• Title/Summary/Keyword: rDNA sequencing

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Identification of Stenotrophomonas maltophilia LK-24 and its Degradability of Crystal Violet

  • Kim, Jeong-Dong;Yoon, Jung-Hoon;Park, Yong-Ha;Fusako Kawai;Kim, Hyun-Tae;Lee, Dae-Weon;Kang, Kook-Hee
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.437-443
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    • 2002
  • A number of soil and wastewater samples were collected from the vicinity of an effluent treatment plant for the chemical industry. Several microorganisms were screened fur their ability to decolorize the triphenylmethane group of dyes. As a result, a novel crystal violet dye-degrading strain LK-24 was isolated. Taxonomic identification including 16S rDNA sequencing and phylogenetic analysis indicated that the isolate had a $99.5\%$ homology in its 16S rDNA base sequence with Stenotrophomonas maltophilia. The triphenylmethane dye, crystal violet, was degraded extensively by growing cells of Stenotrophomonas maltophilia LK-24 in agitated liquid cultures, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly degraded at a relatively lower concentration, below $100{\mu}g\;ml^-1$, yet the growth of the cells was totally suppressed at a dye concentration of $250{\mu}g\;ml^-1$. The degradation products of crystal violet were identified as 4,4'-bis(dimethylamino)-benzophenone and ${\rho}$-dimethylaminophenol by Gas chromatography-Mass spectrometry. The 4,4'-bis(dimethylamino)-benzophenone was easily obtained in a reasonable yield, as it was not metabolized further by S. maltophilia LK-24; however, the ${\rho}$-dimethylaminophenol was not easily identifiable, as it was further metabolized.

Monitoring the Ecology of Bacillus During Daqu Incubation, a Fermentation Starter, Using Culture-Dependent and Culture-Independent Methods

  • Yan, Zheng;Zheng, Xiao-Wei;Han, Bei-Zhong;Han, Jian-Shu;Nout, M.J. Robert;Chen, Jing-Yu
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.614-622
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    • 2013
  • Daqu, a traditional fermentation starter, has been used to produce attractively flavored foods such as vinegar and Chinese liquor for thousands of years. Although Bacillus spp. are one of the dominant microorganisms in Daqu, more precise information is needed to reveal why and how Bacillus became dominant in Daqu, and next, to assess the impact of Bacillus sp. on Daqu and its derived products. We combined culture-dependent and culture-independent methods to study the ecology of Bacillus during Daqu incubation. Throughout the incubation, 67 presumptive Bacillus spp. isolates were obtained, 52 of which were confirmed by 16S rDNA sequencing. The identified organisms belonged to 8 Bacillus species: B. licheniformis, B. subtilis, B. amyloliquefaciens, B. cereus, B. circulans, B. megaterium, B. pumilus, and B. anthracis. A primer set specific for Bacillus and related genera was used in a selective PCR study, followed by a nested DGGE PCR targeting the V9 region of the 16S rDNA. Species identified from the PCR-DGGE fingerprints were related to B. licheniformis, B. subtilis, B. amyloliquefaciens, B. pumilus, B. benzoevorans, and B. foraminis. The predominant species was found to be B. licheniformis. Certain B. licheniformis strains exhibited potent antimicrobial activities. The greatest species diversity occurred at the Liangmei stage of Daqu incubation. To date, we lack sufficient knowledge of Bacillus distribution in Daqu. Elucidating the ecology of Bacillus during Daqu incubation would enable the impact of Bacillus on Daqu to be accessed, and the quality and stabilization of Daqu-derived products to be optimized.

