• Applied Biological Chemistry
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• v.22 no.1
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• pp.1-9
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• 1979

The physical and chemical properties of ${\gamma}-conglycinin$ of soybean (Glycine max) were investigated. The soybean protein extracted from soybean meal using 0.2M NaCl solution at pH 4.5 was passed through a Sephadex G-150 column to isolate 7S globulin. ${\gamma}$-Conglycinin was isolated and purified from the 7S globulin with a DEAE Sephadex A-50 column chromatography. The protein preparation was pure on immunoelectrophoresis, polyacrylamide gel electrophoresis and gel isoelectric fouling. It had an isoelectric point at pH 5.4 and contained 16.12% nitrogen, 4.18% mannose and 1.21% glucosamine. Amino acid composition, in general, shaved that ${\gamma}-conglycinin$ contained higher contents of lysine, dicarboxylic acids and ammonia nitrogen, and lower contents of sulfur-containing amino acids and tryptophan. The subunits of ${\gamma}-conglycinin$ were distributed in the range of pH 4.6-5.5. The subunits located in the pH region of 4.6-5.0 and 5.0-5.5 were glycopeptides (molecular weight of 38,000) and simple peptide (MW of 32,000), respectively.

• Applied Biological Chemistry
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• v.35 no.6
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• pp.462-469
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• 1992

Two kinds of complement activating (anti-complementary) polysaccharides, which were expected to be immunomodulators were purified from Arecae Pericarpium (the pericarps of Areca catechu), and their action modes have been studied. The active polysaccharides, AC-2-IIIa and AC-2-IIIc from Arecae Pericarpium showed dose-dependent anti-complementary activities on $TCH_{50}$. The anti-complementary activities of AC-2-IIIa and AC-2-IIIc in metal ion-free condition were completely decreased in comparison with control whereas in case of $Ca^{2+}$-free condition, these activities were maintained, considerably. Also AC-2-IIIa and AC-2-IIIc showed relatively potent alternative complement pathway activities. Furthermore, after incubation of the normal human serum with polysaccharide of Arecae Pericarpium in the absence of $Ca^{2+}$ ion, a cleavage of C3 in the serum was found to have occurred through immunoelectrophoresis (IEP) with anti-human C3. Also, from the results of IEP using anti-human whole serum, the ratios of the height of 3rd peak to ${\alpha}2-M$ peak by AC-2-IIIa and AC-2-IIIc proved to be $1.50{\pm}0.04$ and $1.22{\pm}0.08$, respectively. These results indicate that the modes of complement activation by AC-2-IIIa and AC-2-IIIc from Arecae Pericarpium are via both the classical pathway and the alternative pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.63-69
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• 2011

To characterize the polysaccharides which exist as soluble forms in Korean traditional alcoholic beverages, the polysaccharides were isolated from Korean pear wine and their anti-complementary activities were examined. The main polysaccharide, PW-1 was purified to homogeneity from the crude polysaccharide (PW-0) in pear wine by size exclusion chromatography using Sephadex G-75. Molecular mass of PW-1 was estimated to be 150 kDa and it contained significant proportion of mannose (81.8%) and 5 different minor component sugars such as arabinose (1.2%), galactose (2.7%), glucose (8.5%), galacturonic acid (5.3%) and glucuronic acid (0.5%). These analyses indicated that the main polysaccharide in pear wine was mainly present as a mannan which had originated from the cell walls of fermenting yeasts. On the other hand, PW-1 showed potent anti-complementary activity in a dose-dependent fashion. Identification of C3 activation products by the crossed immunoelectrophoresis using anti-human C3 and anti-complementary activity of PW-1 in $Ca^{++}$-free condition suggested complement activations by PW-1 from Korean pear wine occur via both classical and alternative pathways.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.1
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• pp.95-102
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• 2012

