• Title/Summary/Keyword: 16S-rRNA

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Microbial Diversity of the Trichloroethylene Contaminated Groundwater Treatment System and Characterization of Pseudomonas sp. DHC8 (Trichloroethylene으로 오염된 지하수 제거공정의 미생물 다양성 및 분리균주 Pseudomonas sp. DHC8의 특성)

  • Nam, Ji-Hyun;Shin, Ji-Hye;Kwon, Kiwook;Bae, Wookeun;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.336-342
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    • 2013
  • Trichloroethylene (TCE) is a widely used substance in commercial and industrial applications, yet it must be removed from the contaminated soil and groundwater environment due to its toxic and carcinogenic nature. We investigated bacterial community structure, dominant bacterial strain, and removal efficiency in a TCE contaminated groundwater treatment system using immobilized carrier. The microbial diversity was determined by the nucleotide sequences of 16S rRNA gene library. The major bacterial population of the contaminated groundwater treatment system was belonging to BTEX degradation bacteria. The bacterial community consisted mainly of one genus of Pseudomonas (Pseudomonas putida group). The domination of Pseudomonas putida group may be caused by high concentration of toluene and TCE. Furthermore, we isolated a toluene and TCE degrading bacterium, named Pseudomonas sp. DHC8, from the immobilized carrier in bioreactor which was designed to remove TCE from the contaminated ground water. Based on the results of morphological and physiological characteristics, and 16S rRNA gene sequence analysis, strain DHC8 was identified as a member of Pseudomonas putida group. When TCE (0.83 mg/L) and toluene (60.61 mg/L) were degraded by this strain, removal efficiencies were 72.3% and 100% for 12.5 h, respectively. Toluene removal rate was 2.89 ${\mu}mol/g$-DCW/h and TCE removal rate was 0.02 ${\mu}mol/g$-DCW/h. These findings will be helpful for maintaining maximum TCE removal efficiency of a reactor for bioremediation of TCE.

Studies on the Functional Properties of Lactic Acid Bacteria Isolated from Home-made Yogurt and Commercial Yogurt (Home-made 요구르트와 시판 중인 요구르트에서 분리한 젖산균의 기능적 특성 조사)

  • Choi, Moon-Sup;Yun, Hyun-Myoung;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.8-14
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    • 2014
  • The objective of this work is to investigate and compare several functional properties of lactic acid bacteria (LAB), Lactobacillus casei SK-7 isolated from home-made yogurt and Lactobacillus bulgaricus YK-11 from commercial yogurt. Initially, physiological and biochemical properties of SK-7 and YK-11 were characterized. Phylogenetic analysis using 16S rRNA sequencing were performed to identify the strains, and the strain could be assigned to Lactobacillus casei and Lactobacillus bulgaricus, designated as L. casei SK-7 and L. bulgaricus YK-11. Phylogenetic tree of SK-7 and YK-11 was plotted based on 16S rRNA sequence comparisons. Production of lactic acid and organic acid, and pH changes in the cultures of SK-7 and YK-11 were monitored during 72 h. During the incubation period, several functional properties of L. casei SK-7 and L. bulgaricus YK-11 were examined. L. casei SK-7 and L. bulgaricus YK-11 cultures eliminated 93.9% and 88.2% of nitrite, respectively. Antioxidant activity of cultural supernatants of SK-7 and YK-11 were 62.6%, 54.9%, and activity of ${\beta}$-galactosidase were 14.9 units/mg and 13.1 units/mg, respectively. The antimicrobial activities were examined with 20-fold concentrated culture supernatants from the cultures of SK-7 and YK-11. The activities of SK-7 supernatants were clearly observed against all microorganisms in this work, whereas no activities were observed in YK-11 supernatants. Although it might be conducted additional functional research, functional properties of LAB isolated from home-made yogurt have been shown to be better than those of commercial yogurt in this work.

