• Title/Summary/Keyword: l6S rDNA

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Isolation, Identification, and Characterization of Weissella Strains with High Ornithine Producing Capacity from Kimchi (김치로부터 오르니틴 생성능을 갖는 Weissella 속 균주의 분리, 동정 및 특성)

  • Yu, Jin-Ju;Park, Hyoung-Ju;Kim, Su-Gon;Oh, Suk-Heung
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.339-345
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    • 2009
  • Two lactic acid bacteria (LAB) with high ornithine-producing capacity were isolated from kimchi. Examination of the biochemical features using an API kit indicated that the strains belonged to the members of Weissella genus. They were gram positive, short rod-type bacteria, and able to grow anaerobically with $CO_2$ production. The isolates grew well on MRS broth at $25\sim37^{\circ}C$ and pH of 6.0~7.0. The optimum temperature and pH for growth are $30^{\circ}C$ and pH 6.5. The isolates fermented arabinose, ribose, xylose, glucose but not cellobiose, galactose, raffinose, or trehalsoe. The 16S rDNA sequences of isolates showed 99.6% and 99.7% homology with the Weissella koreensis S5623 16S rDNA (access no. AY035891). They were accordingly identified and named as Weissella koreensis OK1-4 and Weissella koreensis OK1-6, and could produce ornithine from MRS broth supplemented with 1% of arginine at a productivity of 27.01 and 31.41 mg/L/h, respectively. This is the first report on the production of ornithine by the genus Weissella isolated from kimchi.

Potential Probiotic Properties of Exopolysaccharide Producing Lactic Acid Bacteria Isolated from Fermented Soybean Product (장류유래 Exopolysaccharide 생성 유산균의 잠재적 Probiotic 특성)

  • Ahn, Yu-Jin;Choi, Hye-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.749-755
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    • 2014
  • Exopolysaccharides (EPSs) have been widely used in the food industry as viscofying, stabilizing, and emulsifying agents as well as in the pharmaceutical industry for their immunomodulatory, anti-tumor, and anti-inflammatory effects. A total of 458 lactic acid bacteria (LAB) strains isolated from several kinds of soybean pastes were screened for the production of homo-EPS (HoPS). LAB isolates were primarily screened using thin layer chromatography (TLC) and further screened polymerase chain reaction (PCR) targeting genes involved in HoPS production. Six LAB isolates producing high amounts of HoPS were identified by TLC. Among these isolates, glucansucrase gene was amplified in two strains (JSA57, JSB22), whereas the fructansucrase gene was detected in three strains (JSA57, JSB22, JSB66). After isolating the strains, their morphological characteristics and 16S rDNA sequences were determined. Six species were identified as L. alimentarius HSB15, L. plantarum JSA22, L. pentosus JSA57, L. brevis JSB22, L. alimentarius JSB66, and L. parabrevis JSB89. To evaluate the potential probiotic properties of these LAB, their survival rates against a simulated intestinal environment were determined. After 2 hr of incubation in artificial gastric juice, survival rates of JSA57, JSB90, JSB22, and JSB66 were all greater than 50%. After 2 hr of incubation in bile juice, viable cell count of JSB22 was similar with initial vial cell counts. Growth of the six LAB was screened in arabino-oligosaccharide (AOS)-containing MRS broth. Results showed that growth of the isolates selectively increased after culture in AOS-containing media. Strain JSB22 (6 hr), JSB66 (6 hr), HSB15 (20 hr), and JSA22 (29 hr) showed maximum growth rate. Especially, JSB22 showed the highest growth rate. These results suggest that EPS-producing LAB isolated from Deonjang could be applied as synbiotics.

Molecular Phylogeny of Chattonella (Raphidophyceae) Species from Deungnyang Bay, Korea Using Single-Cell PCR (Single-cell PCR을 이용하여 분석한 득량만 Chattonella 종 (Raphidophyceae)의 분자계통학적 특성)

  • Kim, Jin Joo;Song, Seon Yeung;Park, Tae Gyu
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.24 no.7
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    • pp.967-972
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    • 2018
  • The genus Chattonella belonging to the class raphidophyceae, is a harmful algal bloom species. Recently, its occurrence has been increasing and expanding along the Korean coast. Species identification of the genus Chattonella only by morphological observation is difficult due to the lack of rigid cell walls. In this study, the morphological characteristics and genetic affinity of Chattonella sp. isolated from Deungnyang Bay in 2017 were examined. We carried out single-cell isolation from field samples then sequenced three different areas using the single-cell PCR method: 1) parts of ribosomal operon, the large subunit (LSU) of the rDNA, 2) the chloroplast-encoded subunit psaA of Photosystem I, and 3) rbcL encoding the large subunit of the Rubisco gene. The cells were morphologically very similar to the general genus Chattonella ($74.0{\pm}10.1{\mu}m$ in length, $33.1{\pm}3.6{\mu}m$ in width). The three partial gene sequences were insufficient to justify distinction at the species rank. However, they clustered at 99-100 % sequence similarity with C. marina, C. marina var. antiqua and C. marina var. ovata.

