• Title/Summary/Keyword: Molecular characterization

Search Result 3,085, Processing Time 0.025 seconds

The Characterization of Spherical Particles in S/G Sludge (S/G 슬러지 중 구형입자의 특성측정)

  • Pyo Hyung-Yeal;Park Yang-Soon;Park Sun-Dal;Park Yong-Joon;Park Kyoung-Kyun
    • Proceedings of the Korean Radioactive Waste Society Conference
    • /
    • 2005.06a
    • /
    • pp.129-136
    • /
    • 2005
  • There should not be ion exchange resin particles in S/G sludge. The suspicious spherical resin particles observed in S/G sludge sample were characterized for particle size distribution under optical microscope using the micro-technique, for element analysis by the electron probe micro analysis (EPMA), and for molecular identification by the IR spectroscopy The particle sizes are distributed from 1 to 200 ${\mu}m$ for the sludge, while 40 to 500 ${\mu}m$ for the spherical resin particles. The results of the elemental analysis showed different major impurities: Si, Al, Mn, Cr, Ni, Zn and Ti for the sludge particles, while Si, Cu, Zn for the spherical resin particles. However, both particles contain Fe as a matrix of hematite ($Fe_{3}O_4$). IR spectrum of the spherical particles was quite different from that of ion exchange resins used in S/G system. These results indicate that the spherical particles are not related to ion exchange resin particles and formed by the process of the sludge formation.

  • PDF

Nucleotide Sequence and Cloning of sfs4, One of the Genes Involved in the CRP-Dependent Expression of E. coli mal Genes. (CRP 의존성 maltose 대사 촉진 유전자 sfs4의 클로닝 및 염기배열 결정)

  • Chung, Soo-Yeol;Cho, Moo-Je;Jeong, Hee-Tae;Choi, Yong-Lark
    • Applied Biological Chemistry
    • /
    • v.38 no.2
    • /
    • pp.111-117
    • /
    • 1995
  • In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

  • PDF

Isolation and Characterization of Colletotrichum Isolates Causing Anthracnose of Japanese Plum Fruit (자두 탄저병균의 분리 및 동정)

  • Lee, Yong-Se;Ha, Da-Hee;Lee, Tae-Yi;Park, Min-Jung;Chung, Jong-Bae;Jeong, Byeong-Ryong
    • Korean Journal of Environmental Agriculture
    • /
    • v.36 no.4
    • /
    • pp.299-305
    • /
    • 2017
  • BACKGROUND: Although the filamentous fungal pathogen Colletotrichum species causing anthracnose disease on various fruits including peach, apple, persimmon and grape, there is no report on Japanese plum in Korea. METHODS AND RESULTS: In 2016, diseased fruits showing typical anthracnose symptoms of Japanese plum were collected in market and ochards. Diseased tissue was cut off and disinfected subsequently with 70% ethanol for 1 min, and in 1% sodium hypochloride solution for 1 min, followed by three washes with sterile distilled water. The disinfected tissues were placed onto potato dextrose agar (PDA), and incubated at $25^{\circ}C$ in the dark for 5 to 7 days. For single-spore isolation, conidia were scraped off the plate using a loop, and suspended with 10 mL sterile distilled water. One hundred microliter of the conidial suspension was spread on PDA plates and incubated at $25^{\circ}C$. Finally, one germinated conidium was transferred onto PDA plates. Morphological and cultural characteries of colonies and spores of isolated Colletotrichum were observed after 7 to 10 days incubation on PDA. Molecular identification of isolates were analyzed by comparing rDNA-ITS gene sequences with NCBI GeneBank. CONCLUSION: Of eleven isolates of Colletotrichum isolated from anthracnose diseased Japanese plum fruits, six were identified as C. acutatum, and five as C. gloeosporioides based on diagnostic characteristics such as colony growth rate, shape and size of conidia, and rDNA-ITS sequences. This is the first report of Colletotrichum causing the anthracnose on Japanese plum in Korea.

Purification and Characterization of Myrosinase in Dolsan Leaf Mustard(Brassica juncea) and Changes in Myrosinase Activity during Fermentation of Leaf Mustard Kimchi (돌산갓의 Myrosinase 분리 정제 및 갓김치 숙성 중 Myrosinase 활성도의 변화)

