• Korean Journal of Food Science and Technology
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• v.38 no.6
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• pp.793-798
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• 2006

We investigated the effects on the cytotoxicity against several cancer cells of the hydrolysis and molecular weight fractionation of crude laminaran from E. bicyclis, a brown seaweed collected from Uleung island in Korea, was extracted with boiling water and then crude laminaran was prepared by ethanol precipitation of extract obtained after elimination of calcium alginate by calcium chloride. Crude laminaran was hydrolyzed by enzyme (Econase CE), acid (0.1 N HCl) and autoclaving ($121^{\circ}C$, 180 min), and the molecular weight fractions by ultrafiltration to generate molecular weight fractions. Total sugar and sulfate contents of hydrolyzed laminaran were 72.3 and 3.5% (enzyme hydrolysate), 68.5 and 3.0% (acid hydrolysate), 70.2 and 3.2% (autoclaved), and monosaccharides of which consisted of glucose (74.7-78.5%), mannose (9.9-11.5%), galactose (8.5-9.6%) and fucose (3.1-4.5%), respectively. When the cytotoxicity of hydrolyzed laminaran on SNU-1, HeLa and SW cells was evaluated by MTT assay, growth-inhibitory activity of the enzyme hydrolysate against cancer cells was higher than that of acid hydrolysate or autoclaved laminaran. Furthermore, the fraction at a molecular weight range of 10 to 50 kDa revealed higher anti-proliferative activities. The $IC_{50}$ values of 10-50 kDa fraction at a molecular weight range of 10 to 50 kDa revealed higher anti-proliferative activities. The $IC_{50}$ values of 10-50 kDa fractions on SNU-1, HeLa and SW cells were 60.4, 58.6 and 53.9 ${\mu}g/mL$ for enzymatic hydrolysate, 75.6, 73.5 and 77.4 ${\mu}g/mL$ for acid hydrolysate, and 61.7, 68.2 and 60.8 ${\mu}g/mL$ for autoclaved, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1162-1166
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• 2013

Proteins from sunflower seeds were hydrolyzed with Alcalase and Flavourzyme to isolate iron-binding peptides. The optimal hydrolysis conditions were determined. Hydrolysates were filtered under a 3 kDa membrane and iron-binding peptides separated from the hydrolysates using ion exchange and gel permeation chromatographic methods. A fraction with the highest iron-binding activity (Fe/peptide, 0.69), F22, was obtained. These results suggest that fractions isolated from sunflower seed protein hydrolysates can be applied toward the production of iron supplements.

• BMB Reports
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• v.38 no.3
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• pp.354-359
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• 2005

Open reading frame 29 (ha29) is a gene specific for Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearSNPV). Sequence analyses showed that the transcription factor Tfb2 motif, bromodomain and Half-A-TPR (HAT) repeat were present at aa 66-82, 4-76, 55-90 of the Ha29 protein respectively. The product of Ha29 was detected in HearSNPV-infected HzAM1 cells at 3 h post-infection. Western blot analysis using a polyclonal antibody produced by immunizing a rabbit with purified GST-Ha29 fusion protein indicates that Ha29 is an early gene. The size of Ha29 product in infected HzAM1 cells was about 25 kDa, which was larger than the presumed size of 20.4 kDa. Tunicamycin treatment of HearSNPV-infected HzAM1 cells suggested that the Ha29 protein is N-glycosylated. Fluorescent confocal laser scanning microscope examination, and Western blot analysis of purified budded virus (BVs), occlusion-derived virus (ODVs), cell nuclear and cytoplasmic fraction, showed that the Ha29 protein was localized in the nucleus. Our results suggested that ha29 of HearSNPV encodes a non-structurally functional protein that may be associated with virus gene transcription in Helicoverpa hosts.