Association of a miR-502-Binding Site Single Nucleotide Polymorphism in the 3'-Untranslated Region of SET8 and the TP53 Codon 72 Polymorphism with Cervical Cancer in the Chinese Population

  • Yang, Shao-Di;Cai, Yan-Lin;Jiang, Pei;Li, Wen;Tang, Jian-Xin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6505-6510
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    • 2014
  • Objective: This study was conducted to identify whether polymorphic variants of set domain-containing protein 8 (SET8) and tumor protein p53 (TP53) codon 72, either independently or jointly, might be associated with increased risk for cervical cancer. Methods: We genotyped SET8 and TP53 codon 72 polymorphisms of peripheral blood DNA from 114 cervical cancer patients and 200 controls using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct DNA sequencing. Results: The frequency of SET8 CC (odds ratios (OR) = 2.717, 95% CI=1.436-5.141) or TP53 GG (OR=2.168, 95% CI=1.149-4.089) genotype was associated with an increased risk of cervical cancer on comparison with the SET8 TT or TP53 CC genotypes, respectively. In additional, interaction between the SET8 and TP53 polymorphisms increased the risk of cervical cancer in a synergistic manner, with the OR being 9.913 (95% CI=2.028-48.459) for subjects carrying both SET8 CC and TP53 GG genotypes. Conclusion: These data suggest that there are significant associations between the miR-502-binding site SNP in the 3'-UTR of SET8 and the TP53 codon 72 polymorphism with cervical cancer in Chinese, and there is a gene-gene interaction.

Characterization of Growth Inhibition and Isolation of a Copper-Resistant Rhizobacterium, Alcaligenes sp. KC-1 (Cu 내성 근권 세균 Alcaligenes sp. KC-1의 분리 및 생장특성)

  • Hong, Sun-Hwa;Shin, Ki-Chul;Lee, Eun-Young
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.182-187
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    • 2011
  • In this study, A bacterium with an ability to resist toxic heavy metals was isolated from reeds in wetland. The isolated strain was identified to Alcaligenes sp. KC-1 by 16S rDNA sequencing. Heavy metals such as Pb, $Cr^{6+}$, Cd, Zn and Cu were supplied to media. The ecotoxic treat of the heavy metals on the growth of strain KC-1 was performed when the isolated strain Alcaligenes sp. KC-1 cultured with Cu ranging from 0 mM to 20 mM. It showed the resistance of $EC_{50}$(7.34 mM) and cell growth ($OD_{600\;nm}$ : 0.83 after 42 hours) when it was cultured in Cu.

Study on the promotion of inflammation and whitening of natural materials using bioconversion technology

  • Lee, Se-Won;Lim, Jeong-Muk;Lee, Seong-Hyeon;Lee, Jeong-Ho;Oh, Byung-Teak
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.116-116
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    • 2018
  • Bioconversion, the enzymatic process by microorganism on organic precursor to desired products. The natural extract is converted into a form that can be easily absorbed into the skin, while scaling up of to higher quantity is possible. Selection of naturally processed raw material rather than chemically processed is preferred. Therefore, fermentation was carried out by mixing Rubus coreanus Miquel, soybean, Zanthoxylum schinifolium as bioconverting materials, the possibility of inflammation, whitening material were checked. In this study, useful microorganisms were isolated from various salted fish, and 16S rDNA sequence was analyzed to confirm their genetic characteristics. Combining the three natural materials using bioconversion technology to study their activity before and after fermentation. To evaluate the antioxidant activity and the active ingredient content the DPPH radical scavenging activity and the total polyphenol content were examined. Raw 264.7 cells were used to evaluate MTT assay, NO and $TNF-{\alpha}$ production inhibitory activity. Also, to evaluate the whitening activity, tyrosinase inhibitory activity and melanin formation inhibitory activity were measured using B16F10 cells. In total 34 strains were obtained from various salted fish. The effective strains useful for the bioconversion process, showed that DPPH radical scavenging ability and polyphenol content were increased in the two kinds of microbial treatment groups compared to the untreated group. 16S rDNA sequencing analysis of the strains showed excellent in Pediococcus pentosaceus B1 in comparison. An increase of up-to 156% in anti-oxidative activity and 141% in polyphenol content was observed after bioconversion. In addition, after mixed fermentation the toxidty of Raw 264.7 and B16F10 cells tended to decrease and a significant increase was observed in anti-inflammatory activity as well. Also, tyrosinase activity and melanin significantly. synthesis decreased significantly.

