• 한국식품영양과학회지
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• 제24권4호
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• pp.570-577
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• 1995

Aspergillus sp. SB-2704 was selected for its strong polygalacturonase activity among various strain of mold found in soil. It was found that production of polygalacturonase reached to maximum when the wheat bran medium containing 1% polypepton, 1% glucose, and 0.2% FeSO4 were cultured for 3 days at 35$^{\circ}C$. Polygalacturonase was purified 20.90 fold from Aspergillus SB-2704. The purification procedures include ammonium sulfate treatment, gel filtration on Sephdex G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 4.34%. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-polyacrylamide gel electrophoresis, the molecular weight was estimated to be 36,000. The optimum pH for the enzyme activity was 5.5 and optimum temperature was 5$0^{\circ}C$. The enzyme is stable in acidic condition. The activity of purified enzyme was inhibited by Pb2+, Hg2+ and Ba2+, whereas activated by Cu2+, Mn2+, Mg2+ and Fe2+. The activity of polygalacturonase was inhibited by the treament wit maleic anhydride, iodine, and EDTA. The result indicate the possible involvement of histidine and metal ion at active site.

• 한국식물병리학회지
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• 제10권1호
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• pp.13-17
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• 1994

A semiselective agar medium (XCO) was developed for the isolation of bacterial blight pathogen, Xanthomonas campestris pv. oryzae, from rice seed. The medium contained yeast extract 1.0 g, peptone 2.0 g, sucrose 5.0 g, sodium glutamate 1.0 g, FeSO4.7H2O 0.05 g, Fe.EDTA 1 mg, cephalexin 20 mg, Evan blue (0.1%) 1.5 ml, bromcresol purple (0.1%) 2.5ml, cycloheximide 100 mg and agar 15.0 g per liter. Colonies of X. c. pv. oryzae were 4~5 mm in diameter, smooth, round, blue (darker center) and convex after 6 days incubation at 28$^{\circ}C$. The recovery of 6 isolates of X. c. pv. oryzae on the XOC medium ranged from 81% to 120% (mean 98.2%) in comparison to Wakimoto's medium. The number of saprophytic bacteria from 10 rice seed lots on XCO medium was reduced to 70.4% of that on Wakimoto's medium. The recovery of X. c. pv. oryzae added to rice seed on XOC medium ranged from 67% to 87% (mean 75.6%) of that on Wakimoto's medium.

• 식품기술
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• 제11권4호
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• pp.49-60
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• 1998

Ascorbate-$Fe^3+$-EDTA에 의해 생성된 hydroxyl radical (HO.)을 이용하여 단백질과 지질의 산화적 손상을 유도하였으며, 이 손상에 대한 멜라토닌(melatonin), glutathione, $\alpha$-tocopherol의 방어효과를 분석하였다. 단백질로는 bovine serum albumin (BSA), 지질로는 phosphatidyl-choline (PC) liposome을 이용하였다. 그리고 BSA와 PC liposome의 혼합계에서도 각 항산화제의 효능을 분석하였다. 단백질의 산화적 손상은 carbonyl기의 생성 및 단백질 절단을, 지질산화는 lipid peroxidation (LPO, malondialdehyde 와 4-hydroxyalkenal) 분석을 통하여 조사하였다. 단백질의 산화는 멜라토닌과 glutathione이, 지질산화는 멜라토닌과 $\alpha$-tocopherol이 효과적으로 억제하였다. 그러나 glutathione은 지질, $\alpha$-tocopherol은 단백질의 산화를 억제하지 못하였다. BSA와 PC liposome이 동시에 존재하는 혼합물에서는 수용성과 지용성 특성을 모두 지닌 멜라토닌은 단백질, 지질 모두에 대해 산화방지효과가 있으나 수용성인 glutathione은 단백질에, 지용성인 $\alpha$-tocopherol은 지질에만 주로 효과가 있는 것으로 조사되었다.

• KSBB Journal
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• 제15권3호
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• pp.318-322
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• 2000

An oxygen-sensitive fumarate reductase has been purified from the cytosol fraction of the Enterococcus faecalis RKY1 grown anaerobically on a defined medium containing glycerol and fumarate. A major portion of the purification was performed with employing Triton X-100 and reducing agents by Phenyl-sepharose CL-4B DEAE-sepharose and Dephadex G-150 The final activity was 0.42 unit/mg. The deduced molecular mass of active band was 66 kDa. The optimal pH and temperature for the activity were 7.0 and 38$^{\circ}C$ respectively. The enzyme activity was not affected by 1mM metal ions such as bacl2 $.$2H2O HgCl2 MnCl2$.$4H2O ZnCl2 CuCl2$.$2H2O Mgcl2$.$6H2O FeSo4$.$7H2O and by EDTA. Partially purified enzyme ws yellow in color ; spectroscopic study indicated the presence of flavins as a cofactor.

