• Title/Summary/Keyword: Activity Sequencing

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혐기성 SBR을 이용한 anammox 미생물 배양 및 fluorescence in situ hybridization (FISH)을 통 미생물 군집 분석

  • Han, Dong-U;Yun, Ho-Jun;Kim, Dong-Jin
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.286-289
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    • 2001
  • Anaerobic ammonium oxidation with nitrite to $N_2$(anammox) is a recently discovered microbial reaction with interesting potential for nitrogen removal from wastewater. Here we investigated the microbial community structure in the sequencing batch reactor(SBR) with an anammox activity. The SBR was optimized for the enrichment of a very slowly growing microbial community and showed that possibility of anaerobic ammonium oxidation. Fluorescence in situ hybridization(FISH) analysis revealed that anaerobic ammonium oxidizers were Candidatus Brocadia anammoxidans and Candidatus Kuenenia stuttgartiensis. Furthermore, Nitrosomol1as spp. of the ${\beta}$ -subclass of Proteobacteria was also present within the anaerobic SBR microorganisms.

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Purification, Characterization, and Cloning of Trimethylamine Dehydrogenase from Methylophaga sp. Strain SK1

  • Kim, Hee-Gon;Kim, Yan;Lim, Heon-Man;Shin, Hyun-Jae;Kim, Si-Wouk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.337-343
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    • 2006
  • Trimethylamine dehydrogenase (TMADH, EC 1.5.99.7), an iron-sulfur flavoprotein that catalyzes the oxidative demethylation of trimethylamine to form dimethylamine and formaldehyde, was purified from Methylophaga sp. strain SK1. The active TMADH was purified 12.3-fold through three purification steps. The optimal pH and temperature for enzyme activity was determined to be 8.5 and $55^{\circ}C$, respectively. The $V_{max}\;and\;K_m$ values were 7.9 nmol/min/mg protein and 1.5 mM. A genomic DNA of 2,983 bp from Methylophaga sp. strain SK1 was cloned, and DNA sequencing revealed the open reading frame (ORF) of the gene coding for TMADH. The ORF contained 728 amino acids with extensive identity (82%) to that of Methylophilus methylotrophus $W_3A_1$.

Rapid Screening of Mutant Strains of Trigonopsis variabilis (ATCC10679) for Cephalosporin C Bioconversion and Sequences of D-amino acid oxidase Genes (Cephalosporin C 생물전환을 위한 Trigonopsis variabilis (ATCC10679) 변이균주의 간편한 선별 및 D-amino acid oxidase 유전자 배열)

  • 강용호;박선영
    • KSBB Journal
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    • v.14 no.2
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    • pp.235-240
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    • 1999
  • Simple and rapid screening methods were developed to screen mutant strains of Trigonopsis variabilis ATCC10679 (TW). D-amino acid oxidase (D-AAO) from a mutant strain, T26, showed about 30% higher specific activity against cephalosporin C than from its wild type, TW. D-AAO genes from both TW and T26 strains were cloned and sequenced. There was one nucleotide changed from T to C at 811 position, resulting in an amino acid codon changed from Phe-258 to Ser-258.

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DNA Light-strand Preferential Recognition of Human Mitochondria Transcription Termination Factor mTERF

  • Nam, Sang-Chul;Kang, Chang-Won
    • BMB Reports
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    • v.38 no.6
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    • pp.690-694
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    • 2005
  • Transcription termination of the human mitochondrial genome requires specific binding to termination factor mTERF. In this study, mTERF was produced in E. coli and purified by two-step chromatography. mTERF-binding DNA sequences were isolated from a pool of randomized sequences by the repeated selection of bound sequences by gel-mobility shift assay and polymerase chain reaction. Sequencing and comparison of the 23 isolated clones revealed a 16-bp consensus sequence of 5'-GTG$\b{TGGC}$AGANCCNGG-3' in the light-strand (underlined residues were absolutely conserved), which nicely matched the genomic 13-bp terminator sequence 5'-$\b{TGGC}$AGAGCCCGG-3'. Moreover, mTERF binding assays of heteroduplex and single-stranded DNAs showed mTERF recognized the light strand in preference to the heavy strand. The preferential binding of mTERF with the light-strand may explain its distinct orientation-dependent termination activity.

Ribosomally Synthesiszed Antimicrobial Peptides (Bacteriocins) in Lactic Acid Bacteria: A Review

  • Nes, Ingolf F.;Yoon, Sung-Sik;Diep, Dzung B.
    • Food Science and Biotechnology
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    • v.16 no.5
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    • pp.675-690
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    • 2007
  • Bacteriocins in Gram-positive bacteria have attracted much attention because many have a strong antimicrobial activity also against bacteria outside the genera of the producers. Lantibiotics and the pediocin-like bactericins have attracted most attention since they kill a broad spectrum of Gram-positive bacteria including important pathogens. But many other promising Gram-positive bacteriocins have been thoroughly characterized. Recent studies have shown that bacteriocins may playa role in the intestinal flora to protect us against the food-borne pathogens. Bacterial genome sequencing has demonstrated that there may be an arsenal of such compounds and we are only seeing the top of the iceberg. The present review gives a short outlook of the field of bacteriocins with focus on lactic acid bacteria and includes recent findings.

