• Applied Biological Chemistry
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• v.30 no.2
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• pp.169-178
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• 1987

The extracellular and intracellular inulases from Kluyveromyces marxianus were purified and characterized. The maximum production of both inulases was achieved at stationary phase in a pH-controlled medium at pH 5 with yeast nitrogen base as organic nitrogen source. Each enzyme was concentrated by tannic acid precipitation and separated into two fractions by DEAF-cellulose chromatography. Electrophoretic analysis showed that the four fractions had three glycoprotein bards each. Only main glycoprotein band, however, had both inulase and invertase activities. There were no significant differences between two enzymes in the optimum pH and temperature. But the intracellular inulases had higher heat stability and less affinity toward inulin than the extracellular enzymes do. All the purified enzymes were considered to be exo-inulases using hydrolyzate analysis with TLC.

• Applied Biological Chemistry
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• v.15 no.1
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• pp.27-33
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• 1972

The alkaline protease was isolated from the culture material of monascus sp. on wheat bran culture. The crude purification of this enzyme was extracted with distilled water and precipitated with ammonium sulfate of 0.5 saturation. And, the activity of this enzyme was determind very strongly by folin's colorimetric method. The optimal pH of this enzyme was ranging from pH 10 to 13 and the optimal temperature was $50^{\circ}C$. The pH stability was ranging from pH 5 to 12 and the enzyme activity was not inactivated by heat treatment in lower temperature than $40^{\circ}C$. The enzyme was protected from heat denature by the treatment of $Pb^#$, $Ba^#$, $Co^#$, $Zn^#$, and $Cu^#$, but was inactivated with $Hg^#$, $Fe^#$ strongly. Moreover, one of these metal ions, the cupper ion, has a strong protective activity on enzyme heat denature. And, it was not effected by treatment of EDTA.

• KSBB Journal
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• v.10 no.1
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• pp.9-14
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• 1995

Continuous production of fructooligosaccharides from sucrose by a dual immobilized enzyme system of fructosyltransferase and glucose isomerase was studied in a column reactor. The optimal temperature and pH of the immobilized fructosyltransferase were $65^{\circ}C$ and 5.5, respectively. The activity of glucose isomerase was favorable as temperature and pH were increased within the ranges examined. However, both the immobilized enzymes were thermally unstable over $5^{\circ}C$, suggesting that long-term operation of the dual immobilized enzyme column should be carried out below $50^{\circ}C$. The optimum packing ratio of fructosyltransferase to glucose isomerase was found to be around 5/3. Under the optimized reaction conditions, the dual enzyme column was successfully operated for 40 days without any loss of initial enzyme activities, yielding 66% of fructooligosaccharides. Furthermore, the relative sweetness of fructooligosaccharides produced by a dual emzyme system was enhanced by 6% compared with that of fructosyltransferase alone.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.257-263
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• 1998

Alcaligenes eutrophus was successfully recovered from high cell density broth by pre-treatment with Fe-based coagulants. An inorganic coagulant, Fe$_2$(SO$_4$)$_3$, and a polymerized coagulant, Ferix-3, were used. Good coagulation was observed in broad pH range of 3 to 13, the floe size was increased with increasing pH of culture broth. The optimum pH of fermentation broth for cell recovery was 10 to 13. The optimum coagulant dosages to recover cells with 95% cell recovery were increased with increasing cell concentration. Optimal coagulant dosage was lower when the polymerized coagulant was used rather than the inorganic coagulant. The coexistence of NH$_4$$\^$+/ was increased coagulant requirement, and the coagulant requirement was 0.066g Fe$_3$$\^$+//g NH$_4$$\^$+/.

• Parasites, Hosts and Diseases
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• v.30 no.3
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• pp.191-200
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• 1992

A proteolytic enzyme was purified from the tissue extract of spargana (plerocercoids of Spirometra erinacei) by DEAE-Trisacryl M ion exchange chromatography and thiopropyl-sepharose affinity chromatography resulted in a 21-fold purification. The proteinase activity was assayed with a synthetic fluorescent substrate, carbobensoxy-phenylalanyl-7-amiso-4-trifluoromethyl-coumarin. SDS-polyacplamide gel electrophoresis of the purified materials revealed a single 28,000 dalton band. Inhibitor profiles of the band indicated that it belonged to cysteine endopeptidases. It exhibited identical pH curves with optimum at pH 5,5, and 50% activity from pH 4.7 to 8. It could completely degrade collagen chains to three identical products. It also showed some activity on hemoglobin. Furthermore, the band on immunoblots was reactive to the sera of sparganosis patients. These results suggest that the proteolytic enzyme belongs to cysteine proteinase which plays a role in the tissue penetration. Also it may be used as the antigen for diagnosis of active sparganosis.