  • Park, Jeong-Ro;Park, Seok-Kyu;Cho, Young-Sook;Chun, Soon-Sil
    • Journal of the Korean Society of Food Culture
    • /
    • v.9 no.2
    • /
    • pp.137-142
    • /
    • 1994
  • Myrosinase in leaf mustard was purified and characterized to furnish a grounding information for utilizing the pungent taste and the potential antimicrobial capability of Dolsan leaf mustard to enhance the taste and storage life of kimchi. When myrosinase was purified from leaf mustard through a series of DEAE Sephadex, chromatofocusing and Con A Sepharose column chromatography, specific activity of the enzyme increased 7107-fold compared with that of crude enzyme preparation, and 18.8% yield was obtained. The purified myrosinase showed the optimum pH of 5.9, isoelectric point of 4.6, molecular weight of 129 kD, Km of 0.206 mM, and Vmax of $2.039\;{\mu}M{\cdot}min^{-1}{\cdot}mg\;protein^{-1}$, respectively. The optimum concentration of L-ascorbic for the maximum activity of the enzyme was 0.6 mM, and the enzyme activity decreased at a higher concentration of L-ascorbic acid than 0.6 mM, showing almost no enzyme activity at a L-ascorbic acid concentration of higher than 2.0 mM. Myrosinase activity in leaf mustard kimchi immediately after the kimchi was formulated was shown to be about 70 nmol/min/mg protein which decreased rapidly after 3 days of storage at $20^{\circ}C$, showing that less than half and almost none of the enzyme activity was retained in 4 and 10 days of storage, respectively.

  • PDF

Prevalence and Molecular Characterization of Tetracycline Resistance in $Enterococcus$ Isolates from Raw Milk Samples in Korea (원유시료에서 분리한 장구균 속 세균의 tetracycline 내성 유전자형 분석)

  • Kim, Ji-Hoon;Choi, Sung-Sook
    • Journal of Food Hygiene and Safety
    • /
    • v.27 no.1
    • /
    • pp.63-67
    • /
    • 2012
  • Antibiotic resistance in animal isolates of enterococci is a public health concern, because of the risk of transmission of antibiotic-resistant strains or resistance genes to humans through the food chain. This study investigated phenotypic and genotypic resistances profile of tetracycline in 245 $Enterococcus$ isolates from bovine milk. A total of 245 enterococci were isolated from 950 milk samples. The predominant strain was $E.$ $faecalis$ (n = 199, 81.2%) and $E.$ $faecium$ (n = 25, 10.2%). $E.$ $avium$ (n = 7, 2.9%), $E.$ $durans$ (n = 6, 2.5%), $E.$ $gallinarum$ (n = 4, 1.6%), and $E.$ $raffinosus$ (n = 4, 1.6%) were also isolated. Of the 245 enterococcal isolates 76.3% (n = 187) displayed tetracycline resistance (${\geq}16{\mu}g/ml$). Of the 187 tetracycline-resistant isolates, 83.4% (n = 156), 16.1% (n = 30), and 26.7% (n = 50) possessed the genes $tet$(M), $tet$(L), $tet$(S) respectively. While 3.2% (n = 6) of the tetracycline-resistant isolates possessed all three genes $tet$(M) + $tet$(L) + $tet$(S), 8.6% (n = 16), 16.0% (n = 30), and 2.7% (n = 5) of them possessed two genes $tet$(M) + $tet$(L), $tet$(M) + $tet$(S), and $tet$(L) + $tet$(S) respectively. The tetracycline resistance pattern investigated in this study was attributable mainly to the presence of $tet$(M).

Clinical Features, Response to Treatment, Prognosis, and Molecular Characterization in Korean Patients with Inherited Urea Cycle Defects

  • Yoo, Han-Wook;Kim, Gu-Hwan;Seo, Eul-Ju
    • Journal of The Korean Society of Inherited Metabolic disease
    • /
    • v.2 no.1
    • /
    • pp.77-79
    • /
    • 2002
  • The urea cycle, consisting of a series of six enzymatic reactions, plays key roles to prevent the accumulation of toxic nitrogenous compound and synthesize arginine de novo. Five well characterized diseases have been described, resulting from an enzymatic defect in the biosynthesis of one of the normally expressed enzyme. This presentation will focus on two representative diseases; ornithine transcarbamylase(OTC) deficiency and citrullinemia(argininosuccinate synthetase deficiency). OTC deficiency is one of the most common inborn error of urea cycle, which is inherited in X-linked manner. We identified 17 different mutations in 20 unrelated Korean patients with OTC deficiency; L9X, R26P, R26X, T44I, R92X, G100R, R141Q, G195R, M205T, H214Y, D249G, R277W, F281S, 853 del C, R320X, V323M and 10 bp del at nt. 796-805. These mutations occur at well conserved nucleotide sequences across species or CpG hot spot. The L9X and R26X lead to the disruption of leader sequences, required for directing mitochondrial localization of the OTC precursor. Their phenotypes are severe, and neonatal onset. The G100R, R277W and V323M mutations were uniquely identified in patients with late onset OTC deficiency. The other genotypes are associated with neonatal onset. Out of 20 patients with OTC deficiency, only 6 patients are alive; two were liver transplanted, and normal in growth and development at 2, 4 years after transplantation respectively. Citrullinemia is an autosomal recessive disease, caused by the mutations in the argininosuccinate synthetase(ASS) gene. We identified in 3 major mutations in 11 unrelated Korean patients with citrullinemia; G324S, $IVS6^{-2}$ A to G, and 67 bp ins at nt 1125-1126. Among these, the 67 base pair insertion mutation is novel. The allele frequency of each mutation is; G324S(45%), IVS6-2 A to G(32%), and 67 base pair insertion(14%). All patients are diagnosed at neonatal or infantile age. Interestingly, two patients presented with stroke like episode. Out of 11 patients, 5 patients died. Among 6 patients alive, one patient was successfully liver transplanted.