To examine the new practical utilization of mucilages in Opuntia humifusa, the Korean pear cactus, the polysaccharides were isolated from O. humifusa, and immuno-stimulating activities were assayed. The main polysaccharide, CNC-E, was prepared by a commercial enzyme treatment, water extraction, and ethanol precipitation. The molecular mass of CNC-E was estimated to be about 700 kDa, and it consisted mainly of arabinose, galactose and xylose in addition to two minor sugars such as rhamnose and fucose. On the other hand, CNC-E showed considerably high splenocyte proliferation activity in a dose-dependent manner. Peritoneal macrophages stimulated with CNC-E produced cytokines such as IL-6, IL-12, IL-10, and TNF-${\alpha}$. The intravenous administration of CNC-E significantly augmented the cytotoxicity of natural killer (NK) cells against Yac-1 tumor cells. Especially, NK cells obtained from the mice treated with $100{\mu}g$ of CNC-E showed threefold higher cytolytic activity than those of untreated mice. CNC-E also showed potent anti-complementary activity in a dose-dependent fashion. Identification of C3 activation products by the crossed immunoelectrophoresis using anti-human C3 and the anti-complementary activity of CNC-E in a $Ca^{2+}$-free condition suggested complement activations by CNC-E that occur via both alternative and classical pathways. These results indicate that Korean pear cactus contains selected polysaccharides that provide immuno-stimulating activities beneficial to human health.

• Applied Biological Chemistry
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• v.35 no.1
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• pp.42-50
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• 1992

We conducted screening on Chinese medicinal herbs to examine their anti-complementary activity by hemolytic complementary assay $(TCH_{50})$. Among 55 kinds of herbs, several herbs showed relatively potent anti-complementary activity which decreased $TCH_{50}$, more than 70% in comparison with control. Then, hot water extracts of the following herbs, Curcuma aromatica, Areca catechu, Gleditsiae spina, Euonymus alata, Acanthopanax senticous. Lonicera japonica, Aconitum carmichaeli, Curcuma zedoaria and Cinnamoum cassia, which were shown relatively potent anti-complementary activity were partially purified and analyzed their chemical properties. These activities were resistant to digestion with pronase but decreased by treatment with $NaIO_4$. These results may indicate that the complement activating ability in their herbs is due to polysaccharide. Furthermore, the anti-complementary activity of Areca catechu which was showed the most potent activity, was reduced partially in the absence of the $Ca^{++}\;ion$. After incubation of the normal human serum with partially purified polysaccharide of A. catechu in the absence of $Ca^{++}\;ion$, a cleavage of C3 in the serum was found to have occurred through immunoelectrophoresis using rabbit anti-human C3 serum. These results indicate that the mode of complement activation by polysaccharide of A. catechu is via both the alternative and classical pathway.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.290-295
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• 2002

Among 12 lactic acid bacteria examined for their abilities to activate the complement system by hemolytic complement assay $(TCH_{50})$, Lactobacillus plantarum previously isolated from Kimchi showed high anti-complementary activity. The anti-complementary activity of the cell wall fraction of L. plantarum was more potent than that of the cytosol fraction, and both activities showed dose dependency. These high activities of the cytosol and the cell wall fractions were relatively resistant to the digestion with pronase, but sharply decreased after the treatment of $NaIO_4$. These results suggested that the complement activation by the cytosol and the cell wall fractions was mainly due to their polysaccharides. By the cross-immunoelectrophoresis using anti-human C3, the C3 activation products from both fractions were identified in $Ca^{++}$-free condition. Anti-complementary activity $(ITCH_{50})$ of the cell wall fraction was retained under the same condition, whereas that of the cytosol fraction was reduced considerably. From these results, it was inferred that the mode of complement activation by the cell wall fraction was mainly via alternative pathway, and that of the cytosol fraction was via both alternative and classical pathways.

• Korean Journal of Food Science and Technology
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• v.25 no.3
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• pp.197-203
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• 1993

Screenings were performed on edible plants to examine their complement-system activating ability (anti-complementary activity) by hemolytic complement assay $(TCH_{50})$. Among 38 kinds of plant extracts, 5 kinds showed relatively strong anti-complementary activity which decreased $TCH_{50}$ more than 60% comparison with control and the order of activity was Zingiber officinale>Colocasia antiquorum>Capsella bursapastoris>Ginkgo biloba>Alium monanthum in $1000{\mu}g/ml$. The anti-complementary activity of ZR-1 prepared from the root of Zingiber officinale which was showed the most potent activity, did not change by pronase treatment, but decreased greatly by periodate oxidation. These results indicate that not protein moiety but carbohydrate moiety in ZR-1 fraction may also contribute to the anti-complementary activity. Also, the anti-complementary activity of ZR-1 was reduced partially in the absence of the $Ca^{2+}$ ion. When crossed immunoelectrophoresis using anti-human C3 serum was carried out after incubation of normal human serum with the ZR-1 in $Ca^{2+}$ free condition, a cleavage of C3 precipitin line was observed. Furthermore this polysaccharide fraction considerably inhibited $ACH_{50}$. These results also indicate that the mode of complement activation by polysaccharide from Zingiber officinale is via not only the classical pathway but also the alternative pathway.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.879-884
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• 2002

