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The Isolation of Bacillus sphaericus 366M-9 Producing New Cephalosporin-C Deacetylase (CAH) and its Enzymatic Characterization (신규 Cephalosporin-C Deacetylase(CAH) 생산 균주인 Bacillus sphaericus 366M-9의 선발 및 그 효소학적 특성)

  • 이승훈;권태종;이동희
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.224-229
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    • 2004
  • Several microorganisms (esterase-producing group) were isolated by the solid selective media containing-naphtylacetate. Among them, strain 366M-9 having a high activity of cephalosporin-C deacetylase (CAH; EC 3.1.1.41) was selected. The strain 366M-9 was identified as Bacillus sphaericus on the basis of morphological, physiological, and biochemical characteristics. The production of CAH reached at maximum value after 32 hrs, when cultivated in the optimal medium containing dextrin 2.5%, peptone 2.5%, sodium chloride 0.5%, dipotassium phosphate 0.25%, ferrous sulfate 0.02%, and 7-ACA 0.1% at $30^{\circ}C$ with initial pH 6.0. The CAH was purified by 3 steps with ammonium sulfate precipitation, adsorption chromatography on hydroxyapatite column, and Sephadex G-200 gel chromatography. The final enzyme preparation was homogeneous as judged by the analysis of SDS-PAGE and HPLC. Optimum temperature and pH for CAH activity were $50{\circ}C$ and around 7.0, respectively. And the enzyme was stable at pH 6.0~8.0, up to $50^{\circ}C$. The Michaelis-Menten constants ($K_{m}$ ), $V_{max}$ were 0.87 mM and 1.22 unit/ml, respectively.

Foliar Retention of the Herbicide Pyribenzoxim(1% EC), and Its Effects on Herbicidal Activity and Rice Phytotoxicity (Pyribenzoxim 1% 유제(乳劑)의 경엽(莖葉) 부착량(附着量)과 약효(藥效), 약해(藥害)의 관계(關係))

  • Koo, Suk-Jin;Kim, Jeong-Su;Lee, Jae-Hwan
    • Korean Journal of Weed Science
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    • v.18 no.4
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    • pp.304-313
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    • 1998
  • Foliar retention of pyribenzoxim (1% EC) was measured using the fluorescent dye rhodamine B, and related to efficacy and phytotoxicity to barnyardgarss (Echinochloa crusgalli) and rice (Oryza sativa cv. Chucheong), respectively. Effects of nozzle types (8002E flat-fan and disk-type), addition of adjuvant, variation of herbicide concentration or spray volume were compared. In barnyardgrass, foliar retention of pyribenzoxim at a recommended condition (application rate : 30g ai/ha, spray volume : 1000 L/ha) was 2.3 to 2.7 or 1.4 to $1.5{\mu}g$ ai/g fresh foliage when sprayed using the disk-type nozzle with or without adjuvant, respectively, and 0.6 to 0.7 or 0.3 to $0.5{\mu}g$ ai/g fresh foliage when sprayed using the flat-fan nozzle with or without adjuvant, respectively. The slope of increase in foliar retention was 1.0 to 1.8 when application rates increased from 10 to 60g ai/ha at 1000 L/ha, while that was 1.6 to 2.4 when spray volume increased from 330 to 2000 L/ha at $30{\mu}g$ ai/L concentration. Foliar retention of pyribenzoxim had a close relationship with herbicidal activity; regardless of spray conditions, retention to provide 90% control was about $0.8{\mu}g$ ai/g fresh foliage, and below this retention amount, efficacy decreased almost linearly. In rice, foliar retention at the recommended condition was 1.9 to 2.3 or 1.2 to $1.3{\mu}g$ ai/g fresh foliage when sprayed using the disk-type nozzle with or without adjuvant, respectively, and 0.6 to 0.9 or $0.3{\mu}g$ ai/g fresh foliage when sprayed using the flat-fan nozzle with or without adjuvant, respectively. The slope of increase in foliar retention was 1.0 to 2.8 when application rates increased from 30 to 120g ai/ha at 1000 L/ha, while that was 1.3 to 4.4 when spray volume increased from 1000 to 4000 L/ha at $30{\mu}g$ ai/L concentration. Despite the great difference in retention, rice phytotoxicity was not observed in any of these spray conditions, suggesting retention differences within 4-fold increase of application rate or spray volume do not affect rice safety. When pyribenzoxim 1EC was sprayed in tank-mix with several other commercial pesticide formulations, its retention to rice foliage tended to increase by 30 to 50%.

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Parthenogenetic Activation and Development of Freshly Matured Bovine IVM Oocytes (체외성숙 직후 소 난포란의 단위발생과 체외발육능)

  • 정희태;임석기;오세훈;박춘근;양부근;김정익
    • Korean Journal of Animal Reproduction
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    • v.21 no.1
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    • pp.71-78
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    • 1997
  • This study was conducted to investigate the activation condition of freshly matured bovine IVM oocytes for use as a cytoplasmic recipient in nuclear transfer. Bovine oocytes matured in vitro for 22-24 h were treated with various activation conditions. In Experiment 1 in vitro matured oocytes were treated with electric stimuIus (ES; 2 pulses of 1.25 kV/cm for 70 ${\mu}{\textrm{s}}$ec, each pulse 1 sec apart), ethanol (ET; 7%, 5min) , Ca$^2+$-ionophore(A23187; 10$\mu\textrm{g}$/ml, 5min) and cycloheximide(CH; 10$\mu\textrm{g}$/ml, 6 h). Activation rates were similar in treatments with ES, ET and A23187(48.8~54.3%), however, significantly reduced with CH treatment(15.9%, P