  • PDF

Characterization of a Fibrinolytic Enzyme from Pickled Anchovy (멸치젓갈유래의 혈전용해호소에 대한 특성)

  • Yang Woong-Suk;Lim Hak-Seob;Chung Kyung Tae;Kim Young-Hee;Huh Man Kyu;Choi Byung Tae;Choi Yung Hyun;Jeong Yong Kee
    • Journal of Life Science
    • /
    • v.15 no.3 s.70
    • /
    • pp.434-438
    • /
    • 2005
  • In the previous study, we isolated a myulchikinase (MK), which has fibrinolytic activity and cytotoxicity to the tumor cell line, from myl- chi-jeot-gal. In this study, the effect of NaCl concentration, metallic ions, pH, temperature, and plasminogen on the activity of MK was analysed. The MK activity was maintained at least $80\%$ activity up to $30\%$ NaCl, which indicates that the enzyme may be halotolerant. The optimal pH and temperature were 8 and $40^{\circ}C$, respectively. The fibrinolytic activity of MK was completely inhibited with 0.5 mM $Hg^{2+}$ and inhibited to $50^{\circ}C$ with 1 mM $Cu^{2+}\;and\;Zn^{2+}$. The MK showed strong activity in plasminogen- rich fibrin plate but not in plasminogen-free fibrin plate. The result indicates that the MK may be a plasminogen activator type fibrinolytic enzyme.

Characterization of Bacteriocin, lacticin YH-10, Produced by Lactococcus lactis subsp. lactis YH-10 Isolated from Kimchi (김치유산균인 Lactococcus lactis subsp. lactis YH-10가 생산하는 박테리오신의 특성)

  • Park, Eun-Min;Kim, Young-Hwa;Park, So-Jin;Kim, Yun-Im;Ha, Yu-Mi;Kim, Sung-Koo
    • Journal of Life Science
    • /
    • v.14 no.4
    • /
    • pp.683-688
    • /
    • 2004
  • A bacteriocin-producing lactic acid bacteria was isolated from Kimchi on MRS selective media with the use of Lactobacillus delbrueckii subsp. delbrueckii as an indicator strain. The strain YH-10 was identified as Lactococcus lactis subsp. lactis through the API test. The crude bacteriocin (freeze-dried 50% ammonium sulfate precipitate of culture supernatant) produced by the strain was named as lacticin YH-10. Lacticin YH-10 showed the growth inhibitory activity against Gram positive pathogenic bacteria and other lactic acid bacteria. The bacteriocin was inactivated by proteases such as protamex and aroase AP-10 and partially inactivated by amylase, proteinase K, trypsin, and papain. The lacticin YH-10 remained its activity with the treatment of heat at 10$0^{\circ}C$ for 60 min or the changes of pH 2 to 11. However, the activity was lost at high pH combined with the exposure to 10$0^{\circ}C$. The bacteriocin production of the strain was started in the exponential phase and stopped in the stationary phase. The approximate molecular mass of the bacteriocin produced by the strain was approximate 14 kDa in the analysis on SDS-PAGE.

Purification and Characterization of Exo-xylanase from Escherichia coli Cells Harboring the Recombinant Plasmid pMGl (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus Exo-xylanase의 정제 및 특성)

  • 문애란;최용진
    • Microbiology and Biotechnology Letters
    • /
    • v.20 no.5
    • /
    • pp.574-582
    • /
    • 1992
  • Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.

  • PDF

Molecular Cloning and mRNA Expression of the Hanwoo CAT/enhancer-binding Protein α(C/EBPα) Gene (한우 CCAAT/enhancer-binding protein α(C/EBPα) 유전자의 동정과 mRNA의 발현)

  • Jeoung, Y.H.;Lee, S.M.;Park, H.Y.;Yoon, D.H.;Moon, S.J.;Chung, E.R.;Kang, M.J.
    • Journal of Animal Science and Technology
    • /
    • v.46 no.6
    • /
    • pp.909-916
    • /
    • 2004
  • CCAAT/enhancer binding proteins(C/EBP) are a group of transcription factors expressed during preadipocyte differentiation. In the C/EBPs, C/EBPa plays an important role in lipid deposition and adipocyte differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo CIEBP$\alpha$ The Hanwoo C/EBP$\alpha$DNA includes a 1059 bp open reading frame encoding a protein of 353 amino acids. The CIEBPa amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The distribution of C/EBP$\alpha$ mRNA in various tissues of Hanwoo aged 12 months were investigated using Northern blotting analysis. The highest expression was detected in adipose tissue and more lower expression was detected in colon and lung. We also identified expression of C/EBPa mRNA in Hanwoo sirloin and adipose tissue aged 12, 26, and 30 months by real-time RT-PCR. The higest expression were detected at 26 months in the sirloin and at 12 and 26 months in the adipose tissue.