• Title/Summary/Keyword: ST purification

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Purification and Characterization of Chitinase from Antagonistic Bacteria Pseudomonas sp. 3098. (생물방제균 Pseudomonas sp. 3098이 생산하는 Chitinase의 정제 및 특성)

  • 이종태;김동환;도재호;김상달
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.515-522
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    • 1998
  • Plant root rotting fungi, Fusarium solani are suppressed their growth by the chitinase which is produced from the antagonistic soil bacteria. The chitinase producable antagonistic bacterium Pseudomonas sp. 3098 was selected as a powerful biocontrol agent of F. solani from ginseng rhizosphere. The antagonistic Pseudomonas sp. 3098 was able to produce a large amount of extracellular chitinase which is key enzyme in the decomposition of fusarial hypal walls. The chitinase was purified from cultural filtrate of Pseudomonas sp. 3098 by the procedure of ammonium sulfate precipitation, anion exchange chromatography, gel filtration on Bio-Gel P-100, and 1st and 2nd hydroxyapatite chromatography. The molecular mass of the purified enzyme was ca. 45 kDa on SDS-FAGE. The optimal pH and temperature for the activity of purified chitinase were 5.0 and 45$^{\circ}C$, respectively. The enzyme was stable in pH range of 5.0 to 9.0 up to 5$0^{\circ}C$ The enzyme was significantly inhibited by metal compounds such as FeCl$_2$, AgNO$_3$ and HgCl$_2$, and was slightly inhibited by p-CMB, iodoacetic acid, urea, 2,4-DNP and EDTA. The enzyme had ability of digestion on colloidal chitin and chitin from shrimp shell, but could not digest chitosan and chitin from crab shell. Km value of the enzyme was 0.11% on colloidal chitin, and the maximum hydrolysis rate of the enzyme was 34% on colloidal chitin.

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Treatment Efficiency of Pollutants in Constructed Wetlands under Different Hydroponic Wastewater Injection Methods and Characteristic of Filter Media (시설하우스 폐양액 주입방법 및 여재 특성에 따른 인공습지에서 수질오염물질의 처리효율)

  • Seo, Dong-Cheol;Park, Jong-Hwan;Cheon, Yeong-Seok;Park, Seong-Kyu;Kim, Ah-Reum;Lee, Won-Gyu;Lee, Sang-Won;Lee, Seong-Tae;Cho, Ju-Sik;Heo, Jong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.29 no.2
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    • pp.146-151
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    • 2010
  • In order to improve T-N and T-P removal in HF (horizontal flow)-HF hybrid constructed wetlands by natural purification method for treating the hydroponic wastewater in greenhouses, the efficiency of water treatment as affected by the injection method of hydroponic wastewater, the addition of special filter media, the particle size of filter media, and the injection ratio of hydroponic wastewater in $1^{st}$ HF and $2^{nd}$ HF beds were investigated in small-scale HF-HF hydroponic wastewater treatment apparatus. Removal rate of T-P in the water in HF-HF hydroponic wastewater treatment apparatus with calcite as affected by addition method of special filter media was higher than that in HF-HF hydroponic wastewater treatment apparatus with other filter media. Removal rate of BOD, COD, SS, T-N and T-P in the water in mixed filter media with calcite were 86, 84, 87, 50 and 97%, respectively. Removals of pollutants except for T-P in the water were slightly different. Therefore, it should be considered that the removal rate of T-P was good for calcite in HF-HF hydroponic wastewater treatment apparatus. To improve T-N and T-P removal, the optimum particle size of filter media was 1.2 mm, and the optimum injection ratio ($1^{st}$ HF bed : $2^{nd}$ HF bed) of hydroponic wastewater was 60:40.

Purification and Properties of an Extracellular Chitinase from Streptomyces sp. (Streptomyces속 균주로 부터 생산되는 Chitinase의 정제 및 그 성질)

  • Hong, Yong-Ki;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.7 no.3
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    • pp.149-155
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    • 1979
  • Streptomyces sp. 115-5 was selected as the most active microorganism of about 200 strains for the production of chitinase. The enzyme was purified by (NH$_4$)$_2$SO$_4$ treatment, 1st-Sephadex G-100, DEAE-Cellulose, 2nd-Sephadex G-100 column chromatography, and evidence for homogenity was obtained from CM-Sephadex C-50 column chromatography and polyacylamide gel electrophoresis. The purified enzyme hydrolyzed chitin (N-acetyl glucosamine polymer) and chitosan (glucosamine polymer) but not cellulose. And with chitin as the substrate, a Km value of 3.6 mg of chitin per ml and a Vmax of 100 $\mu$mo1e fer hr were found. The activation of the chitinase was 3.66 kcal per mole. The molecular weight of the enzyme was esti-mated about 56,000 daltons by Sephadex G-100 chromatography and isoelectric point as pH 3.0.