Cold and hot water fractions of Diospyros kaki were screened to determine its anti-complementary activity. Flour of Diospyros kaki leaf (250 g) was boiled at $100^{\circ}C$ for 3 h and passed through a membrane of 10 kDa molecular weight (DK-0). DK-0 was precipitated with ethanol and refluxed with methanol to obtain the crude polysaccharide (DKC). DKC-1 was isolated by ion exchange chromatography on DEAE-Toyopearl 650C, and DKC-1c was purified from DKC-1 by size exclusion chromatography on Bio gel P-60. The anti-complementary activities of DKC-1c at $1000\;{\mu}g/mL$ were 85.4 and 61.1% via whole and alternative pathways, respectively. DKC-1c was determined as a neutral polysaccharide composed of glucose (29.0 mol.%), arabinose (24.3 mol.%), and galactose (16.2 mol.%) with the molecular weight of 66.6 kDa. Results of agarose gel immunoelectrophoresis revealed DKC-1c, as a complement activator, cleaved C3 into C3a and C3b via both pathways.

• Journal of Aquaculture
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• v.8 no.1
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• pp.1-19
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• 1995

In red seabream, Pagrus major the female specific protein in the vitellogenic female serum was identified by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The female specific serum protein might be vitellogenin based on the results of the immunological analysis for the male and vitellogenic female sera and crude egg extracts. Also, it was identified by the immunodiffusion test that the purified yolk protein from ovarian egg extracts has antigenic identities shared with the female specific serum protein. To study the relationship between the maturational stages of gonad and plasma levels of vitellogenin, these were measured from the late resting period (January) to the vitellogenic preiod (April) by the modified enzymeimmunoassay (EIA) using antiserum against yolk protein. The level of plasma vitellogenin began to increase in February (previtellogenesis stage) and continuously increased with the ovarian growth during the vitellogenesis period (March to April). The plasma vitellogenin levels were significantly different between the females and the males in February. Validation for the modified EIA system. was tested .The absorbance curve of serial dilutions of serum from the vitellogenic female was paralleled to the standard curve of yolk protein; $109\pm5.6\%$ recovery was achieved by the modified EIA. And the intraassay coefficients of variation were less than 10% within the concentration ranging from 31.3 ng/ml to 1,000 ng/ml. These findings suggest that the sex determination in adult red seabreams could be possible by using the modified EIA as early as in February.

• Korean Journal of Food Science and Technology
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• v.40 no.2
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• pp.220-227
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• 2008

In this study, polysaccharides were isolated from Korean persimmon vinegar to characterize the polysaccharides existing as soluble forms within traditional Korean fermented beverages, and their immuno-stimulating activities were examined. Three successive chromatographies were used to purify the main polysaccharide in the persimmon vinegar, PV-1b-I, to homogeneity from the crude polysaccharide (PV-0). The molecular mass of PV-1b-I was estimated as 110 kDa and it contained significant proportions of mannose (46.8%), galactose (28.5%) and arabinose (19.1%). PV-1b-I strongly reacted with ${\beta}$-glucosyl Yariv reagent, suggesting the presence of an arabino-3,6-galactan moiety. PV-1b-I also induced high levels of macrophage activation and mitogenicity on murine splenocytes in vitro. The intravenous administration of PV-1b-I significantly augmented NK cytotoxicity against YAC-1 tumor cells. PV-1b-I also showed potent anticomplementary activity in a dose-dependent manner. Finally, C3 activation products were identified by crossed immunoelectrophoresis using anti-human C3 and the anti-complementary activity of PV-1b-I under $Ca^{2+}$-free conditions, suggesting that this PV-1b-I causes complementary activations via both alternative and classical pathways. From these results, one can conclude that Korean persimmon vinegar contains select polysaccharides in addition to healthy components, and these polysaccharides appear to provide immuno-stimulating activities beneficial to human health.