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Chemical Characteristics and Biological Activities of Herbimycin A and Dihydroherbimycin A Produced by a Soil Isolate Streptomyces sp. AO-0511 (Streptomyces sp. AO-0511이 생산하는 Herbimycin A 및 Dihydroherbimycin A의 이화학적 특성 및 생물 활성)

  • Chang, Hung-Bae;Kim, Se-Chan;Kim, Jae-Heon
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.47-53
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    • 2006
  • A streptomycete strain was isolated from the soil samples from Korea. The chemotaxonomy and 16S rDNA sequencing confirmed that the strain belonged to the genus Streptomyces and we named it Streptomyces sp. AO-0511. Two antibiotics, herbimycin A and dihydroherbimycin A produced by this strain were tested for their physico-chemical and biological characteristics. Both compounds were stable under acidic pH. Dihydroherbimycin A was more heat-stable and polar compared with herbimycin A. Only weak antibacterial activities were detected against Bacillus subtilus ATCC 6633 and Micrococcus luteus ATCC 9341. However, herbimycin A and dihydroherbimycin A showed strong inhibitory activities on lung cancer cells (A549 cells) and leukemia cells (HL-60). The cytotoxicity was determined using L5178Y and P388 cell lines. The results showed that herbimycin A and dihydroherbimycin A had lower toxic effects on the cells compared with the standard compounds, comptothecin and cyclosporin A. Therefore, both compounds could be good candidates for the development of new anticancer drugs.

Structure and Characteristics of Bacterial Community on Biofilm of Food Wastewater Treatment System in Winter (식품폐수 처리 공정용 생물막의 겨울철 세균군집 구조와 특성)

  • Lee, Dong-Geun;Yoo, Ki-Hwan;Park, Seong-Joo
    • Journal of Environmental Health Sciences
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    • v.37 no.2
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    • pp.124-132
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    • 2011
  • Biofilm and aeration tank of pilot and full RABC (rotating activated Bacillus contactor) plant were analyzed to characterize and determine bacterial community structure in food wastewater treatment system at winter. Concentration of heterotrophic bacteria and Bacillus group was $10^7$ and $10^5$ CFU/ml, respectively, at biofilm of pilot-plant while others represented $10^6$ and $10^4$ CFU/ml, respectively. Five and eight phyla were detected at biofilm of pilot- and full-plant, respectively, by 16S rDNA sequencing. Biofilm of pilot-plant was dominated by ${\beta}$-Proteobacteria (38.8%), ${\gamma}$-Proteobacteria (22.4%), and Bacteroidetes (12.2%), and the most dominant genus was Zoogloeae genus (22.4%). Candidate division TM7 (12.5%) was only detected at biofilm of full-plant and it was dominated by Bacteroidetes (33.3%), ${\gamma}$-Proteobacteria (29.2%), and ${\beta}$-Proteobacteria (20.8%). Clostridium genus specific primer set enabled to detect the sequences of Clostridium genus. These suggested that anaerobic and aerobic bacteria were coexisted even from the initial period of biofilm formation and ${\beta}$-Proteobacteria, ${\gamma}$-Proteobacteria and Bacteroidetes were major phyla in biofilm of food wastewater treatment system at winter.

Expression of Recombinant Korean Mistletoe(KM) Lectin and B genes in Saccharomyces cerevisiae (Saccharomyces cerevisae에서 한국산 겨우살이 유래 lectin A 및 B 유전자의 발현)

