• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.2
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• pp.240-248
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• 1996

To clarify the potentiality of sea cucumbers as dietary food, the effects of those glycoprotein on dietary proteins and physicochemical properties of those proteins were studied. Crude glycoprotein was efficiently extracted using 20mM sodium phosphate beffer(pH 7.0) and by salting out with 80% ammoniym sulfate saturation. The fractions obtained through the DEAE-cellulose ion exchange chromatography was identified as glycoprotein by Schiff's reagent and SDS polyacrylanide gel electro-phoresis. The yields of each glycoprotein from the three kinds of sea cucumbers were 0.814(red), 0.184(blue) and 0.232(black) and the molecular weights of the glycoproteins subunits were ranged from 20,000 dalton(blue and black) to 29,000 dalton(red), respectively. The electrophoretic patterns of the glycoprotein isolates were similar to each other and any significant difference in amino acid pattern was observed. Predominant arnino scids were Asx(aspartic acid and asparagine) and Glx(glutamic acid and glutamine) ; in contrast, histidine and methionine were below 2% as compared to total amino acids. water holding capacities of the glycoprotein isolates from red, blue and black cucumbers were equally 100% and emulsion activities ranged from 53% to 64%. In addition the emulsion stabilities were 7.04, 1.37 and 2.44, respectively. In vitro digestibility of some proteins(casein, SPI and squid) was decreased as increasing the level of the freeze dried sea cucumber powder and glycoprotein isolates. But squid protein was not affected.

• Parasites, Hosts and Diseases
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• v.28 no.2
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• pp.91-100
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• 1990

Paragonimus westermani is a tissue migrating parasite in the early stage until arriving at lung, and most of the parasites spend their life spans there. Considerable immune responses including activation of macrophages are taken place during the residence of parasites in the host. However, concerning the immunologic defense mechanisms of the host against this parasite, only a few document is available so far. In this study, the cytotoxic effect of peritoneal macrophages under the presence of antibody and/or complement against metacercariae of F. westermani was investigated in vitro. Metacercarlae were collected from the crayfish, Cambaroides similis and hatched out in Tyrode solution (pH 7.4). Plastic adherent cells from normal or infected rat (Wistar) peritoneal exudates were used as experimental macrophages. Polyclonal antibodies were obtained from infected rats and a cat. Cat IgG was fractioned with ion exchange chromatography. Fresh rabbit complement was used according to experimental scheme. Various combinations of peritoneal macrophages, normal or infected rat serum, complement and cat IgG were incubated at $36^{\circ}C$ in 5% $CO_2$ incubator for 6, 14, 24 and 48 hours. The results obtained were as follows: 1. P. westermani infection activated peritoneal macrophages non-specifically and this activation induced increases of cell adherence and cytotoxicity on metacercariae. 2. In the presence of infected rat serum the antibody.dependent cell-mediated cytotoxicity of peritoneal macrophages on metacercariae was significantly increased and showed a peak at 6-hour incubation. But the cytotoxic effect was markedly reduced after inactivation of complement and heat.labile IgE antibody by the heating of infected serum at 56$^{\circ}C$ for 30 minutes. 3. The highest cytotoxic effect (100%) of concomitant incubation with IgG and complement showed 24 hours after incubation, although cell adherence was relatively low at 6-hour incubation and 0% at 24-hour incubation. 4. Coordinative functions of complement with serum and IgG were effective in cell adherence and in cytotoxicity, but it is not clear the independent role of complement on the macrophage- mediated cytotoxicity in this study- With these results it is assumed that P. westermani infection can induce the non-specific activation of peritoneal macrophages, and strum antibodies including IgE antibody might enhance the cytotoxicity by macrophages,

• The Korean Journal of Mycology
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• v.27 no.2 s.89
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• pp.170-174
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• 1999

Water-soluble polysaccharides from the fruiting bodies of Agaricus blazei Murill were extracted with 0.9% sodium chloride and hot water, successively. The purified polysaccharides showed a potent immunostimulating activity. Eight major polysaccharides, which were named from AG-l to AG-8, were fractionated and purified by ethanol precipitation, ion exchange chromatography on DEAE-cellulose and gel filtration on Toyopearl HW 65F. These polysaccharides were identified to be homogeneous by analysis of HPLC. Three major active polysaccharides (AG-2, -3, and -6) showed relatively strong immunostimulating activity. AG-2 and -3 were composed of glucose, galactose and mannose in the molar ratios of 74.0:15.3:10.7 and 63.6:17.6:12.7, respectively. AG-6 was composed of glucose and ribose in the molar ratios of 81.4:12.6.

