• KSBB Journal
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• v.8 no.3
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• pp.287-294
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• 1993

For the purpose of combining the purification processes for several biologically active proteins form human urine, an efficient integrated fractionation procedure has been investigated. The procedure was started by concentration with ultrafiltration and pH precipitation followed by a selectable combination of chromatography on gel filtration, adsorption, ion exchanger, affinity, and reverse phase column. By this process, the purified urokinase, epidermal growth factor and albumin migrated as a single band on SDS-polyacrylamide gel electrophoresis and were fully active. The recoveries of these purified proteins were 48%, 17%, and 46%, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.2
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• pp.175-179
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• 1997

Changes of protein contents and chromatogram patterns by gel filtration chromatography was investigated for the purpose of studying changes of Proteins during softening of persimmon and jujube fruits. Contents of water-soluble and salt-soluble proteins were increased during softening of persimmon and jujube fruits, but that of cell wall-bound proteins was decreased. After performing a gel filtration of water-soluble protein, one peak was separated in mature persimmon fruits and three peaks in soft persimmon fruits. In the case of jujube fruits, there were three peaks in both of mature and soft fruits. Pattern of salt-soluble and cell wall-bound proteins by gel filtration chromatography hardly changed during softening. During softening of two fruits, the contents of water- soluble and salt-soluble proteins appeared to be increased on the same fractions with the decreasing in content of cell wall-bound proteins.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.132-138
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• 1994

L-Ascorbic acid-producing enzyme in Neurospora crassa was found to exist in mitochondria and the activity of this enzyme was increased by the addition of D-fluconno-${\gamma}$-lactone or L-gulono-${\gamma}$-lactone in the media. L-Ascorbic acid-producin enzyme in N. crassa has been purified with ammonium sulfate precipitation. DEAE Sepharose CL-6B ion exchange chromatography. Sephacryl S-200 gel filtration chromatography and Reactive yellow 3-agarose dye affinity column chromatography. The specific activity of this enzyme was increased to 239.6 fold and the yield was 2.1%. The molecular weight of the native enzyme was 150.000 dalton when it was estimated with Sephacryl S-200 gel filtration chromatography. Its molecular weight appeared as 75.000 dalton by SDS-polyacrylamide gel electrophoresis. which suggested that this enzyme was consisted with two identical subunits. The optimal pH for this enzyme was 9.0 and the $K_m$ value for D-galactono-${\gamma}$-lactone was 0.073 mM.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.313-320
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• 1989

Rhodotorula glutinis K-24 was found to produce internal, cell wall bound, and external invertase. Internal invertase was purified by column chromatographies on DEAE-Sephadex A-50, Sp-sephadex C-50, gel filtration on Sephadex G-200 and isoelectric focusing. Cell wall bound invertase was partially purified by the following procedures; column chromatography on DEAE-Sephadex A-50 and gel filtration on Sephadex G-100. Optimum pH and temperature for enzymatic activities of internal and cell wall bound invertase were pH 3.0 and 6$0^{\circ}C$, respectively. Both enzymes were inhibited by HgC1$_2$, AgNO$_3$, MnSO$_4$, and sodium dodecylsulfate. The molecular weights of internal and cell wall bound invertases were estimated to be 310,000 and 61,000, respectively. Other physicochemical properties of the both enzymes were similar.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.2
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• pp.180-185
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• 1997

This work was carried out to investigate change of polygalacturonase activities and polygalacturonase patterns by gel filtration chromatography during softening of persimmon and jujube fruits. During softening of two kinds of fruit, polygalacturonase activities of water-soluble and salt-soluble proteins were increased, but that of cell wall-bound proteins was decreased. In water-soluble and salt-soluble proteins of persimmon fruits, two peaks of polygalacturonase activity were separated in mature stages, but one peak in soft stages. During softening of those fruits, the peaks of polygalacturonase activity in water-soluble and salt-soluble proteins appeared on the same fraction with the peaks of polygalacturonase activity in cell wall-bound proteins.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.4
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• pp.351-358
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• 2013

