• Archives of Pharmacal Research
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• v.21 no.5
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• pp.555-558
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• 1998

chondroitin sulfates were isolated from the mud snail. For the quantitative analysis of enzymatic digestion products of isolated chondroitin sulfates, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. by the action of chondroitinase ABC, three unsaturated disaccharides$ 2-acetamide-2-deoxy-3-O-({\beta}-D-gluco-4-enepyranosyluronic acid)-D-galactose $$({\Delta}Di-OS), $2-acetamide-2-deoxy-3-O-({\beta}-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose ({\Delta}Di-6S) and 2-acetamide-2-deoxy-3-O-({\beta}-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose ({\Delta}Di-4S)$ were produced from the mud snail chondroitin sulfates. The analysis showed that relative proportion of ${\Delta}Di-OS/{\Delta}Di-6S/{\Delta}Di-4S$ was 58.7/3.1/38.2. The immunomodulating activity of chondroitin sulfate was examined by cell proliferation assay and these results suggest that it might be a immunosuppressant.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.4
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• pp.641-645
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• 2004

Crude chondroitin sulfates extracted from midduck tunics (Styela clava) and munggae tunics (Halocynthia roretzi) were examined in vivo in order to be utilized as a cosmetic material which was followed by an in vitro assay. Examinations, such as acute oral toxicity, skin sensitization, acute eye irritation, and primary skin irritation, were peformed with a variety of laboratory animals. Phototoxic and photosensitization tests were not conducted since all chondroitin sulfates failed to absorb U.V. light at the range of 280 to 420 nm. In acute dermal and eye irritation, both specific clinical signs and dead cases were not demonstrated during the test period, but crude chondroitin sulfates from midduck and munggae tunics, and standard chondroitin sulfate from bovine trachea were showed 2.5, 1 and 1.25 of acute ocular irritation index (A.O.I.), respectively. In the case of skin sensitization, crude chondroitin sulfate from midduck tunics exhibited neither specific clinical signs nor dead cases in the entire course of the examination. While in acute oral toxicity, crude chondroitin sulfates from both midduck and munggae tunics found neither specific clinical signs nor dead cases during the test, and LD50 was suspected to be over 2 g/kg. Based on this study, it was proven that crude chondroitin sulfates from either midduck or munggae tunics can be used safely as a cosmetic material.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.4
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• pp.646-652
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• 2004

With the aim of using a cosmetic material, chondroitin sulfates extracted from midduck tunics (Styela clava) and munggae tunics (Halocynthia roretzi) were examined in vitro with two cell lines for cell toxicity, collagen synthesis, cell growth and recovery ability after U.V. irradiation. Cell toxicity test with A 431 and CCD 1108Sk was able to observe high activity between 400 and 600 $\mu\textrm{g}$/m while standard chondroitin sulfate (CS) purchased from Sigma was showed at 80 $\mu\textrm{g}$/mL. Even fraction 1 and 2 collected from chondroitin sulfates originated from midduck appeared having the highest activity between 600 and 1000 $\mu\textrm{g}$/mL, but slightly lower compared to crude chondroitin sulfates from both mideduck and munggae. In cell growth examination, it was not able to find significant differences between chondroitin sulfates used. Both crude chondroitin sulfates were exhibited the highest activity for two cell lines except that of mideduck which was showed activity for CCD 1108Sk. CS, fraction 1 and 2 from midduck were not able to demonstrate a significant activity in collagen synthesis. On the contrary, crude chondroitin sulfates from both munggae and midduck were showed the highest activity at 100 and 50 $\mu\textrm{g}$/mL with only CCD 1108Sk. The recovery ability after U.V. irradiation with crude chondroitin sulfates from both munggae and midduck were showed high activity at 400 $\mu\textrm{g}$/mL with CCD 1108Sk and A 431. But there were no activity observed in fractions examined, As a consequence, the crude chondroitin sulfates from both munggae and midduck might not only be available as a cosmetic material but also useful for increasing some activity by blending properly.

• Archives of Pharmacal Research
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• v.23 no.2
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• pp.182-186
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• 2000

Chondroitin sulfates proteoglycans were isolated from human placenta. For the identification of enzymatic digestion products of isolated proteoglycan, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitin ABC and chondroitin B lyase, three unsaturated disaccharides 2-acetamide-2-deoxy-3-O-($\beta$-D-gluco-4-enepyranosyluronic acid)-D-galactose ($\delta$Di-OS), 2-acetamide-2-deoxy-3-O-($\beta$-D-gluco-4-enepyranosyluronic acid)-6-O-su lfo-D-galactose ($\delta$Di-6S) and 2-acetamide-2-deoxy-3-O-($\beta$-D-gl uco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose ($\delta$Di-4S) were produced from the human placenta proteoglycan. The anticoagulant activity of chondroitin sulfate proteoglycan was evaluated by activated partial thromboplastin time (aPTT) assay and thrombin time (TT) assay. The clotting times of aPTT and TT were increased from 72 to 144 sec and 19 to 27 sec, respectively. The Immune-modulating activity of chondroitin sulfate proteoglycan was examined by cell proliferation assay and these results suggest that it may play a role in suppression of the function of immune-related cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.2
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• pp.350-358
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• 1998