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Ecotoxic Evaluations of BDE-47 and BDE-209 using Rotifer (Brachionus plicatilis) (해산로티퍼 (Brachionus plicatilis)를 이용한 브롬화난연제 (BDE-47, BDE-209)의 생태독성평가)

  • Choi, Hoon;Lee, Ju-Wook;Park, Yun-Ho;Lee, In-Seok;Heo, Seung;Hwang, Un-Ki
    • Korean Journal of Environmental Biology
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    • v.36 no.1
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    • pp.43-49
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    • 2018
  • The toxic assessment of the PBDEs (BDE-47, BDE-209) has been comprehensively investigated by using the rates of survival and population growth in the marine rotifer, Brachionus plicatilis. Chiefly, the survival rate was determined after a measurement of 24 hours of exposure to the BDE-47 (2,2'4,4'-Tetrabromodiphenyl ether) and the BDE-209 (2,2',4,4'-Decabromodiphenyl ether) was performed. The BDE-47 reduced survival rate in dose-dependent manner and a significant reduction were noted to have occurred at a concentration of greater than $3.9mg\;L^{-1}$, but the BDE-209 had no effect which was subsequently observed in this study. The population growth rate (r) was determined after 72 hours of exposure to toxicants in the study. It was observed that the r value in the controls (absence PBDEs) was greater than 0.5, and that it decreased as the dose-dependent manner as recorded. The survival rate when exposed to BDE-47 and BDE-209, $EC_{50}$ value was $13mg\;L^{-1}$ and $>1,000mg\;L^{-1}$, and population growth rate was $3.67mg\;L^{-1}$ and $862.75mg\;L^{-1}$, respectively. Therefore, the BDE-47 is considered to be 76-235 times more harmful than the BDE-209 as noted. In this study, the ecotoxicological bioassay using a noted survival rate and population growth rate of B. plicatilis can be used as a baseline data for the continued establishment of the environmental quality standard of the incidences of the BDE-47 and BDE-209 in a marine environment.

Characteristics of Manure and Estimation of Nutrient and Pollutant of Holstein Dairy Cattle (홀스타인 젖소 분뇨의 특성과 비료성분 및 오염물질 부하량 추정)

  • Choi, D.Y.;Choi, H.L.;Kwag, J.H.;Kim, J.H.;Choi, H.C.;Kwon, D.J.;Kang, H.S.;Yang, C.B.;Ahn, H.K.
    • Journal of Animal Science and Technology
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    • v.49 no.1
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    • pp.137-146
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    • 2007
  • This study was conducted to determine fertilizer nutrient and pollutant production of Holstein dairy cattle by estimating manure characteristics. The moisture content of feces was 83.9% and 95.1% for urine. The pH of feces and urine were in the ranges of 7.0~7.4 and 7.5~7.8, respectively. The average BOD5, COD, SS, T-N, T-P concentrations of the dairy feces were 18,294, 52,765, 102,889, 2,575, and 457mg/ℓ, respectively. Dairy urine showed lower levels of BOD5(5,455mg/ℓ), COD(8,089mg/ℓ), SS(593mg/ℓ), T-N(3,401mg/l), and T-P(13mg/ℓ) than feces. The total daily produced pollutant amounts of a dairy cow were 924.1g(Milking cow), 538.8g(Dry cow), 284.4g(Heifer) of BOD5, 2,336.5g (Milking cow), 1,651.8g(Dry cow), 734.1g(Heifer) of COD and 4,210.1g(Milking cow), 2,417.1g(Dry cow), 1,629.1g(Heifer) of SS and 194.8g(Milking cow), 96.4g(Dry cow), 58.3g(Heifer) of T-N and 24.0g(Milking cow), 10.2g(Dry cow), 6.1g(Heifer) of T-P. The calculated amount of pollutants produced by a 450kg dairy cow for one year were 181.3kg of BOD5, 492.5kg of COD, 899.9kg of SS, 36.0kg of T-N and 4.1kg of T-P. The total yearly estimated pollutant production from all head(497,261) of dairy cattle in Korea is 90,149 tons of BOD5, 244,890 tons of COD, 447,491 tons of SS, 17,898 tons of T-N and 2,008 tons of T-P. The fertilizer nutrient concentrations of dairy feces was 0.26% N, 0.1% P2O5 and 0.14% K2O. Urine was found to contain 0.34% N, 0.003% of P2O5 and 0.31% K2O. The total daily fertilizer nutrients produced by dairy cattle were 197.4g (Milking cow), 97.4g(Dry cow), and 57.9g(Heifer) of Nitrogen, 54.2g(Milking cow), 22.2g(Dry cow), and 14.2g(Heifer) of P2O5 and 110.8g(Milking cow), 80.4g (Dry cow), and 39.5g(Heifer) of K2O. The total yearly estimated fertilizer nutrient produced by a 450kg dairy animal is 36.2kg of N, 8.8kg of P2O5, 24.6kg of K2O. The estimated yearly fertilizer nutrient production from all dairy cattle in Korea is 18,000 tons of N, 4,397 tons of P2O5, 12,206 tons of K2O. Dairy manure contains useful trace minerals for crops, such as CaO and MgO, which are contained in similar levels to commercial compost being sold in the domestic market. Concentrations of harmful trace minerals, such as As, Cd, Hg, Pb, Cr, Cu, Ni, Zn, met the Korea compost standard regulations, with some of these minerals being in undetected amounts.