Analysis of the polysaccharide fractions isolated from pea (Pisum sativum L.) at different levels of purification

Crude pea (Pisum sativum L.) polysaccharides (CPPs) were extracted under ultrasound assistance, and CPP yield was highest to 6.27381%, which optimized using response surface methodology. Enzymatic method was more effective in deproteinization than Trichloroacetic acid and Sevag method, when considering the polysaccharide retention value as well as the protein clearance. Three-phase partitioning deproteinization indicated that the combination of the enzyme and Sevag method was more effective than their single use. Pea polysaccharide fractions were obtained by diethylaminoethyl-52 cellulose (W-DE-PP, N-DE-PP1, and N-DE-PP2) and Sephadex G-100 size-exclusion chromatography (W-DE-GPP, N1-DE-GPPa, and N1-DE-GPPb) in that order. Polysaccharide fractions W-DE-GPP and N1-DE-GPPa were showed a smooth surface with many cavities by Scanning electron microscopy (SEM) in 1,000 folds. All polysaccharides, characterized by high-performance liquid chromatography (HPLC), were composed of rhamnose, arabinose, galactose, glucose, and mannose, with the highest concentrations of galactose and glucose. Compared with different purification levels, N-DE-GPP showed the strongest activity against 2,2-diphenyl-1-picrylhydrazyl and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) free radicals and the highest ferric reducing antioxidant power, which were similar to the results of W-DE-GPP. Therefore, W-DE-GPP and N-DE-GPP may be promising natural sources of antioxidants. PRACTICAL APPLICATIONS: Recently, numerous studies on the extraction, purification, characteristics, and bioactivities of polysaccharides have been conducted. We mainly focused on the functional compounds of legumes. Comprehensive studies on pea polysaccharides are limited. Therefore, in the present study, extraction of CPPs was performed to optimize conditions using response surface methodology. Polysaccharide fractions were obtained from different purification levels and were chemically characterized using HPLC and SEM. Antioxidant activities of polysaccharides with different purification levels were determined. All the conventional methods, described in previous studies, were applied in the study. Furthermore, we analyzed and compared the characteristics of polysaccharides at different purification levels. We believe that our results would likely supplement the fundamental studies on pea polysaccharides.

Keywords: antioxidant activity; characterization; isolation; pea; polysaccharides; purification levels.