• Mycobiology
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• 제32권2호
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• pp.74-78
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• 2004

An alkaline protease produced by Aspergillus niger C-15 was purified and characterized. The enzyme was purified 19.41-fold with a specific activity of 74150 U/mg and a recovery of 34.4% by gel filtration and ion exchange chromatography. The molecular weight of the enzyme was estimated to be 34 kDa. The optimum pH and temperature for the protease activity were pH 8.0 and $60^{\circ}C$, respectively. The enzyme activity inhibited by EDTA suggests that the preparation contains a metalloprotease. The enzyme activity of the metalloprotease was completely inhibited by 5 mM $HgCl_2$ and $FeCl_3$, while partially inhibited by $CuSO_4$, and $MnCl_2$. When polyols such as glycerol, mannitol, sorbitol and xylitol, were added to the reaction medium, most polyols tested enhanced protease activity. Especially, glycerol showed the highest effect. The alkaline metalloprotease was stable at high temperature and retained more than 90% of the initial activity at $60^{\circ}C$ and 86.4% under addition of glycerol.

• Journal of Plant Biology
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• 제31권4호
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• pp.309-315
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• 1988

Leaf mesophy11 protoplast cultures from six Nicotiana species, N. debneyi, N. rustica, N. amplexicaulis, N. glauca, N. glutinosa, and N. sylvestris were carried out. When we reduced the NH4NO3 and Fe.EDTA concentration to 1/3(7 mM) and 1/10(10$\mu$M) from the Murashige and Skoog medium respectively, cell division of the protoplasts was efficiently induced in four Nicotiana species, N. debneyi, N. rustica, N. amplexicaulis and N. glauca. However, other two species, N. glutinosa and N. sylvestris were failed in inducing cell division at the same culture condition. The protoclone calluses derived from four Nicotiana species were consequently regenerated on a MS basal medium supplemented with the appropriate auxin and cytokinin.

• 한국환경성돌연변이발암원학회지
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• 제25권2호
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• pp.71-75
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• 2005

Green tea and their major constituents such as catechins are famous materials for their anti-oxidative and anti-carcinogenic activity, but many compounds with reducing power can promote the oxidation in their oxidized form or in the presence of metal ion. We investigated the pro-oxidative effect of the ethanol extract equivalent up to 30mg of dried weight of green tea leaves in four in vitro systems which could be used for detecting DNA damage. Although ethanol extract of green tea did not show significant mutagenicity in Salmonella typhimurium TA102, which is sensitive strain to oxidative stress, it degraded deoxyribose extensively in the presence of $FeCl_3-EDTA$ complex, promoted 8-oxoguanine formation in the live bacteria cell, Salmonella typhimurium TAI04, and cleaved super coiled DNA strand with the help of copper ion. It suggested that green tea, famous anti-oxidative material, can be pro-oxidant according to the condition of extraction or metal existence.

• 미생물학회지
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• 제30권4호
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• pp.299-304
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• 1992

An intracellular protease form cells of Pseudomonas carboxydovorans DSM 1227 grown on carbon monoxide was purified 57-fold in six steps to homogeneity with a yield of 4.3% using azocoll as a substrate. The molecular weight of the enzyme was determined to be 150,000. Sodium dodecyl sulfate-gel electrophoresis revealed the purified enzyme to be a dimer with two identical subunits of molecular weight 72,000. The enzyme was stimulated by $Mg^{2+}$ but was inhibited completely by $Cd^{2+}$ $Fe^{2+}$ $Hg^{2+}$, and $^Zn{2+}$ The enzyme activity was also inhibited by EDTA, EGTA, phenylmethylsulfonyl fluoride, and phenyl glyoxal, but was increased by 1-ethyl-3(dimethyl aminopropyl fluoride, and phenyl glyoxal, but was increased by 1-ethyl-3(dimethyl aminopropyl)carbodiimide, iodoacetamide and dithiothereitol. The optimal pH and temperature for the enzyme reaction were found to be 7-8 and 50.deg.C, respectively. Casein and bovine serum albumin were hydrolyzed by the enzyme, but carbon monoxide dehydrogenase was not.

• Bulletin of the Korean Chemical Society
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• 제26권6호
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• pp.943-946
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• 2005

An effective spectrophotometric determination of palladium with 4-(2-pyridylazo)-resorcinol (PAR) using molten naphthalene as a diluent has been studied. A red complex of palladium with PAR is formed at 90 ${^{\circ}C}$. In the range of pH 9.0-11.0, the complex is quantitatively extracted into molten naphthalene. The organic phase is anhydrously dissolved in $CHCl_3$ to be determined spectrophotometrically at 520 nm against the reagent blank. Beer’s law is obeyed over the concentration range of 0.1-2 ${\mu}g{\cdot}mL^{-1}$. The molar absorptivity and Sandell’s sensitivity are 8.0 ${\times}\;10^5\;L{\cdot}mol^{-1}{\cdot}cm^{-}1\;and\;0.49\;{\mu}g{\cdot}cm^{-2}$ respectively. From the results of tolerance limits, it was found that there was no interferences were observed for most of the ions examined and those somewhat high interferences by Co(II), Fe(II) and Bi(III) could be effectively masked by EDTA.

• Journal of Microbiology
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• 제33권4호
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• pp.283-288
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• 1995

We have identified hemolysin rendering virulency of Vibrio anguilarum grown at 23.deg.C which was evaluated on human RBCs. Hemolysin itself was separated as a single band on non-denaturing gel electrophoresis. Vibrio hemolysin was destroyed by trypsin and proteinase K and was heat labile. Optimal pH for activity was around pH 6 while pI of the molecule was recognized as 5.7, with relative distance (R$\sub$f/) on non-denaturing gel was 0.7. Addition of EDTA and FeCI$\sub$3/ drew the possibility that the production of hemolysin was mainly induced to overcome iron deficiency inside host animalsd infection.