• The Korean Journal of Mycology
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• v.18 no.2
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• pp.58-69
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• 1990

ABSTRACT: In order to find physiologically active components from higher fungi, hot-water soluble components were extracted from the basidiocarps of Ganoderma lucidum. The extract was purified and separated by DEAE cellulose ion exchange chromatography and Sepharose CL-4B gel filtration method. The separated fractions were designated CR, IN, IA, GL and GH. Fraction GL showed the highest antitumor activity among the fractions and its molecular weight was found to be 47 KD. The tumor inhibition ratio of Fr. GL was 81 % at the dose of peritoneal administration of 20 mg/kg/day for 10 days in mice. Chemical analysis of this fraction showed 82% polysaccharide, 8% protein and 0.9% hexosamine. The polysaccharide moiety consisted of 63% glucose, 27% galactose, 7% mannose and 3% fucose. Fraction IN was found to increase the amount of superoxide anion in activated macrophages to 1.6-fold and the number of plaques in hemolytic plaque assay to 6-fold, respectively. These results indicate that the antitumor activity was exerted through immunopotentiation, but not through direct cytotoxicity against the tumor.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.250-256
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• 2003

Lactic acid bacteria with amylolytic and acid producing activities can ferment starch directly to lactic acid thereby producing a monomer for the production of biodegradable poly lactic acid (PLA). In this study, the strain producing L-lactic acid from soluble starch was isolated from Nuruk. The isolated strain was identified as Enterococcus sp. through its morphological, cultural, biochemical characteristics as well as the 16S rDNA sequence analysis, and named Enterococcus sp. JA-27. Enterococcus sp. JA-27 produced exclusively L-lactic acid from soluble starch as a carbon source. The optimal conditions for the maximum production of L-lactic acid from Enterococcus sp. JA-27 were 30 C, pH 8, 1.5 % soluble starch as a substrate and 3.5 % tryptone as a nitrogen source, 0.1 % $K_2$$HPO_4$, 0.04 % $MgSO_4$. $7H_2$O, 0.014 % $MnSO_4$$.$4$H_2O$, 0.004% $FeSO_4$$.$$7H_2$O. Batch and fed batch culture were carried out and the former was more effective. L-Lactic acid production in the optimum medium was significantly increased in a 7 L jar fermenter, where the maximum L-lactic acid concentration was 3 g/L. For the purification of lactic acid in fermented broth, two stage ionexchange column chromatographies were employed and finally identified by HPLC.

• Applied Biological Chemistry
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• v.46 no.2
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• pp.79-83
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• 2003

Elicitins, proteinaceous elicitors secreted from Oomycetes fungi (Phytophthora spp. and Pythium spp.), have been known as inducer of hypersensitive response (HR) in incompatible interactions between plant and pathogens. Five elicitins among many Korean Phytophthora species caused the reactions of distal HR in radish, chinese cabbage and some hot pepper cultivars, but not in cucumber and tomato. Because the isolation of elicitin from Phytophthora cambivora hasn't been reported yet, we have purified a cambivorein, a new member of the elicitin family, from the culture filtrate of Phytophtilora cambivora (KACC 40160) by using FPLC (Fast Protein Liquid Chromatography, AKTA) with sepharose S and Sephacryl HR columns. We confirmed that it induces necrosis activities in some hot pepper cultivars and its molecular weight is about 10 KDa by Tricine-SDS-PAGE. Comparison of amino acid sequences of its N-terminal ends also informed the identification of Iysine at the 13th position, which is characteristic of a kind of basic elicitin isoform $({\beta}-isoform)$. It Also showed that our elicitin is not identical with N-terminal sequences of many elicitins reported from Phytophthora spp..

