• 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.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.347-356
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• 2019

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods. B. velezensis BS2 grows well at 15% NaCl and at $10^{\circ}C$. When B. velezensis BS2 was cultivated in TSB broth for 96 h at $37^{\circ}C$, the culture showed the highest fibrinolytic activity ($131.15mU/{\mu}l$) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and $37^{\circ}C$, respectively. $K_m$ and $V_{max}$ were 0.15 mM and $39.68{\mu}M/l/min$, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong ${\alpha}$-fibrinogenase and moderate ${\beta}$-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at $10^{\circ}C$, B. velezensis BS2 can be used as a starter for jeotgal.