  • 최윤혁;김종배;양웅석;황철원
    • Journal of Life Science
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    • v.14 no.5
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    • pp.840-846
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    • 2004
  • A study for expression of Korean Mistletoe (KM) lectin gene (A,B) in Saccharomyces cerevisiae was done using transforming system of yeast. In order to overexpress the genes efficiently in yeast, two lectin genes (A,B) were re-cloned and modified including Kozak translation initiation sequence using PCR amplification. The constructed plasmids containing modified lectin A and B genes were transformed to S. cerevisea INVSc (MAT G, his3 $\Delta$1, leu2, trpl-289, ura3-52). The transformed cells were identified by DNA sequencing with ABI3700 system and induced with 2% of galactose for recombinant KM lectin (rKM lectin) protein. The rKM lectin A and B proteins were determinated about 29kDa size of protein by SOS-P AGE and western blotting analysis. The expressed recombinant lectin was determinated 1.24∼1.75 $\mu\textrm{g}$ per 1 mg of cytosolic soluble protein by sandwich ELISA method. Moreover the lectin genes were expressed as maximum level at 36 h after galactose induction and lectin A gene was were repressed after 48 h.

Isolation and Identification of a Lactic Acid Bacterial Strain KJ-108 and Its Capability for Deodorizing Malodorous Gases Under Anaerobic Culture Conditions

  • KIM, JEONG-DONG;JUNG-HOON YOON;YONG-HA PARK;DAE-WEON LEE;KYOU-SEUNG LEE;CHANG-HYUN CHOI;WON-YEOP PARK;KOOK-HEE KANG
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.207-216
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    • 2003
  • A number of different sources, such as composts, leachates, and pig feces samples, were collected from different pig farms in Korea, and several microorganisms were screened for their ability to deodorize the malodorous gases. Consequently, a novel malodorous gases-deodorizing bacterial strain, KJ-108. was isolated, because it was highly abundant in nitrate-supplemented minimal medium ($MM-NO_3^-$) under anaerobic culture conditions. Airtight crimp-sealed serum bottles containing $MM-NO_3^-$ , medium were inoculated with KJ-108. Nitrate concentration was decreased rapidly after 20 h of incubation, and incubation was carried out until nitrite production reached almost zero. Taxonomic identification, including 16S rDNA base sequencing and phylogenetic analysis, indicated that the isolate had $100\%$ homology in its 165 rDNA base sequence with Lactobacillus pentosus. Among the volatile fatty acids, acetic acid contained in large amounts in fresh piggery slurry was decreased by about $40\%$ after 50 h incubation with strain KJ-108. n-Butyric acid, n-valeric acid, and isovaleric acid were gradually decreased, and isobutyric acid and capronic acid were dramatically eliminated at theinitial period with the treatment. Moreover, NH, removal efficiency reached a maximum of $98.5\%$ after 50 h of incubation, but the concentration of $H_2S$ was not changed.

Molecular Phylogenetic Diversity and Spatial Distribution of Bacterial Communities in Cooling Stage during Swine Manure Composting

  • Guo, Yan;Zhang, Jinliang;Yan, Yongfeng;Wu, Jian;Zhu, Nengwu;Deng, Changyan
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.6
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    • pp.888-895
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    • 2015
  • Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and subsequent sub-cloning and sequencing were used in this study to analyze the molecular phylogenetic diversity and spatial distribution of bacterial communities in different spatial locations during the cooling stage of composted swine manure. Total microbial DNA was extracted, and bacterial near full-length 16S rRNA genes were subsequently amplified, cloned, RFLP-screened, and sequenced. A total of 420 positive clones were classified by RFLP and near-full-length 16S rDNA sequences. Approximately 48 operational taxonomic units (OTUs) were found among 139 positive clones from the superstratum sample; 26 among 149 were from the middle-level sample and 35 among 132 were from the substrate sample. Thermobifida fusca was common in the superstratum layer of the pile. Some Bacillus spp. were remarkable in the middle-level layer, and Clostridium sp. was dominant in the substrate layer. Among 109 OTUs, 99 displayed homology with those in the GenBank database. Ten OTUs were not closely related to any known species. The superstratum sample had the highest microbial diversity, and different and distinct bacterial communities were detected in the three different layers. This study demonstrated the spatial characteristics of the microbial community distribution in the cooling stage of swine manure compost.