• Korean Chemical Engineering Research
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• v.49 no.2
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• pp.129-136
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• 2011

Simulated Moving Bed(SMB) process consists of multiple chromatographic columns, which are usually partitioned into four zones. Such a process characteristic allows a continuous binary separations those are impracticable in conventional batch chromatographic processes. Compared with batch chromatography, SMB has advantages of continuity, high purity and productivity. Various researches have been reported for the integration of reaction and recovery during process operation on the purpose of economics and effectiveness. Simulated Moving Bed Reactor(SMBR) is introduced to combine SMB as a continuous separation process and reactor. Several cases of SMBR have been reported for diverse reactions with catalytic, enzymatic and chemical reaction on ion exchange resin as main streams. With an early type of fixed bed using catalyst, SMBR has been developed as SMB using fluidized enzyme, SMB with immobilized enzyme and SMB with discrete reaction region. For simple modeling and optimization of SMBR, a method considering convection only is possible. A complex method considering axial dispersion and mass transfer resistance is needed to explain the real behavior of solutes in SMBR. By combining reaction and separation, SMBR has benefits of lower installation cost by minimizing equipment use, higher purity and yield by avoiding the equilibrium restriction in case of reversible reaction.

• The Korean Journal of Food And Nutrition
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• v.24 no.3
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• pp.362-366
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• 2011

The objective of this study was to compare the anticancer activity of glucosinolates against four different cancer cells; SNU-251, SNU-354, SNU-C4, MCF-7 and to determine the amounts of glucosinolates in mustard leaves. Green and red mustard leaves were cultivated on the field of Rural Development Administration from May to July, 2010. After the leaves were harvest and lypophilized, the fine powder was extracted with boiling 70% methanol(v/v) for the measurement of anticancer activity and then applied to the cancer cells obtained from Korean Cell Line Bank(KCLB). The anticancer activity of red leaf mustard was higher than that of green mustard leaf, particularly in SNU-251 for 24 hr and in SNU-C4 for 48 hr. In both green and red mustard leaf, sinigrin was the most abundant glucosinolte and was determined as 4.71 and 3.06 mg/g dry wt., respectively, but glucoiberin only presented 0.14 mg/g dry wt. with minor amounts.

• Korean Journal of Food Science and Technology
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• v.28 no.4
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• pp.684-688
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• 1996

In order to search for antithrombotic peptides from soybean paste, the inhibitory activity of water extract of soybean paste and its peptide fractions on ADP-induced aggregation of washed platelets was assayed. Soybean paste extract treated with ultrafiltration (M.W. cut off, 3,000 daltons) was found to have inhibitory activity of 90% at the dose of $96\;{\mu}g/ml$ by the method of turbidometric aggregometer. Soybean paste extract was fractionated to 19 fractions (No. B-18) by Dowex 50W X-2 ion exchange column chromatography and activity test was performed by the microplate method. All of the fractions had antithrombotic activity $(IC_{50},\;101,000\;{\mu}g/ml)$, and most fractions had higher activity than positive control, RGDS $(IC_{50}.\;205\;{\mu}g/ml)$. Especially, basic fractions No.16-18 showed higher activity than soybean paste extract $(IC_{50},\;30\;{\mu}g/ml)$. The fraction No.16 with the highest activity $(IC_{50},\;10\;{\mu}g/ml)$ was purified and analysed for amino acid composition. The results showed that histidine, arginine, and alanine were major residues in the peptide part of the fraction.

• The Korean Journal of Food And Nutrition
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• v.15 no.2
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• pp.104-111
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• 2002

The peptidyl prolyl sis-trans isomerase (PPIase, EC 5.2.1.8) from bacillus stearothermophilus was extracted from the cells treated with by lysozyme. PPIase was purified from the cell extracts by heat treatment, ammonium sulfate precipitation, ion exchange chromatography and finally gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The molecular weight of the purified PPIase was estimated as 18kDa by SDS-PAGE. The 39 amino acid residues from the N-terminus were determined by the protein sequencer. The enzyme showed the optimum pH at 8.0 and was stable at the range of pH 7.0∼8.0. The enzyme was considerably stable after heat treatment at 60$\^{C}$ for 30minutes, and the enzyme was quite stable up to 65$\^{C}$. The presence of the PPIase in the refolding solution accelerated the isomerization rate of the assay peptide. PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-l primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPI-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus.