• The Korean Journal of Mycology
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• v.27 no.2 s.89
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• pp.100-106
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• 1999

The polysaccharides, intracellular and extracellular, extracted from the liquid culture of the Agrocybe cylindracea were purified and characterized. The mycellial cellular productivity of Agrocybe cylindracea was proved to be almost 2 folds in the shaking culture compared to the standing culture. These polysaccharides were purified by the DEAE-cellulose ion exchange chromatography and the Sepharose 2B size exclusive gel filteration. The two purified fractions of extracellular polysaccharides, ACEPDG and ACEPAG, contained 75.8% and 65.4% total sugar respectively. The total sugar content of ACIPDG and ACIPAG, the two purified fractions of intracellular polysaccharides, were 89.2% and 54.2% respectively. The molecular weights range of all the substances were estimated to be above 100,000, from 300KDa of ACEPDG to 600 KDa of ACIPAG. The results of sugar analysis by HPLC showed that the sugar part of ACEPDG was consisted of glucose and inositol. The ACIPDG, ACEPAG and ACIPAG contained three kinds of monosaccharides, glucose, fructose and inositol.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.4
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• pp.687-694
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• 1996

Myrosinase was purified from Korean mustard seed(Brassica juncea) by a sequential process of DEAE-cellulose, concanavalin A-sepharose, and Superose 6 chromatography. The molecular weight of puri-fied myrosinase(II-2) determined by SDS-polyacrylamide electrophoresis was 67KD. About a 248-fold purification for myrosinase II-2 was obtained after Superose 6 chromatography. Optimum pH of the myrosinase was 7.0 and optimum temperature of the enzyme was $3^{\circ}C.$ The enzyme was stable at pH 7.0, and below $30^{\circ}C.$ Cu, Hg and Fe ion significantly inhibited the enzyme activity, but ascorbic acid enhanced, resulting in a maximum activity by 1mM ascorbic acid. Among tile ascorbic acid ana-logues, dehydroascorbic acid inhibited the enzyme activity, whereas others showed a little effect. Reducing agents such as 2-mercaptoethanol and dithiothreitol inhibited the enzyme activity, but the reducing agents with ascorbic acid was enhanced enzyme activity.

• Korean Journal of Soil Science and Fertilizer
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• v.7 no.4
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• pp.209-213
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• 1974

The isozymes of glutamic oxalacetic transaminase from young rice root were isolated by DEAE-cellulose column chromatography, and investigated some of their enzymic properties. Obtained results were summarized as follows: 1. 93 percent of GOT activity in rice root existed in supernatant fraction, and their specific activity exibited the same pattern. 2. The rice root contained two types of GOT isozyme; cationic and anionic GOT. It seemed that both of them were distributed in supernatant fraction. 3. GOT isozymes of rice root on the pH dependance showed their maximum activity at 7.4. 4. On the stability of GOT isozymes for temperature, anionic GOT was more unstable than cationic GOT. 5. Apparent Michaelis constant for L-aspartate of GOT isozymes from rice root showed 12-14mM in the cationic GOT and 16mM in the anionic GOT, respectively.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.224-229
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• 2003

A keratinolytic protease was purified from the culture medium of Pseudomonas sp. KP-364 by use of an assay of the hydrolysis of feather keratin. Membrane ultrafiltration and DEAE-cellulose ion-exchange resin and Sephadex G-150 gel chromatographies were used to purify the enzyme. The specific activity of the purified keratinolytic protease relative to that in the original medium was approximately 72-fold high. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Sephadex G-150 chromatography indicated that the purified keratinolytic protease is monomeric and has a molecular weight of 36 kDa. The optimal pH and temperature of the keratinolytic protease activity were 6.6 and 37 C, respectively, and the keratinolytic protease was relatively stable at pH value from 3.0 to 10.0 at 37 C for 1hour. The keratinolytic protease was inhibited by EDTA and EGTA, indicating that the keratinolytic protease was a kind of metalloprotease that require Li+ for cofactor.

• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.285-290
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• 2008

Rahnella aquatilis strain AY2000 produces an anti-yeast substance (AYS), however activity of the AYS has a declining tendency during storage. To investigate what has been decreased activity of the AYS, the AYS was treated with various physicochemical agents in this paper. The activity of AYS was decreased by heat treatment. Thiol reagent such as $\beta$-mercaptoethanol or dithiothreitol was also another factor decreasing the activity of AYS. However, pH, EDTA, and NaCl were not factors decreasing the activity of AYS. Use of methanol to precipitate the AYS was also decreased the activity of AYS. The activity of AYS was not lost after Sepharose S-400 gel filtration. However, the AYS activity was completely lost, when a polysaccharide and a unknown substance (230 nm absorption) among components of the AYS was separated by DEAE-cellulose chromatography. MIC of the AYS against S. cerevisiae was usually determined at $7.8-15.6{\mu}g/ml$.

• Applied Biological Chemistry
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• v.33 no.1
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• pp.79-86
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• 1990

Cyclodextrin hydrolase from Bacillus stearothermophilus KFCC 21203 was purified and the properties of the purified enzyme were investigated. The enzyme was purified 15 folds with 77 % recovery by ammonium sulfate fractionation, DEAE-cellulose chromatography, and Ultro AcA 34 gel filtration. The specific activity and the molecular weight of the enzyme were 1.30 units/mg protein and about 29,500, respectively, The maximum activity of the enzyme was shown at $55^{\circ}C$ and pH 5.5. However, stable temperature and pH were $40^{\circ}C$ and $5.0{\sim}8.0$, respectively. The Km value for ${\gamma}-cyclodextrin$ was $3.78{\times}10^{-3}$ M. The degradation activity of the enzyme was selectively high for ${\gamma}-cyclodextrin$, and very low for ${\beta}-cyclodextrin$, but not for ${\alpha}-cyclodextrin$. The decomposed products of ${\gamma}-cyclodextrin$ were mainly glucose and maltose, and a little mlatotriose. The activity of the enzyme was very high for amylose, potato starch, corn starch, amylopectin and maltooligomer, and relatively high for glycogen and dextrin. The decomposed products of them were mainly glucose and maltose.

• Applied Biological Chemistry
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• v.13 no.3
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• pp.231-235
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• 1970

1. Purification of amylase system produced from Humicola sp. by a submerged culture eras carried out. 2. By DEAE-Cellulose column chromatography amylase system was separated into two fractions eluted at 0.05M and 0.5M phosphate buffer solution of pH 6.0. 3. The saccharogenic amylase was mostly composed of. the fraction of 0.05M phosphate buffer solution of pH 6.0 while the dextrinogenic amylase was perseted in fraction of 0.5M phosphate buffer solution of pH 6.0 4. It was found that the optimum pH of this saccharogenic amylase was within the range of from 4.5 to 5.5, stable pH was within the range of from 4.0 to 9.0 and optimum temperature was $60-65^{\circ}C$. This amylase was stable at $70^{\circ}C$ for ten minutes but completely inactivated $80^{\circ}C$ above.

• Microbiology and Biotechnology Letters
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• v.19 no.4
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• pp.405-412
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• 1991

The enzymes were immobilized by treating the microbial cells in 0.05% chitosan and 0.28% glutaraldehyde solution. The activity of immobilized cell was about 535 IGIC/g. Glucose isomerase was purified by 6.5 times after homogenization using 60% $(NH_4)_2S0_4$ fractionation, DEAE-cellulose and Sephadex G-150 gel filtration. The molecular weight of enzyme was about 140,000 when it was measured by HPLC and the purified enzyme had only one band by electrophoresis. It showed good enzyme activity at pH 7.5 and $75^{\circ}C$. The optimum conditions for enzyme reactions were shifted to pH 7.0 and $80^{\circ}C$ when the enzyme was immobilized. The enzyme reaction was activated by the addition of 5~10 mM magnesium ion and the thermostability was improved by the addition of 0.25 mM cobalt ion. The enzyme activity was competitively inhibited by sugar alcohols.

• Journal of Sericultural and Entomological Science
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• v.30 no.2
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• pp.96-104
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• 1988

Vitellin, the major yolk protein of the silkworm, Bombyx mori was pruified, and its immunological properties and the quantitative changes during embryogenesis were studied. The ovary transplantation into male hosts was also carried out to find its effect on the yolk protein synthesis. The pupal vitellogenin and the egg vitellin of Bombyx mori were purified by DEAE-cellulose column chromatography. These two female specific proteins showed the same mobility in the polyacrylamide gel electrophoresis and the same reaction in the double immunodiffusion test. The immunological identity was also observed between the vitellins of Bombyx mori and Bombyx mandarina. The rudimentary ovaries transplanted into the male hosts of silkworms produced eggs without vitellin, indicating that the yolk precursors synthesized in other female organ beyond the ovary were necessary to produce vitellins. The major yolk protein, vitellin was disintegrated and utilized mostly during late stage of embryogenesis. It was different characteristics from the egg specific protein, which was utilized continuously from the early embryonic stage.

• Korean journal of applied entomology
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• v.34 no.2
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• pp.140-146
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• 1995

Changes in protein content during oocyte development was measured and egg-specific protein was characterized from the eggs in Lucilia illustris. During normal development ovarian protein was rapidly increased at 72hr and reached maximum at 96hr after a protein meal, when the eggs were fully matured. Purified protein from the ovaries by gel filtration of DEAE-cellulose an Sephacryl S-200 was loaded on 7.5% native polyacrylamide gel electrophoresis and identified at ${R}_{f}$ 0.4 as egg-specific protein, which has a mol. wt of 110,000. A total of 13 amino acids in th egg-specific protein was identified and expecially asparagine, glutamic acid, and tyrosine were highly concentrated. Five fatty acids were also identified. It is suggested that there is a specific protein in the eggs of L. illustris except yolk protein synthesized and secreted by fat body.