• Journal of Environmental Science International
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• v.7 no.6
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• pp.853-857
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• 1998

Purification of the isocitrate lyase extracted from Microbacterium laevaniformans was investigated. The isocitrate lyase was purified 43.6 folds by the following continuous treatment with ammonium sulfate fraction, DEAE-cellulose, DEAE-sephacel and Sephadex G-200 chromatography. The purified isocitrate lyase was showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified isocitrate lyase was estimated 54,000 Da by the SDS-polyacrylamide gel electrophoresis. The Km and Vmax values for isocitrate were estimated to be 0.83mM and 0.33units/ml, respectively. Activity of isocitrate lyase was inhibited by cystein-HCl and glutathione.

• Parasites, Hosts and Diseases
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• v.21 no.1
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• pp.49-57
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• 1983

The distribution and Properties of branched chain amino acid aminotransferase (EC 2.6. 1.42) was investigated in adult Fasciola hepatica. Fascicla hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of branched chain amino acid aminotransferase was measured by the method of Ichihara and Koyama (1966) . Isozyme patterns of this enzyule was also examined by DEAE-cellulose column chromatography. The results obtained were as follows; 1. The activity in homogenate was found to be 12.69 units/g wet tissue. The activity of this enzyme was relatively high compared with those in rat tissues. 2. The distribution of branched chain amino acid aminotransferase in the subcellular organelles showed that 87.8% of the activity was in cytosolic, 10.9% in mitochondrial and 1.3% was in nuclear fraction. 3. Cytosolic fraction of Fasciola hepatica contained Enzyme I, but not Enzyme II and III, of branched chain amino acid aminotransferase. Ensyme I was eluted by 50mM phosphate buffier from DEAE-cellulose column and catalyzed the transamination of all three branched chain amino acids. 4. The Enzyme I was purified about 22-folds increase in specific activity after chromatography on DEAE-cellulose. 5. The best substrate among three amino acids (leucine, isoleucine and valise) was L-isoleucine. 6. The optimal temperature of Enzyme I was $45^{\circ}C$ and the optimal pH was 8.2. 7. The Km value for leucine of Enzyme I was 4.17 mM. 8. The Km values for a-ketoglutarate and pyridoxal phosphate of Enzyme I were 0.41mM and $4.76{\times}10^{-3}{\;}mM$, respectively.

• Microbiology and Biotechnology Letters
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• v.11 no.1
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• pp.15-21
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• 1983

This experiment was carried out to optimize the condition for the enzyme production by selected strain in the basal medium, to purify the enzyme and to characterize the purified enzyme. The results obtained were as follows. 1. The optimal conditions for the $\beta$-galactosidase production were initial pH 7.0 and temperature $65^{\circ}C$. 2. Enzyme was induced by the addition of lactose and galactose, and it was intracellular enzyme. 3. The purified enzyme was obtained with the increased level of activity approximately 28.5 folds as compared with crude enzyme and the yield of 15.2% by means of DEAE-Cellulose column chromatography, Sephadex G-150 gel filtration 4. $\beta$-galactosidase from final step of purification showed a sing1e protein band on polyacrylamide gel disc electrophoresis. 5. The optimal temperature and pH of the purified enzyme were $65^{\circ}C$, pH 6.5 for the hydrolysis of lactose.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.375-380
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• 1987

D-$\alpha$-Amino-$\varepsilon$-carpolactam racemase (EC 5.1.1) and L-$\alpha$-amino-$\varepsilon$-caprolactam hydrolase (EC 3.5.2) were fractionated from cell-free extracts of Alcaligenes eutrophus A52 using ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. It was made sure that D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-$\alpha$-amino-$\varepsilon$-caprolactam by racemase, and then hydrolyzed into L-lysine by hydrolase in Alcaligenes eutrophus A52. For the conversion of D-$\alpha$-amino-$\varepsilon$-caprolactam into L-lysine by cell-free extracts of Alcaligenes eutrophus A52, the optimum temperature and pH were 6$0^{\circ}C$ and 8.5 respectively. The results showed that 0.5% D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-lysine at 55$^{\circ}C$ for 10 hr with a conversion rate of 98% by cell-free extracts containing 3.1mg of protein.

• Microbiology and Biotechnology Letters
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• v.5 no.4
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• pp.177-184
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• 1977

Arylsulfatase catalyzes the release of SO$\sub$4//sup2/- from sulfate esters of simple phenols. Arylsolfatase occurs widely in animal tissues and in microorganisms including soil bacteria. Its widespread distribution suggests that it has a rather fundamental function and environmental meaning. It has been shown previously that arylsulfatase of Klebsiella was purified and characterized. A condition of arylsulfatase synthesis was tested with several strains of Serratia. Serratia marcescens could not utilize some sugars, such as xylose, rhamnose, glucosamine and arabinose hut glucose and mannitol as a sole carbon source. However, arylsulfatase synthesis was repressed by glucose but not by mannitol. The enzyme synthesis was repressed ob inorganic sulfate and methionine, and this repression was relieved by addition of tyramine. Arylsulfatase of S. marcescen was purified by fractionation with ammonium sulfate and followed by chromatographies on DEAE-Cellulose CM-Cellulose, and DEAE-Sephadex A-25. The molecular weight of arylsulfatase was determined to be 46,000 by SDS-Gel electrophoresis and 49,000 by Sephadex G-100 column chromatography. The enzyme showed some different properties with that of K. aerogenes. The activity was maximum at pH 6.8. The Km and Vmax values for p-nitrophenyl sulfate were 2.5${\times}$10$\^$－4/ M and 20 nmoles/min/mg protein, respectively. The enzyme showed high activities toward phenyl sulfate, ο-and p-nitro phenyl sulfates, and p-nitrocatechol sulfate. The inhibition of enzyme was strongly affected by hydroxylamine, inorganic fluoride, sulfide and phosphate, but by inorganic sulfate. Like Klebsiella arylsulfatase, tyramine, octopamine, and dopamine gave signifcant inhibitory effect.

• The Journal of the Korean bone and joint tumor society
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• v.7 no.2
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• pp.41-50
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• 2001

Purpose : To investigate biochemical markers for osteosarcoma, activities of deoxyribocuclease(DNase), ribonuclease(RNase), 5'-nucleotidase, alkaline phosphatase and amylase were determined in the osteosarcoma tissue and serum of patients with osteosarcoma. Also studied were DNase, RNase in osteosarcoma tissue, isolating the enzymes from the sarcoma tissue and investigating the sarcoma specific enzymes. Materials and Methods : The experimental tissue and serum were obtained from twelve patients with osteosarcoma. The control group were obtained from the normal healthy tissue of the same patients. The tissue were centrifugalized to obtain extracts. The extracts were analized for the estimation of nucleic acid, protein contents and enzyme activities. And then each enzymes were isolated and analized by DEAE-cellulose chromatography and estimated for activities. Result : Activities of acid DNase, RNase, 5'-nucleotidase and alkaline phosphatase were significantly increased in osteosarcoma tissue. Neutral RNase in osteosarcoma tissue was shown to bo highly active, exhibiting secretory form of RNase inhibitor associated with the RNase was also increased. In the serum of patients with osteosarcoma, RNase activity was significantly increased. DEAE-cellulose column chromatographical analysis revealed that acid DNase was isolated as a single enzyme and neutral RNase as five isozymes in osteosarcoma tissue. Conclusion : The results indicated that combination of these enzymes could be used as markers for osteosarcoma. The results indicated that acid DNase and neutral RNase might play a role in genesis of sarcoma and suppression of sarcoma.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.6
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• pp.671-675
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• 2002

The present study was conducted to elucidate functional properties of sulfated polysaccharides from ascidian tunics, In physical properties of the crude polysaccharides, emulsion ability and foaminess were more excellent compared with chitin and chitosan, particular dye binding capacity was prominent. Anti-blood coagulation of partially purified sulfnted polysaccharides showed with respect to APTT (Activated partial thromboplastin time). Especially, active fraction $(F_4)$ obtained by DEAE-cellulose ion exchange chromatography showed highest activity, which was approximately $20\%$ of the activity of heparin. ACE inhibitory activity also similar to anticoagulant activity. Active fraction $(F_4)$ obtained by DEAE-cellulose ion exchange chromatography showed about $34\%$, in ACE inhibitory activity.