Population Genetic Structure of Octopus minor Sasaki from Korea and China Based on a Partial Sequencing of Mitochondrial 16S rRNA (미토콘드리아 16S rRNA 염기서열에 의한 한국, 중국 낙지의 유전자 집단 분석)

  • Kim, Joo-Il;Oh, Taeg-Yun;Seo, Young-Il;Cho, Eun-Seob
    • Journal of Life Science
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    • v.19 no.6
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    • pp.711-719
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    • 2009
  • We determined a portion of mitochondrial 16S rRNA gene sequences (416 bp) to investigate the genetic structure of the octopus (Octopus minor Ssaki) population in Korea and China. Samples were obtained from Korea (Yeosu, Namhae, Jindo, Muan, Geomundo and Seosan) and China (Sandong) during the period of August 2006 to September 2007. Sequence analyses of 28 individual specimens collected from 7 localities revealed 11 haplotypes, ranging in a sequence divergence of 0.2% - 1.2%. Phylogenetic analyses using PHYLIP and networks subdivided the octopus into two clades (termed clade A and B) and the nucleotide divergence between them was 0.4%. This haplotype subdivision was in accordance with geographic separation: one at Yeosu, Namhae, Muan and Jindo, and the other at Seosan, Geomundo and Sandong. On the basis of hierarchial genetic analysis, genetic distance between localities in Korea and China were also found, but a significant population differentiation was not shown in this study (p>0.05). Consequently, most of the octopus populations in Korea had considerable distribution due to the mitochondrial gene flow that resulted in a formation of a genetically homogenous structure, whereas some of the Korean and Chinese populations had different genetic structures. Gene flow among populations may be restricted due to impassable geographic barriers that promote genetic differentiation.

Change of Gene Expression Pattern of Mycobacterium tuberculosis H37Rv Against Host Immune Response in Infected Mouse Lung (결핵균 H37Rv에 감염된 마우스의 폐에서 면역 반응에 대항하는 Mtb 유전자의 발현 변화)

  • Lee, Hyo-Ji;Cho, Jung-Hyun;Kang, Su-Jin;Jung, Yu-Jin
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.134-139
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    • 2010
  • Mycobacterium tuberculosis (Mtb) is one of the most successful pathogens to infect one third of world population. Th1-mediated immunity against Mtb infection is known as critical to express mycobacteriostatic function but it is not sufficient to resolve the infection. In this study, to verify the possibility Mtb itself change the gene expression to survive against host immune response, expression pattern of selected H37Rv genes, 16S rRNA, acr, fbpA, aceA, and ahpC, during the course of infection was measured with absolute quantitation method using real-time RT-PCR. The total number of transcripts of 16S rRNA increased during the course of infection, which was coincide with the increasing CFU. The total number of fbpA transcripts per CFU, which encode typical secreted Mtb antigen, Ag85A, increased for 10 days of infection before decreasing. The number of transcripts of acr per CFU, which encode heat shock protein, ${\alpha}$-crystallin, increased during the infection, and ahpC and aceA, they both are enzymes produced in oxidative stressful condition, increased for 20 days and then slightly decreased on day 30. These findings are one of survival strategy of pathogen evading host immune response lead to persistent infection inside host cells.

Isolation and Physiological Characteristics of Microorganisms Producing Extracellular Enzymes from Korean Traditional Soybean Sauce and Soybean Paste (전통 장류에서 세포외효소 분비능이 우수한 미생물의 분리 및 생리활성 특성)

  • Baek, Seong-Yeol;Yun, Hye-Ju;Choi, Hye-Sun;Koo, Bon-Sung;Yeo, Soo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.379-384
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    • 2010
  • We isolated microorganisms presenting high enzymatic activities for amylase, cellulase, protease, lipase or fibrinolysis from Korean traditional soybean sauce and paste. Then, the physiological properties and 16S rRNA sequences of isolated microorganisms were analyzed. All of the isolated 13 strains possessing high extra cellular enzyme activities have higher amylase and cellulase activities than Bacillus subtilis KACC 10114. All the selected strains have protease activities except for D2-14. Except D8-8 and K4-1, other strains have lipase activity. D2-7, D8-8 and K4-1 strains have higher fibrinolytic activities than others, while D8-2 strain has no activity. Most of the selected strains showed antibacterial activity even in gram positive and gram negative bacteria and yeast. Gene sequence analysis of 16S rRNA from isolated strains revealed that all the selected strains were member of Bacillus species.

