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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.

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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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.