Shewanella oneidensis PKA 1008의 조효소를 이용한 경단구슬모자반 (Sargassum muticum) 및 쌍발이모자반 (Sargassum patens C. Agardh) 추출 분해물의 특성 및 항염증 효과

Abstract

Brown algae, such as Sargassum muticum (SM) and SargassumpatensC.Agardh (SP), contain 30 to 60% carbohydrates and polysaccharides such as alginate, fucoidan, laminararan and mannitol, etc. Moisture, ash, crude lipid, crude protein, total sugar and alginate contents of SM were 12.30%, 19.01%, 1.65%, 12.20%, 53.35% and 26.46%, and SP were 8.29%, 21.64%, 0.60%, 18.10%, 51.37% and 32.37%, respectively. This study was conducted to investigate the physicochemical properties and anti-inflammatoryeffect of the enzymatic extracts of SM and SP using crude enzyme from Shewanella oneidensis PKA 1008. S. oneidensis PKA 1008 strain, producing an alginate-degrading enzyme, was isolated from Ulva pertusa. To obtain the optimal degrading conditions for the SM and SP, the SM and SP were prepared in a ratio of 1:1 (v/v) with crude enzyme from S. oneidensis PKA 1008, respectively. The mixtures were reacted at 30°C for 0 to 60 h. Changes in pH, color value, reducing sugar, viscosity and thin-layer chromatography (TLC) of SM and SP degrading extracts were measured. The reducing sugar of SM increased significantly as the incubation time increased, and showed the highest reducing sugar is 474.35 μg/mL at 48 h. SP showed the highest reducing sugar is 571.91 μg/mL at 24 h. The pH and viscosity were significantly reduced at 60 h compared to 0 h. The degraded alginate oligosaccharides were visualized by TLC developed with a solvent system of n-butanol:methanol:water, 4:1:2 (v/v). Alginate were degraded into dimmers at 60 h. In the LPS-stimulated macrophage RAW 264.7 cells, the secretion of cytokines such as TNF-α, IL-6, IL-1β was inhibited by the treatment of SM and SP enzymatic extracts. These results indicate that crude enzyme from S. oneidensis PKA 1008 can be used to improve the polysaccharide degradation of SM and SP.