Biphasic Study to Characterize Agricultural Biogas Plants by High-Throughput 16S rRNA Gene Amplicon Sequencing and Microscopic Analysis

  • Maus, Irena;Kim, Yong Sung;Wibberg, Daniel;Stolze, Yvonne;Off, Sandra;Antonczyk, Sebastian;Puhler, Alfred;Scherer, Paul;Schluter, Andreas
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.321-334
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    • 2017
  • Process surveillance within agricultural biogas plants (BGPs) was concurrently studied by high-throughput 16S rRNA gene amplicon sequencing and an optimized quantitative microscopic fingerprinting (QMF) technique. In contrast to 16S rRNA gene amplicons, digitalized microscopy is a rapid and cost-effective method that facilitates enumeration and morphological differentiation of the most significant groups of methanogens regarding their shape and characteristic autofluorescent factor 420. Moreover, the fluorescence signal mirrors cell vitality. In this study, four different BGPs were investigated. The results indicated stable process performance in the mesophilic BGPs and in the thermophilic reactor. Bacterial subcommunity characterization revealed significant differences between the four BGPs. Most remarkably, the genera Defluviitoga and Halocella dominated the thermophilic bacterial subcommunity, whereas members of another taxon, Syntrophaceticus, were found to be abundant in the mesophilic BGP. The domain Archaea was dominated by the genus Methanoculleus in all four BGPs, followed by Methanosaeta in BGP1 and BGP3. In contrast, Methanothermobacter members were highly abundant in the thermophilic BGP4. Furthermore, a high consistency between the sequencing approach and the QMF method was shown, especially for the thermophilic BGP. The differences elucidated that using this biphasic approach for mesophilic BGPs provided novel insights regarding disaggregated single cells of Methanosarcina and Methanosaeta species. Both dominated the archaeal subcommunity and replaced coccoid Methanoculleus members belonging to the same group of Methanomicrobiales that have been frequently observed in similar BGPs. This work demonstrates that combining QMF and 16S rRNA gene amplicon sequencing is a complementary strategy to describe archaeal community structures within biogas processes.

Analysis of 16S rRNA gene sequencing data for the taxonomic characterization of the vaginal and the fecal microbial communities in Hanwoo

  • Choi, Soyoung;Cha, Jihye;Song, Minji;Son, JuHwan;Park, Mi-Rim;Lim, Yeong-jo;Kim, Tae-Hun;Lee, Kyung-Tai;Park, Woncheoul
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1808-1816
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    • 2022
  • Objective: The study of Hanwoo (Korean native cattle) has mainly been focused on meat quality and productivity. Recently the field of microbiome research has increased dramatically. However, the information on the microbiome in Hanwoo is still insufficient, especially relationship between vagina and feces. Therefore, the purpose of this study is to examine the microbial community characteristics by analyzing the 16S rRNA sequencing data of Hanwoo vagina and feces, as well as to confirm the difference and correlation between vaginal and fecal microorganisms. As a result, the goal is to investigate if fecal microbiome can be used to predict vaginal microbiome. Methods: A total of 31 clinically healthy Hanwoo that delivered healthy calves more than once in Cheongju, South Korea were enrolled in this study. During the breeding season, we collected vaginal and fecal samples and sequenced the microbial 16S rRNA genes V3-V4 hypervariable regions from microbial DNA of samples. Results: The results revealed that the phylum-level microorganisms with the largest relative distribution were Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria in the vagina, and Firmicutes, Bacteroidetes, and Spirochaetes in the feces, respectively. In the analysis of alpha, beta diversity, and effect size measurements (LefSe), the results showed significant differences between the vaginal and fecal samples. We also identified the function of these differentially abundant microorganisms by functional annotation analyses. But there is no significant correlation between vaginal and fecal microbiome. Conclusion: There is a significant difference between vaginal and fecal microbiome, but no significant correlation. Therefore, it is difficult to interrelate vaginal microbiome as fecal microbiome in Hanwoo. In a further study, it will be necessary to identify the genetic relationship of the entire microorganism between vagina and feces through the whole metagenome sequencing analysis and meta-transcriptome analysis to figure out their relationship.