The antimutagenic and anticancer activities of glycoprotein(GP) and chondroitin sulfate(CS) from sea cucumbers were studied using Ames mutagenicity test and human cancer cells culture test. The GP's inhibitory effect toward aflatoxin B1(AFB1) and 3, 2'-dimethyl-4-amino-biphenyl(DMAB) increased with the higher added concentrations up to 5% level(w/w) regardless fractionation methods. The GP from sea cucumbers through DEAE-cellulose column chromatography showed an inhibitory effect ranged from 84 to 98%, and the maximum antimutagenicities resulted in red sea cucumber with 98% (AFB1) and 95% (DMAB). But 5% level of CS from various sea cucumbers had an inhibitory effect toward those both indirect mutagens ranged from 79% to 85%. However, in case of direct mutagens(N-methyl-N'-nitro-N-nitrosoguanidine, MNNG and 4-nitroquinoline-1-oxide, 4-NQO), the GP's inhibitory effect was 55∼78% and the CS had a low inhibitory effect(58∼70%) at the added level of 5%. The GP from sea cucumbers exhibited the strong inhibitory effects with 89∼95% and 82∼92% on the growth of HT-29 human crcinoma cells and AZ-521 human gastric cancer cells (at 5% level).

• Fisheries and Aquatic Sciences
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• v.13 no.3
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• pp.210-215
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• 2010

In an effort to isolate a bacterium producing chondroitin sulfate (CS), a marine bacterium, KS-1, which produces mucopolysaccharides, was isolated from seawater and identified as Marinobacter sp. based on analyses of its morphological and biochemical traits and 16S rDNA sequence. Agarose-gel electrophoresis showed that the KS-1 strain produces a CS-like mucopolysaccharide. Structural analysis using Fourier transform infrared spectroscopy revealed that the structure of the CS-like mucopolysaccharide produced by Marinobacter sp. KS-1 is similar to that of dermatan sulfate (CS B). However, the molecular mass of the CS-like mucopolysaccharide is higher than that of standard chondroitin sulfates. Considering the above results, we conclude that the Marinobacter sp. KS-1 produces a CS-like mucopolysaccharide that differs somewhat from CS B in molecular mass.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.1
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• pp.72-80
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• 1997

To elucidate food value and medicinal effect of sea cucumbers, sugar composition of those gly-coprotein and chondroitin sulfate was studied. The contents of sulfate esters in sea cucumbers were 1.21%(blue), 0.90%(red) and 1.19%(black). Predominant carbohydrates were identified as fucose, glucose, D-mannuronic acid and N-acetylglucosamine, and those amount was more than 80% to total carbo-hydrate, while the minor sugar composition was ribose, mannose, galactose, N-acetylgalactosamine and D-glucuronic acid. Also, the major carbohydrate moiety of glycoproteins of sea cucumbers was revealed as fucose, mannose, N-acetylglucosamine, glucose and ribose, and those amount was more than 86% to total carbohydrate. Galactose, N-acetylgalactosamine, D-glucuronic acid and mannuronic acid were minor carbohydrate moiety. The contents of sulfate esters in glycoproteins were 0.96% for blue sea cucumber, 1.15% for red sea cucumber and 1.13% for black sea cucumber, while those in chondroitin sulfates were 3.52%(blue), 3.60%(red) and 3.72%(black). The carbohydrate moiety of chondroitin sulfate was identified as N-acetylgalactosamine (73~ 87%), fucose (7~15%) and D-glucuronic acid(5~12%). As the base on the IR spectrum of strong absorption appeared in 1240$cm^{-1}$ / for stretching vibrations in S=0 group and weak absorptions in 850$cm^{-1}$ / and 820$cm^{-1}$ /for stretching vibrations in C-0-S group, chondroitin sulfates had sulfate group which was bound to $C_4$in fucose.

• Proceedings of the PSK Conference
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• 2000.04a
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• pp.144.1-144.1
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• 2000

• Food Science and Biotechnology
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• v.18 no.4
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• pp.872-877
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• 2009

By-products of marine organisms including salmon, skate, flatfish, and yellow goosefish were investigated to search for new source of chondroitin sulfate (CS). Agarose gel electrophoresis with chondroitinase depolymerization showed that purified chondroitin sulfate did not contain any other glycosaminoglycans. 1H-nuclear magnetic resonance (NMR) spectra were acquired to confirm the structure and purity. The average molecular weight ranging from 22 to 64 kDa was determined by high performance size exclusion chromatography. Disaccharide compositions and purities were determined by strong anion exchange-high performance liquid chromatography (SAX-HPLC) after chondroitinase ABC depolymerization. SAX-HPLC data exhibited that the purity was from $81.7{\pm}1.3$ to $114.2{\pm}2.5%$ and the yield was from 1.3 to 12.5%. All analytical results indicate that salmon cartilage, skate cartilage, and yellow goosefish bone could be promising sources of CS to substitute shark cartilage CS in commercial neutraceuticals.

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.140-144
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• 2016

The gene encoding chondroitinase ABC from a genomic library of Bacteroides stercoris HJ-15, which was isolated from human feces, was cloned. The cloned gene consisted of 3,090 bp and was predicted to encode a 1,029−amino-acid protein. The B. stercoris chondroitinase ABC gene was not homologous to other chondroitinase ABC genes; however, its amino acid sequence showed 71% homology to that of Bacteroides thetaiotaomicron. The gene was cloned in the pET-26b+ expression vector and expressed under the T7 promoter in Escherichia coli BL21(DE3). The purified recombinant chondroitinase ABC degraded chondroitin sulfates A, B, and C.