Hypoglycemic Effect of Culture Broth of Bacillus subtilis S10 Producing 1-Deoxynojirimycin (1-Deoxynojirimycin을 생산하는 Bacillus subtilis S10 배양액의 혈당강하 효과)

  • Cho, Yong-Seok;Park, Young-Shik;Lee, Jae-Yeon;Kang, Kyung-Don;Hwang, Kyo-Yeol;Seong, Su-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1401-1407
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    • 2008
  • 1-Deoxynojirimycin (DNJ) is a strong $\alpha$-glucosidase inhibitor which inhibits hyperglycemia in animals. To select the Bacillus strains highly producing DNJ, 4,000 strains were isolated from soil and grain samples. By the inhibitory activity against $\alpha$-glucosidase, nine Bacillus strains were selected and then identified by 16S rDNA sequencing. B. subtilis S10 was finally selected as the best strain for the production of DNJ. Various carbon sources and nitrogen sources in culture medium were evaluated for the highest production of DNJ. As the results, the optimized concentration of carbon source and nitrogen source was 1.0% galactose and 1.6% polypeptone and the concentration of DNJ produced was 0.75 g/L. The effect of culture supernatant of B. subtilis S10 on lowering blood glucose level was investigated in streptozotocin (STZ)-induced diabetic mice model. Mice were randomly assigned to control group (saline) and three test groups such as acarbose group, silkworm powder group and B. subtilis S10 group. After eight-week oral feeding, blood glucose levels of the B. subtilis S10 and silkworm powder groups were respectively $209.1{\pm}19.6\;mg/dL$ (59.1%) and $208.6{\pm}39.8\;mg/dL$ (59.0%) lower than $510{\pm}10\;mg/dL$ of the control group. These results indicated that the culture supernatant of B. subtilis S10 was able to reduce the blood glucose level in STZ-induced diabetic mice.

Characteristics of Heterotrophic Bacterial Population in the Artificial Lake Geumgang Near Estuary Barrage (금강 하구둑 인근에서 미생물군집의 특성)

  • Bae, Myoung-Sook;Park, Suhk-Hwan;Choi, Gang-Guk;Lee, Keun-Kwang;Lee, Geon-Hyoung
    • The Korean Journal of Ecology
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    • v.28 no.3
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    • pp.129-134
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    • 2005
  • The monthly variations of physico-chemical and microbiological water quality were investigate in the artificial Lake Geumgang near estuary barrage. Sixty heterotrophic bacteria were isolated and identified by amplification and sequencing of 16S rDNA. Water temperature, pH, and inorganic nutrients($NH_4$-N, $NO_2$-N, $NO_3$-N, $PO_4$-P) were measured. Concentrations of DO, BOD, and inorganic nutrients were lower than in the middle-stream of Geum river The population densities of heterotrophic bacteria and total coliforms varied from $4.1{\pm}1.0\times10^2$ to $6.7{\pm}1.1{\times}10^3\;cfu\;ml^{-1}$, and 0 to $2.3{\pm}0.6{\times}10^2\;cfu\;ml^{-1}$, respectively. Among the measured numbers of physiological groups of bacteria, cellulolytic bacteria showed higher population densities than those of other physiological groups. Bacterial community structure was analysed based on 16S rDNA partial sequencing. Among 60 isolates, dominant genus was Pseudomones (20 strains).

Detection of Gram-negative Bacteria in Broad-range PCR Amplifying 16S rRNA Gene with Semi-nested Primers and Its Application in Market Milk (16S rRNA 유전자의 Semi-nested Primer를 이용한 Broad-range PCR에 의한 그람음성세균의 검출과 시유에서의 응용)

  • Choi, Suk-Ho;Choi, J.J.;Lee, S.B.
    • Journal of Animal Science and Technology
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    • v.47 no.3
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    • pp.465-474
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    • 2005
  • A two-step broad-range PCR method detecting gram-negative bacteria at the level as low as 2 CFU was developed by using primers of GNFI and GNRI and then semi-nested primer of GNF2 and GNRI. The nucleotide sequences of the primers were determined based on l6S rRNA gene. The DNA fragments of 1173 bp and 169 bp were amplified in one-step PCRs with primer sets of GNFI-GNRI and GNF2-GNRl, respectively, using template DNA from seven strains of gram-negative bacteria including Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas spp., and Acinetobacter baumaii but not from Achromobacter lyticus, Alca/igens faecalis, and five strains of gram-positive bacteria. DNA fragments of 180 bp were amplified from LTLT-pasteurized milk and UHf-pasteurized milk in the two-step PCR. The DNA fragments were amplified from LTLT-pasteurized milk which was added with Pseudomonas j/uorescens and subsequently heated at 65 $^{\circ}C$, 80 $^{\circ}C$, and 100 $^{\circ}C$ for 30 min but they were not amplified from the milk autoclaved at 121$^{\circ}C$ for 15 min. It was suggested in PCR that Pseudomonas fluorescens heated at 65 $^{\circ}C$ for 30 min in milk was more sensitive to DNase treatment than viable bacteria.