Postsurgical Wound Infection Caused by Mycobacterium conceptionense Identified by Sequencing of 16S rRNA, hsp65, and rpoB Genes in an Immunocompetent Patient (16S rRNA, hsp65, 및 rpoB 염기순서분석으로 동정한 Mycobacterium conceptionense에 의한 면역능이 정상인 환자에서 발생한 수술후 창상감염)

  • Lee, Ja Young;Kim, Si Hyun;Shin, Jeong Hwan;Lee, Hyun-Kyung;Lee, Young Min;Song, Sae Am;Bae, Il Kwon;Kim, Chang-Ki;Jun, Kyung Ran;Kim, Hye Ran;Lee, Jeong Nyeo;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.17 no.1
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    • pp.23-27
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    • 2014
  • Rapidly growing mycobacteria are ubiquitous in the environment and are increasingly being recognized as opportunistic pathogens. Recently, a new species, Mycobacteium conceptionense, has been validated from the Mycobacterium fortuitum third biovariant complex by molecular analysis. However, there are few reports, and postsurgical wound infection by this species is rare. We report a case of postsurgical wound infection caused by M. conceptionense in an immunocompetent patient that was identified by a sequencing analysis of 16S rRNA, hps65, and rpoB genes.

Development of Recombinant Escherichia coli Expressing Rhodotorula glutinis Epoxide Hydrolase (Rhodotorula glutinis의 epoxide hydrolase 고효율 발현 유전자 재조합 Escherichia coli 생촉매 개발)

  • Lee Soo-Jung;Kim Hee-Sook
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.415-419
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    • 2006
  • The epoxide hydrolase (EH) of Rhodotorula glutinis which has a high enantioselectivity against aromatic epoxide substrates was expressed to high levels in Escherichia coli based on codon usage. We analysed the Preference of codon usage between the yeast, R. glutinis, and bacteria, E. coli. E. coli, Rosetta(DE3)pLysS, harbors pRARE plasmid with tRNA genes for rare-codons was employed as a host strain. The recombinant E. coli expressing R. glutinis EH showed an enhanced enantioselective hydrolysis activity toward racemic styrene oxide. Enantiopure (S)-styrene oxide with a high enantiopurity of 99% ee (enantiomeric excess) was obtained from racemic substrates.

Inhibitory Effects on Melanin Production in B16 Melanoma Cells of Sedum sarmentosum (B16 Melanoma 세포에서 돌나물 추출물의 멜라닌 생성 저해 효과)

  • Sim, Gwan-Sub;Kim, Jin-Hwa;Lee, Bum-Chun;Lee, Dong-Hwan;Lee, Geun-Soo;Pyo, Hyeong-Bae
    • YAKHAK HOEJI
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    • v.52 no.3
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    • pp.165-171
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    • 2008
  • To develop effective skin whitening agents, we tested natural herbal extracts for their melanogenic inhibitory activities. Sedum samentosum was selected for its inhibitory effect on melanogenesis in B16 melanoma cells. Ethanolic extract of S. samentosum (SSE) was evaluated for antioxidative effect and tyrosinase inhibitory activity of melanogenesis. We investigated the changes in protein level and mRNA level of tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 by using western blotting and RT-PCR, respectively. SSE showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}$ of 342.7 $\mug/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 64.69 $\mug/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. SSE treatment suppressed the biosynthesis of melanin up to 46% and reduced tyrosinase activity up to 51% at 100 $\mug/ml$ in B16 melanoma cells. The tyrosinase activity and tyrosinase expression in B16 melanoma cells were reduced in a dose-dependent manner by SSE. Also, SSE was able to significantly inhibit tyrosinase and TRP-1 expression in mRNA level. These results suggest that SSE inhibited melanin production which may be dependent on tyrosinase activity and expression in B16 melanoma cells, and an effective whitening agent for the skin.