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Article Title

Isolation and analysis of a novel protoglycan from zizyphus jujuba cv. jinsixiaozao

Abstract

Chinese jujube (Zizyphus jujuba Miller) is a well-known oriental medicinal plant that has various biological activities. In this study, a novel water-soluble proteoglycan (ZSG4b) was isolated from Zizyphus jujuba cv. Jinsixiaozao and its effect on the complement system was investigated. ZSG4b was extracted with hot water and purified by DEAE-Sepharose CL-6B and Sepharose CL-6B column chromatography. It was eluted as a single symmetrical narrow peak on high-performance gel-permeation chromatography (HPGPC) and the average molecular weight was estimated to be 1.4 × 105 Da. ZSG4b contained 83.5% glycan and 9.7% protein. Gas chromatography analysis result indicated that the glycan portion of ZSG4b was composed mainly of rhamnose, arabinose, mannose and galactose in a molar ratio of 13.8:4:3:8. It also contains 29% uronic acid as determined by m-hydroxybiphenyl method. The presence of galacturonic acid was estimated by the increase of galactose content in the carboxyl-reduced ZSG4b. The amino acid composition showed that it was rich in aspartic acid, glutamic acid, serine, threonine and leucine; the molar content of these five amino acids constituted 62.8% of the total amino acids. The β-elimination reaction demonstrated that the protein and glycan were linked by O-linkage. ZSG4b exhibited high anti-complement activity which was increased in accordance with the concentration below 120 μg/mL in vitro.

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