Antagonism against Helicobacter Pylori and Proteolysis of Lactobacillus Helveticus CU631 and Strain Identification

  • Yoon, Y.H.;Won, B.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1057-1065
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    • 2002
  • The antagonistic activities of 30 strains of lactobacilli against Helicobacter pylori were determined and Lactobacillus helveticus CU631 has been selected as the strain which possesses the strongest inhibitory effect in the disc diffusion assay showing inhibition zone diameter of $10{\pm}1.5mm$, whereas those of L. plantarum and L. fermentum have been shown to be $4.0{\pm}0.6mm$. H. pylori G88016 revealed the highest vacuolating toxin producing activity among the 8 strains, the inhibitory activity of L. helveticus CU631 in vacuolating toxin producing activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activities in urease and cytotoxin producing activities of H. pylori NCTC11637 and CJH12. An accelerated proteolytic activity of water soluble peptides by L. helveticus CU631 during the refrigeration storage has been manifested in the cream cheese. DNA seqences of 16S-23S ribosomal RNA spacer region showed typical pattern among the various strains of L. helveticus, which could be used in the identification of L. helveticus CU 631.

Variations of Hydrogen Production and Microbial Community with Different Nitrogen Concentration During Food Waste Fermentation (음식물쓰레기의 혐기성 소화 시 질소농도에 따른 수소생산 및 미생물 군집변화)

  • Lee, Pul-Eip;Lee, Tae-Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.10
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    • pp.672-678
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    • 2014
  • In this study, variations of fermentative hydrogen production and microbial community were investigated with different nitrogen concentration of food waste. Optimum hydrogen production rate was acquired at 200 mg/L nitrogen concentration of the food waste. Which was eqivalent to 83.43 mL/g dry biomass/hr. However, bio-hydrogen production was inhibitedly reduced at over 600 mg/L of nitrogen concentration whereas proportional relation between hydrogen production and B/A ratio were not observed. Most dominant specie of the microbial community analyzed was Clostridium sp. throughout PCR-DGGE analysis of 16S rDNA. It revealed that most contributing microorganism producing hydrogen were Enterococcus faecium partial, Klebsiella pneumoniae strain ND6, Enterobacter sp. NCCP-231, and Clostridium algidicarnis strain E107 in this experiment.

Molecular Characterization of Burkholderia Strains Isolated from Rice Cultivars (Oryza sativa L.) for Species Identification and Phylogenetic Grouping

  • Madhaiyan, Munusamy;Poonguzhali, Selvaraj;Kwon, Soon-Wo;Song, Myung-Hee;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1005-1010
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    • 2008
  • The genus Burkholderia consists of extremely versatile bacteria that occupy diverse niches and are commonly encountered in the rhizosphere of crop plants. In this study, we characterized three plant growth promoting strains assigned as Burkholderia sp. using biochemical and molecular characterization. The Burkholderia spp. strains CBMB40, CBPB-HIM, and CBPB-HOD were characterized using biochemical tests, BIOLOG carbon substrate utilization, fatty acid methyl ester analysis, analysis of recA gene sequences, and DNA-DNA hybridization. The results from these studies indicated that the strains CBMB40, CBPB-HIM, and CBPB-HOD can be assigned under Burkholderia vietnamiensis, Burkholderia ubonensis, and Burkholderia pyrrocinia, respectively.

Biodegradation of Kerosene by Pseudomonas aeruginosa K14 (Pseudomonas aeruginosa Kl4를 이용한 등유(Kerosene)의 생물학적 분해)

  • Kim, Jee-Young;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.44 no.2
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    • pp.156-163
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    • 2008
  • In this study, we isolated 32 strains of kerosene degrading bacteria from oil contaminated soil by enrichment culture. Isolates were screened for kerosene degradation efficiencies and K14 were selected which had the highest removal efficiency for 1,000 mg/L of kerosene. K14 were identified as Pseudomonas aeruginosa by morphological, biochemical test and 16S rDNA analysis. The optimal culture condition were determined as initial inoculated cell concentration, 1.0 g/L; substrate concentration, 1,000 mg/L; temperature $30^{\circ}C$; pH 7. When we enforced batch test in this condition, K14 degraded 72% of kerosene with 1,000 mg/L during 72 hr. And, at low concentration (200 mg/L), K14 degraded 95.8% of kerosene during 48 hr. As a result, kerosene biodegradation by Pseudomonas aeruginosa K14 could be useful for clean up of groundwater and soil contaminated with crude oil.