Rafiquzzaman S M, Kim Eun-Young, Kim Yu-Ri, Nam Taek-Jeong, Kong In-Soo
Department of Biotechnology, Pukyong National University, Busan 608-737, Republic of Korea.
Int J Biol Macromol. 2013 Nov;62:265-72. doi: 10.1016/j.ijbiomac.2013.09.009. Epub 2013 Sep 20.
The present study was performed to investigate the chemical composition and antioxidant activity of glycoprotein purified from Undaria pinnatifida Harvey (UPGP). On SDS-PAGE, UPGP migrated as a single band with a molecular weight of approximately 10 kDa and confirmed by staining with Schiff's reagent as glycoprotein. It consists of a carbohydrate component (42.53%) and protein component (57.47%). Amino acid profile, FT-IR spectrum and enzymatic glycosylation analysis suggested that protein is linked with carbohydrate by O-glycosylation. UPGP showed dose-dependent antioxidant activities as detected by different assays before and after in vitro digestion. The IC50 values of undigested UPGP were 0.25 ± 0.03, 0.08 ± 0.005, 0.69 ± 0.12, and 0.25 ± 0.08 mg/mL for DPPH, ABTS, FRAP, and NO, respectively. Following in vitro digestion, the antioxidant activities of UPGP were decreased during the gastric phase compared to those of undigested UPGP, with an increase occurring during the duodenal phase in all assays. However, the reducing power was unchanged after in vitro digestion. Furthermore, UPGP showed protective activity against oxidative DNA damage both undigested, after saliva and duodenal phase of digestion. These results indicate that the antioxidant and DNA protection activities of UPGP may be pH-dependent and assay specific.
本研究旨在探究从海带(Undaria pinnatifida Harvey,UP)中纯化得到的糖蛋白(UPGP)的化学成分和抗氧化活性。在SDS-PAGE电泳中,UPGP迁移为一条单一的条带,分子量约为10 kDa,经席夫试剂染色确认为糖蛋白。它由碳水化合物成分(42.53%)和蛋白质成分(57.47%)组成。氨基酸谱、傅里叶变换红外光谱和酶促糖基化分析表明,蛋白质通过O-糖基化与碳水化合物相连。体外消化前后,通过不同检测方法检测发现,UPGP呈现出剂量依赖性的抗氧化活性。未消化的UPGP对DPPH、ABTS、FRAP和NO的IC50值分别为0.25±0.03、0.08±0.005、0.69±0.12和0.25±0.08 mg/mL。体外消化后,与未消化的UPGP相比,UPGP在胃消化阶段的抗氧化活性降低,而在所有检测中十二指肠消化阶段活性增加。然而,体外消化后其还原能力不变。此外,无论是未消化的UPGP,还是经过唾液和十二指肠消化阶段后的UPGP,均对氧化DNA损伤具有保护活性。这些结果表明,UPGP的抗氧化和DNA保护活性可能依赖于pH值且具有检测特异性。