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Studies on the Inulin Hydrolyzing Enzyme from Aspergillu sp. (C-58) (III) - Purification of inulase (P-I) from Aspergillus sp. (C-58) - (Aspergillus sp. (C-58)균주가 생산하는 Inulin 분해효소에 관한 연구 - Aspergillus sp. C-58균주가 생산하는 inulase P-I의 정제 -)

  • Kwon, Tae-Jong;Seu, Jung-Hwu
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.47-52
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    • 1983
  • The extracellular inulase produced by Aspergillus sp. C-58 was isolated by pH and charcoal treatment, precipitation with ammonium sulfate from the crude extract, and separated into 3 fractions (P-I, II, III) by DEAE-cellulose column chromatography in the ratio of 31.1:1.7:1 with respect to the activity. The ratio of inulase activity to sucrase activity of P-I, P-II and P-III fraction was 0.23, 0.24 and 1.1 respectively. The enzyme P-I fraction was purified 482 fold with a 22.8% yield by DEAE-Sephadex A-50, Sephadex G-75, Sephadex G-100 (1st and 2nd) column chromatography, and appeared homogeneous on polyacrylamide disc gel electrophoresis and ultracentrifugation.

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Purification of Oxytocin-related Peptide, Isotocin from the Brain of Conger Eel Conger myriaster (붕장어(Conger myriaster)의 뇌로부터 Oxytocin-related Peptide, Isotocin의 정제)

  • GO Hye-Jin;KIM Chan-Hee;KIM Eun Jung;KIM In Hae;AN Sang Hyun;SOHN Hee-Young;PARK Jin-IL;PARK HEE Yun;YOON Ho Dong;PARK Nam Gyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.38 no.5
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    • pp.286-290
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    • 2005
  • Oxytocin (OT)-related peptide, isotocin was purified from the brain extract of conger eel (Conger myriaster) using reverse-phase, ion-exchange and size exclusion high performance liquid chromatography (HPLC). The sequence of the peptide, with a molecular weight of 967.30 Da, was determined as Cys-Tyr-Ile-Ser-Asn­Cys-Pro-Ile-Gly-$NH_2$, where the Cys between 1st and 6th residues made an intramolecular disulfide bridge by the automated amino acid sequence analysis and MALDI-TOF mass spectrometry. The sequence was confirmed by identical elution with the purified and synthetic peptide using the HPLC system. As a result of homology investigation, the primary structure of this peptide was the same as that of OT -superfamily member, isotocin. The synthetic peptide showed a contractile activity at a minimal effective concentration of $10^{-7}M$ on the intestinal smooth muscle of goldfish (Carassius auratus).

Fish Community Characteristics in Hwapocheon Wetland, Korea (화포천 습지의 어류군집 특성)

  • Ko, Myeong-Hun;Choi, Kwang-Seek;Lim, Jeong-Cheol
    • Korean Journal of Environment and Ecology
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    • v.36 no.2
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    • pp.165-176
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    • 2022
  • This study surveyed the characteristics of fish communities in Hwapocheon Wetland, Korea, from May to September 2020. The survey collected 735 objects in 21 species belonging to 7 families from 8 survey stations. The dominant and subdominant species were Hemiculter eigenmanni(23.8%) and Micropterus salmoides(10.3%), respectively. The next most abundant species were Zacco platypus(9.5%), Carassius auratus(9.4%), Pseudorasbora parva(9.0%), Squalidus chankaensis tsuchigae(6.7%), Acheilognathus macropterus(5.4%), Lepomis macrochirus(5.2%), Pseudogobio esocinus(4.1%), Opsariichthys uncirostris amurensis(3.7%), and Carassius cuvieri(3.3%). Among the fish species collected, one species, Culter brevicauda, was class II endangered wildlife designated by the Ministry of Environment, and one species,S. c. tsuchigae(4.8%), was endemic to Korea.Additionally, three exotic species (M. salmoides, L. macrochirus, and C. cuvieri) and one landlocked species (Rhinogobius brunneus) were collected. Compared to previous studies, the proportion of fish living in the running water area tended to decrease, the proportion of fish living in the water purification area tended to increase, and ecosystem-disturbing species (M. salmoides and L. macrochirus) tended to increase gradually. Results of fish community analysis showed that the mainstream stations (St. 1, 3, 4, 5, 6, and 8) had low dominance, but high diversity and richness, and other stations (St. 2 and 7) had high dominance but low diversity and richness. The river health (index of biological integrity) evaluated using fish was assessed as bad (6 stations), normal (1 station), and very bad (1 station). The water quality grade was assessed as slightly bad due to the chemical oxygen demand (COD), total organic content (TOC), suspended solid (SS), and total coliforms (TC). The annual water quality showed a gradually increasing trend of biological oxygen demand (BOD), COD, SS, and chlorophyll-a. The stable life of fish and the improvement of river health in Hwapocheon Wetland require water quality improvement and the systematic management of ecosystem-disturbing species (M. salmoidesand L. macrochirus).