A protease inhibitor from fish eggs was fractionated using chromatographic methods. The fractionation efficiency was evaluated in terms of specific inhibitory activity (SIA, U/mg), purity (fold), total inhibitory activity (TIA, U), and recovery (%). The protease inhibitor (PI) from egg extracts of skipjack tuna (ST Katsuwonus pelamis), yellowfin tuna (YT Thunnus albacares) and Alaska pollock (AP Theragra chalcogramma) was fractionated using Sephadex G-50 gel filtration and DEAE-Sepharose CL-6B anion exchange chromatography based on protein size exclusion and net charge, respectively. Fractions exhibiting strong inhibitory activity were contained in the 30-50 kDa fraction on gel filtration and in the range of 0.4-0.7 M NaCl gradient fraction on anion exchange chromatography. The respective TIA and percent recovery of the fraction obtained with gel filtration toward trypsin and $N{\alpha}$-benzoyl-L-arginine-p-nitroanilide (BAPNA) were 2,758.7 U and 29.6% for ST, 1,005.5 U and 25.6% for YT, and 1,267.5 U and 26.0% for AP. Gel filtration chromatography was more effective at fractionating PI than using ion exchange chromatography. These results suggest that fish eggs act as serine protease inhibitors and might be useful for protease inhibition in foodstuffs.

• 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.

• Applied Biological Chemistry
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• v.21 no.3
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• pp.137-143
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• 1978

Xanthine oxidase from bovine thyroid glands was found to contain FAD, molybdenum and iron in a ratio 1:0. 36:1. 6. The molecular weight of the thyroid enzyme was similar to that of the milk enzyme when estimated by gel filtration and polyacrylamide gel electrophoresis. The optimum pH for the enzyme activity was 7.8. The pH of the isoelectric point was determined to be 6.2 by electrofocusing. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis experiment indicated that the enzyme was dissociated into subunits and that the molecular weight for the smallest subunit was 65,000 daltons. Absorption spectra were dissimilar between milk and thyroid xanthine oxidase.

• Korean Journal of Animal Reproduction
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• v.19 no.4
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• pp.251-258
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• 1996

These studies were carried out to investigate the effect of gonadotropins (GTH), fetal calf serum (FCS), porcine follicular fluid (pFF) and FCS and pFF fractions obtained by the gel filtration on in vitro maturation of porcine follicular fluid. When the oocytes were cultured in TCM-199, the maturation rate was higher in pFF than in FCS in both with or without GTH and in pFF the maturation rate was higher in with GTH than in without GTH. In case of without GTH, pFF increased maturation rates in TCM-199, but not in Whitten's medium (WM). When the oocytes were cultured in WM supplemented with FCS fractions, the maturation rate(51.6%) of oocytes was significantly (P<0.05) higher in fraction B (about 30∼70 kDa) than in control, FCS and other fractions. When oocytes were cultured in WM supplemented with pFF fractions, fractions B (about 30∼70 kDa) and D (about 1∼10 kDa) were significantly (P<0.05) higher than in control, pFF and other fractions. In conclusiion, the addition of gonadotropins into the maturation media was effective for oocyte maturation. The addition of pFF was more effective than addition of FCS for maturation of porcine oocytes in vitro. And fraction B from FCS and fractions B and D from pFF was effective for oocyte maturation.

• BMB Reports
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• v.28 no.3
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• pp.197-203
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• 1995

Famesyl protein transferase involved in the first step of post-translational modification of $p21^{ras}$ proteins transfers the famesyl moiety from famesyl pyrophosphate to a cysteine residue in $p21^{ras}$ proteins. The enzyme was first purified 30,000-fold from bovine testis by use of 30~50% ammonium sulfate fractionation, DEAE-Sephacel ion exchange chromatography, Sephacryl S-300 gel filtration chromatography, Sephacryl S-200 gel filtration chromatography, and hexapeptide (Lys-Lys-Cys-Val-Ile-Met) affinity chromatography. The molecular weight of the purified enzyme was estimated to be ~100 kDa by gel filtration and SDS-polyacrylamide gels showed two closely spaced bands of ~50 kDa protein. These indicate that the enzyme consists of two nonidentical subunits, a and 13, which have slightly different molecular weights. The enzyme was inhibited by hexapeptide (Lys-Lys-Cys-Val-Ile-Met), which acted as an alternative substrate that competed for famesylation. Kinetic analysis by measuring initial velocities showed that famesyl protein transferase is a very slow enzyme. EDTA-treated famesyl protein transferase showed little activity with $Mg^{2+}$ or $Zn^{2+}$ alone, but required both $Mg^{2+}$ and $Zn^{2+}$ for the catalytic activity.