• Food Science and Preservation
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• v.10 no.2
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• pp.246-251
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• 2003

Pretense produced by Bacillus subtilis PANH765 was purified from culture supernatant by using ammonium sulfate fractionation DEAE-cellulose ion exchange chromatography, and gel filtration with Sephacryl S 200 HR and Sepharose CL-6B. DEAE-cellulose ion exchange column chromatography, separated the pretense into one fraction. This fraction was further purified using Sephacryl S 200 HR and Sepharose CL-6B gel titration. The molecular mass of pretense was estimated to be 35.0 kDa by the SDS-PAGE and gel filtration using Sepharose CL-6B. The results indicated that the purified pretense are monomeric proteins. Specific activity and purification folds of pretense were 657 U/mg and 4.35, respectively. The optimum temperature, optimum pit stable at a temperature range and pH ranges for the purified protease were 65$^{\circ}C$, 7.05, 50 ∼ 75$^{\circ}C$ and 6.0 ∼ 7.5, respectively. The pretense activity was decreased by the presence of PMSF and DFP, which the protease activity was increased by the presence of Na$\^$+/, K$\^$+/, Mg$\^$2＋/ and NH$_4$$\^$+/ ions.

• KSBB Journal
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• v.10 no.5
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• pp.491-498
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• 1995

In order to fraclionate ${\alpha}$-, ${\beta}$- and ${\gamma}$-cyclodextrins(CDs) from CD reaction mixture, various CD adsorbents were manufactured using fatty acids as the ligand molecules and anion exchange resins as matrix. Among several anion exchange resins, DEAE Cellulose was found to be the most suitable matrix for binding fatty acid. The binding stability between DEAE Cellulose and capric acid was tested under the various operation conditions, such as temperature, ethanol concentration, and ionic strength. Specific CD adsorbents manufactured with different chain-length fatty acids, saturated and unsaturated, were compared in terms of the recovery yield and selectivity of ${\alpha}$-, ${\beta}$- and ${\gamma}$-CDs. Stearic acid (C18, saturated) was identified as the most effective ligand for fractionation of ${\alpha}$-CD, and linoleic acid ((C18, unsaturated ) for ${\beta}$-CD. The spacer length between the matrix and ligand was required for effective adsorption of CDs, and the double bond in fatty acid molecules was also acted as an important factor determining recovery yield and selectivity. The elusion patterns of ${\alpha}$- and ${\alpha}$-, ${\beta}$-CD from column packed with stearic acid and linoleic acid CD adsorbents were also investigated at the various elusion conditions for fractionation of ${\alpha}$- and ${\beta}$-CD.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.440-446
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• 1989

Penicillium sp. CB-20 was selected for its strong polygalacturonase activity among various strains of molds found in soil. It was found that the production of polygalacturonase reached to maximum when on the wheat bran medium containing pectin as carbon source, the strain was cultured for 60 hours at 3$0^{\circ}C$. The enzyme was purified to 29.21 food by ammonium sulfate treatment, Sephadex G-25, G-15, G-150 gel filtration, DEAE-cellulose and DEAE-Sephadex A-50 ion-exchange chromatography. Yield of the enzyme purification was 2.31 %. When the purified enzyme was applied to sodium dodecyl sulfate polyacrylamide gel electrophoresis, the molecular weight was estimated 21, 000. The amino acid composition indicated relatively high contents of gultamic acid, glycine and histidine.