The Purification and Immunogenicity of TB-14 Recombinant Protein of Mycobacterium tuberculosis (결핵균 특이 TB-14 재조합 단백질의 분리 및 세포성 면역반응에 미치는 영향)

  • Song, Ho-Yeon;Kim, Young-Hee;Kim, Chang-Hwan;Min, Young-Ki;Kim, Dae-Joong;Ko, Kwang-Kjune
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.3
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    • pp.239-247
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    • 2006
  • Background: Culture filtrate proteins secreted by mycobacteria are thought to play an important role in inducing protective immunity and to develop new methods for diagnosing tuberculosis. Methods: A culture filtrate protein of M. avium that was strongly reactive with goat antiserum against M. intracellulare was constructed. Its homologous protein (TB-14) in M. tuberculosis was cloned, expressed and purified. The inductions of IFN-${\gamma}$ stimulated with $10{\mu}g$ of TB-14 recombinant protein and $10{\mu}g$ PPD were estimated by using whole bloods from seven PPD (-) subjects, seven PPD (+) healthy volunteers and nine tuberculosis patients. Results: M. avium culture filtrate protein was confirmed as a hypothetical protein that was termed contig 116. A novel 14-kDa recombinant protein (TB-14) of M. tuberculosis was composed of 148 amino acids, including 30 amino acids of the signal peptide, and it showed 78% homology with M. avium. In the PPD (+) healthy volunteers, recombinant TB-14 protein strongly induced the secretion of IFN-${\gamma}$ in whole blood cultures. Conclusion: These results suggest that TB-14 recombinant protein might play an important role in inducing cell-mediated immunity against tuberculosis. Furthermore, TB-14 protein antigen and its antiserum will be available for the development of new diagnostic tools for tuberculosis.

Optimum Pre-treatment Method in Constructed Wetlands by Natural Purification Method for Treating Livestock Wastewater (자연정화공법에 의한 인공습지에서 효과적인 축산폐수처리를 위한 최적 전처리방법 구명)

  • Park, Jong-Hwan;Seo, Dong-Cheol;Kim, Ah-Reum;Kim, Sung-Hun;Lee, Seong-Tea;Jeong, Tae-Uk;Choi, Jeong-Ho;Lee, Sang-Won;Cho, Ju-Sik;Kim, Hyun-Ook;Heo, Jong-Soo
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.3
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    • pp.425-433
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    • 2011
  • In order to obtain optimum pre-treatment methods and improve T-N and T-P removal efficiencies, removal rates of pollutants in small-scale livestock wastewater treatment apparatus with water plant filtration bed or activated sludge tank were investigated. Based on the results from the optimum pre-treatment in small-scale livestock wastewater treatment apparatus, removal efficiencies of pollutants in livestock wastewater treatment plant with water plant filtration and activated sludge beds. The removal rates of COD, SS, T-N, and T-P in effluent were 83, 89, 63 and 87% in small-scale livestock wastewater treatment apparatus with water plant filtration bed, respectively. The removal rates of COD, SS, T-N, and T-P in effluent were 96, 95, 86 and 92% in small-scale livestock wastewater treatment apparatus with activated sludge tank, respectively. For increasing the COD, SS, T-N, and T-P removals in small-scale livestock wastewater treatment apparatus, the water plant filtration and activated sludge beds are recommended. In livestock wastewater treatment plant with water plant filtration ($1^{st}$ treatment) and activated sludge ($2^{nd}$ treatment) beds, the concentrations of COD, SS, T-N, and T-P in effluent were 39, 15, 42 and $1mg\;L^{-1}$, respectively. It was shown that the concentrations of COD, SS, T-N, and T-P met acceptable effluent quality standard for livestock wastewater. Based on the above results, the removal rates of COD, SS, T-N, and T-P in effluent were over 99.8, 99.9, 99.2, and 99.9% in livestock wastewater treatment plant, respectively.