Expression of Carboxypeptidase Z cDNA from Absidia zychae in saccharomyces cerevisiae and its characteristics (Saccharomyces cerevisiae에서 Absidia zychae 의 Carboxypeptidase A cDNA 의 발현과 특성)

  • 이병로;김종화
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.150-155
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    • 1995
  • Carboxypeptidase Z(CPZ) cDNA of Absidia zychae was experssed in Saccharomyces cerevisiae. The expressed CPZ(YCPZ) was secreted about 30 mg/l into the medium and has a little higher molecular weight than the wild type CPZ in SDS-PAGE. By the result of N-terminal amino acid sequencing, YCPZ has additional 15 amino acids residues in N-terminus of CPZ. But YCPZ shows no difference with CPZ in enzyme activity and substrate specificity. For the identification of processing mechanism of YCPZ, 36-Arg was changed to 36-Thr by site specific mutagenesis. Mutant YCPZ does not processed at 36-Thr. It was, therefore, concluded that the YCPZ was processed by KEX2. According to endo F treatment, high amount of carbohydrate was N-glycosylated in YCPZ.

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Removal Characteristics of Cyclic Ethers in Biological Wastewater Treatment System (고리형 에테르의 생물학적 처리 특성)

  • Lee, Sung-Ryul;Jeong, Yeon-Koo
    • Journal of Environmental Science International
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    • v.17 no.3
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    • pp.343-350
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    • 2008
  • The fate of two cyclic ethers, THF(Tetrahydrofuran) and 1,4-Dioxane, in conventional biological wastewater treatment plants was investigated using sequential activated sludge process. Removal efficiency of THF were about 86% in average, which was greater than that of 1,4-Dioxane, 30%. However, it was not clear whether the removal of cyclic ethers in biological system was caused by microbial activity or not. Thus treatability tests were conducted by batch experiments. The effects of mixing, aeration and the addition of activated sludge on the removal of cyclic ethers were investigated in batch experiments. THF was totally removed by mixing and aeration in 24 hours while removal ratio of 1,4-Dioxane was at most 30% for the same period. This results could be ascribed to the differences in Henry's law constants between the two chemicals. In addition, biological degradation including biosorption was not obviously observed in these batch tests.

Method for Cloning Biosynthetic Genes of Secondary Metabolites Including Deoxysugar from Actinomycetes

  • Sohng, Jae-Kyung;Oh, Tae-Jin;Kim, Chun-Gyu
    • BMB Reports
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    • v.31 no.5
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    • pp.475-483
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    • 1998
  • Many antibiotics contain partially deoxygenated sugar components that are usually essential for biological activity, affinity, structural stability, and solubility of antibiotics. Gene probes of the biosynthetic genes related with the deoxysugar were obtained from PCR. Primers were designed from the conserved peptide sequences of the known dTDP-D-glucose 4,6-dehydratases, which are the key step enzymes in the biosynthesis of deoxysugar. The primers were applied to amplify parts of dehydratase genes to 27 actinomycetes that produce the metabolites containing deoxysugar as structural constituents. About 180 and 340 bp DNA fragments from all of the actinomycetes were produced by PCR and analyzed by Southern blot and DNA sequencing. The PCR products were used as gene probes to clone the biosynthetic gene clusters for the antibiotic mithramycin, rubradirin, spectinomycin, and elaiophyrin. This method should allow for detecting of the biosynthetic gene clusters of a vast array of secondary metabolites isolated from actinomycetes because of the widespread existence of deoxysugar constituents in secondary metabolites.

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Effects of Experimental Drought on Soil Bacterial Community in a Larix Kaempferi Stand

  • Kim, Beomjeong;Choi, Byoungkoo
    • Journal of Forest and Environmental Science
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    • v.34 no.3
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    • pp.258-261
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    • 2018
  • Drought alters soil microorganisms; however, it is still not clear how soil microbes respond to severe drought conditions. In this study, the responses of soil bacterial community to experimental drought in a coniferous stand were examined. Six $6m{\times}6m$ plots with three replicates of control and drought treatments were delimited. PCR amplification and Illumina sequencing were conducted for cluster analysis of soil bacterial community and species richness and species diversity was analyzed. Along the 393 days of simulated drought from July 2016 to October 2017, soil bacterial species diversity slightly increased whereas species richness decreased in both control and roof plots. Moreover, soil bacterial species richness more decreased in roof plots than in controls. Combining these results, soil bacterial activity might have been altered by simulated drought.

Isolation and Characterization of Lactobacillus buchneri Strains with High ${\gamma}$-Aminobutyric Acid Producing Capacity from Naturally Aged Cheese

  • Park, Ki-Bum;Oh, Suk-Heung
    • Food Science and Biotechnology
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    • v.15 no.1
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    • pp.86-90
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    • 2006
  • Two lactic acid bacteria (LAB) with high ${\gamma}$-aminobutyric acid (GABA)-producing capacity were isolated from naturally aged cheese. Examination of the biochemical features using an API kit indicated that the two strains belonged to Lactobacillus. They were gram positive, rod-type bacteria, and fermented arabinose, melezitose, melibiose and xylose, but did not utilize cellobiose or trehalose. 16S rDNA sequencing analysis confirmed that they were Lactobacillus buchneri and Lactobacillus sp. They were accordingly named as Lactobacillus buchneri OPM-1 and Lactobacillus sp. OPM-2, and could produce GABA from MRS broth supplemented with 10 g/L of monosodium glutamate (MSG) at a productivity of 91.7 and 116.7 mg/L/hr, respectively. Cell extracts of L. buchneri OPM-1 and Lactobacillus sp. OPM-2 showed glutamate decarboxylase (GAD) activity, for which the optimum pH and temperature were 5.5 and $30^{\circ}C$, respectively.