• KSBB Journal
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• v.10 no.5
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• pp.589-597
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• 1995

Protein-bound polysaccahrides(PBP) were isolated, purified, and characterized from the sawdust media after harvesting the fruit body of Flammulina velutipes. The yield of the crude PBP(Fr.CA) extracted from the sawdust media, was 0.367% relative to the original sawdust media. The total sugar and protein contents of Fr.CA were 19.8% and 23.8% respectively. Using the membrane filtration, the fraction of which the molecular weight is over 300 kDa(Fr.A) was isolated from the Fr.CA and the yield was 44.6% relative to the Fr.CA. This result indicates that high molecular PBP is the dominant components of the Fr.CA. The Fr.A was separated into three fractions (Fr.A-1, Fr.A-2 and Fr.A-3) whose yields are 5.8%, 8.5% and 13.2% respectively. These fractions were further purified using gel filtration, obtaining a single peak in each fraction that considered as pure PBP Among them, the yield of Fr.A-1-${\alpha}$ was 20.9% relative to the Fr.A-1, and the molecular weight was 800 kDa. Monosaccharide components such as glucose, galactose, mannose, xylose and fucose could be detected all fractions by a HPLC analysis. Especially in the Fr.A-1-${\alpha}$ fraction, the content of glucose and galactose appeared to be high.

• BMB Reports
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• v.39 no.5
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• pp.546-555
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• 2006

An acidic polysaccharide fraction that had high anti-complementary activity was isolated from the stems of Grey Mangrove in 0.15% yield. The final fractions was designated HAM-3-IIb-II. The polysaccharide fraction appeared to be homogenous by high performance size exclusion chromatography with an estimated molecular weight of 105 kDa. The isolated polysaccharide is more effective than polysaccharide K (PSK) in its anti-complementary activity at 58 ${\mu}g/ml$ of PSK and 23 ${\mu}g/ml$ of HAM-3-IIb-II that inhibit 50% of complement activity in the complement fixation assay. Structural studies indicated that HAM-3-IIb-II was rich in galacturonic acid along with arabinose, galactose and rhamnose, characterizing a pectin-type polysaccharide, which was also confirmed by FT-IR spectrum. The presence of rich neutral sugar side chains of arabinogalactans may have contributed to the expression of high activity. Traditionally, this mangrove plant is used for medicinal purposes and it appears to have some scientific applications.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.10
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• pp.1046-1054
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• 2006

The raw water DOC contained 39.3% of hydrophilics, 42.9% of hydriophobic, and 17.8% of transphilic. The hydrophobic fraction in this raw water was mostly fulvic acid. Fulvic acid comprised of 62% and the rest was humic acid(38%). There was more carboxylic acid functional group(64%) than phenolic group(36%). HPI-N and HPI-C comprised of 17% and 22% in the hydrophilic portion, respectively. The fouling mechanisms on the membrane surface and into its porous structure were analyzed in terms of several kinetic models. In order to analyze the fouling kinetics, the various kinetic models described in this paper were used to fit the experimental results. The kinetic models and kinetic constants obtained for each operation condition. The permeate flux was rapidly declined by simultaneous pore blocking and cake formation. Also, the permeate flux declined with decreasing internal pore size resulted from organic deposition into the membrane pore. The results of the membrane fouling test using UF membrane according to NOM fractions. HPI-N caused more fouling than HPI-C. Humic acid caused more fouling than fulvic acid probably due to higher adsorption capacity. Since humic acid has higher adsorption capacity than fulvic acid, it would be more adsorbed onto the membrane pores.

• Food Science and Preservation
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• v.24 no.4
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• pp.542-549
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• 2017

In this study, a peptide exhibiting antioxidant activity was isolated from sandfish (Arctoscopus japonicus) roe hydrolysate (SRH) in order to evaluate their practical uses as materials for manufacturing functional foods. The A. japonicus roe protein was hydrolyzed using Collupulin MG, and isolation of antioxidant peptide was performed using ultrafiltration (UF), prep-HPLC, and RP-HPLC. The SRH with a molecular weight below 3 kDa constituted about 38% of the whole hydrolysate, and the fraction with a molecular weight below 3 kDa showed significantly greater antioxidant activity compared to the original SRH and other fractions. The isolation fold of the antioxidant peptide isolated from SRH throughout the four-step procedure was 7.11-fold, and protein yield was 14.8%. The DPPH radical scavenging activity of isolated antioxidant peptide was above 90% at a concentration of 1.0 mg/mL, which was similar to that of the Trolox at a concentration of 0.1 mg/mL. These results suggested that the antioxidant peptide derived from A. japonicus roe could be a useful additive for producing functional foods and protein supplements. However, it is necessary to perform further study the structural characteristics of this antioxidant peptide isolated from A. japonicus roe.

• BMB Reports
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• v.37 no.3
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• pp.275-281
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• 2004

Rhodanese was isolated and purified from the cytosolic fraction of liver tissue homogenate of the fruit bat, Eidolon helvum, by using ammonium sulphate precipitation and CM-Sephadex C-50 ion exchange chromatography. The specific activity was increased 130-fold with a 53% recovery. The $K_m$ values for KCN and $Na_2S_2O_3$ as substrates were $13.5{\pm}2.2\;mM$ and $19.5{\pm}0.7\;mM$, respectively. The apparent molecular weight was estimated by gel filtration on a Sephadex G-100 column to be 36,000 Da. The optimal activity was found at a high pH (pH 9.0) and the temperature optimum was $35^{\circ}C$. An Arrhenius plot of the heat stability data consisted of two linear segments with a break occurring at $35^{\circ}C$. The apparent activation energy values from these slopes were 11.5 kcal/mol and 76.6 kcal/mol. Inhibition studies on the enzyme with a number of cations showed that $Mg^{2+}$, $Mn^{2+}$, $Ca^{2+}$, and $Co^{2+}$ did not affect the activity of the enzyme, but $Hg^{2+}$ and $Ba^{2+}$ inhibited the enzyme.

• The Korean Journal of Mycology
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• v.38 no.1
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• pp.57-61
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• 2010

For development of anti-obesity nutraceuticals from mushrooms, new anti-obesity lipase inhibitor from Phellinus linteus was purified by systematic solvent extraction, TLC and HPLC and characterized. Methanol extract from P. linteus most effectively inhibited(72.5%) porcine pancreatic lipase and ethyl acetate fraction showed the highest inhibitory activity in the systematic solvent extraction. A lipase inhibitor from the ethyl acetate fraction was purified through following steps including 3 times silica gel chromatography and preparative HPLC. The purified lipase inhibitor was a yellowish geen powder and its $IC_{50}$ value was 175 ng. Its molecular weight by MALDI-TOF-MS was 523.06 Da and showed maximal absorption spectrum at 225.1 nm.

• Korean Journal of Microbiology
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• v.39 no.4
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• pp.223-229
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• 2003

The purpose of this work was to perform the characterization of NAD(P)H-nitroreductase isolated from Stenotrophomonas sp. OK-5 capable of degrading 2,4,6-trinitrotoluene (TNT). Initially, NADP(H)-nitroreductase by a series of purification processes including ammonium sulfate precipitation, DEAE-sepharose, andQ-sepharose was prepared. From samples harvested from fraction collector, three different fractions (I, II & III)having the enzyme activity of NAD(P)H-itroreductase were detected. Specific activities of three fractions I, II,and III of NAD(P)H-nitroreductase were determined to approximately 5.06 unit/mg, 4.95 unit/mg and 4.86 unit/mg, and concentrated to 10.5, 9.8, and 8.9-fold compared to crude extract, respectively. Among these three fractions,the fraction I of NAD(P)H-nitroreductase demonstrated the highest specific activity in this experiment. Several factors affecting on the enzyme activity of NAD(P)H-nitroreductase (fractions I, II & III) were investigated.The optimum temperature of all NAD(P)H-nitroreductase (fractions I, II & III) was 30oC, and the optimal pH was approximately 7.5. Metal ions such as Ag+, Cu2+, Hg2+ inhibited approximately 80% enzyme activity of all NAD(P)H-nitroreductase, and the enzyme activities were decreased about 30-40% inhibition in the presence of Mn2+ or Ca2+. However, Fe3+ showed stimulatory effect on the enzyme activity. The molecular weights of NAD(P)H-nitroreductase (fractions I, II & III) were measured about 27 kDa on the SDS-PAGE.