates (Lactobacillus 분리균주의 프로바이오틱스로서의 가능성 검토)

  • Bang, Ji-Hun;Shin, Hwa-Jin;Choi, Hye-Jung;Kim, Dong-Wan;Ahn, Cheol-Soo;Jeong, Young-Kee;Joo, Woo-Hong
    • Journal of Life Science
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    • v.22 no.2
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    • pp.251-258
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    • 2012
  • The purpose of this study was to investigate the probiotic properties of lactic acid bacterial strains isolated from animal feces. BCNU 9041 and BCNU 9042 isolates were assigned to Lactobacillus brevis on the basis of their physiological properties and 16S ribosomal DNA sequences analysis. They were confirmed as safe bioresources because of their non-hemolytic activities and non-production of harmful ${\beta}$-glucosidase, ${\beta}$-glucuronidase, tryptophanase, or urease. These isolates were also highly resistant to acid (at pH 2.5) and bile acids (at concentration of 0.3%, 0.6%, and 1% oxgall). In addition, they exhibited good antibacterial activity against food-borne bacteria, especially Bacillus cereus, Listeria monocytogenes, and Shigella sonnei. Furthermore, it was demonstrated that they have the highest levels of hydrophobicity and that they showed bile salt hydrolytic and cholesterol assimilation activity. These results suggest that BCNU 9041 and 9042 have good potential for application in functional foods and health-related products.