• Korean Journal of Food Science and Technology
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• v.30 no.2
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• pp.293-298
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• 1998

Allithiamine was synthesized when thiamine was mixed with fresh garlic extract made with ethanol at a alkali medium. Allithiamine was isolated and purified using solvents such as ethyl acetate, diethyl ether and benzene. Purified allithiamine was identified by determination of melting point, elemental analyzer and LC/Thermospray/Mass-spectrometer. On the other hand, synthetic conditions of allithiamine from fresh garlic, pH, temperature and ratio of garlic to ethanol were investigated. Synthetic rates of allithiamine under alkali conditions were rapidly increased while those under acidic conditions very slowly increased. The synthetic rates of allithiamine increased as temperature increased, but decreased above $70^{\circ}C$ as reaction time increased. There was no significant difference in synthetic rate of allithiamine when garlic was mixed above 4 times of ethanol. Therefore, optimum condition of pH, temperature and ratio of garlic to ethanol for synthesis of allithiamine were 8, $60^{\circ}C$ and 1 : 4, respectively.

• Microbiology and Biotechnology Letters
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• v.7 no.3
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• pp.141-147
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• 1979

Naringinase from Atpergillus nidulans was immobillized on DEAE-Sephadex A-25 and its characteristics were studied. The optimal conditions for the preparation of the immobilized enzyme were as follow; optimal pH, incubation time and the suitable amount of enzyme were 6.0, 30 min. and 110 units per gram of the dried ion exchage resin, respectively. The optimal pH of the immobilized enzyme was higher than that of the native enzyme. The optimal temperature increased from 4$0^{\circ}C$ to 5$0^{\circ}C$. The heat and pH stability of the immobillized enzyme were better than those of the native enzyme. No significant difference in the Michaelis constant was detected. Activation energy of the immobilized enzyme was 7.96 Kcal/mole, and the apparent Michaelis rate equation was used to describe the action of this material. The degree of hydrolysis was dependant on the flow rate at low rate of perfusion through the column. As the flow rate increased, the value of the apparent Km decreased.

• 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.

• Applied Biological Chemistry
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• v.27 no.4
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• pp.271-277
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• 1984

Molecular weight of Exo-maltotetraohydrolase produced by Pseudomonas stutzeri IAM 12097 was estimated to be approrimately 63,000 and 60,000 with SDS-polyacrylamide gel electrophoresis and Sephadex-G-100 gel filtration, respectively. The isoelectric point was appeared to be pH 4.8. Optimum pH, the stable pH range and optimum temperature of this enzyme were pH 6.6, $pH6.0{\sim}10.5\;and\;45{\sim}50^{\circ}C$. The enzyme was stable below $40^{\circ}C$ and was rapidly inactivated above $55^{\circ}C$. This enzyme was inactivated completely by $Ag^+,\; Hg^{++},\;I_2$ and ${\beta}-cycoldextrin$, and slightly by EDTA, ${\rho}-CMB$ and IAA. Michaelis constant(Km) of this enzyme toward soluble starch, amylose and amylopectin were 7.70mg/ml, 6.17mg/ml, 5.56mg/ml, respectively.

• KSBB Journal
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• v.14 no.6
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• pp.661-664
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• 1999

Proteins were extracted from an aqueous phase with reversed micelles. The effect of pH, and salt concentration on the solubilization of lysozyme in AOT/isooctane solution was studied to explore the potential for employing this solvent system in the large-scale recovery and concentration of proteins using liquid extraction. For pH values below the isoelectric point, pl of the protein, solubilization was high, probably owing to strong electrostatic interactions between the positively charged proteins and the anionic surfactant heads forming the inner micelle wall. At low ionic strength complete solubilization of the protein was observed. A pH higher than the pl of lysozyme and a salt concentration lower than that of the water pool were required for the recovery aqueous phase to ensure the back extraction of lysozyme from the AOT reversed micelles.