刺五加新品种‘紫加1号'多糖分离纯化及抗氧化活性分析

Isolation and purification the polysaccharides and its antioxidant activity of ‘Zijia 1' novel variety of Acanthopanax senticosus

关键词:  紫加1号, 嫩茎叶, 多糖, 分离纯化, 单糖组成, 分子量, 抗氧化活性

Key words:  ‘Zijia 1', tender stems and leaves, polysaccharides, isolation and purification,monosaccharide compositions, molecular weight, antioxidant activity

摘要:

‘紫加1号'为该课题组选育的刺五加(Acanthopanax senticosus)新品种,嫩茎叶灰紫色且味甘。为分离纯化‘紫加1号'嫩茎叶多糖,并测定各组分的单糖组成和分子量大小、评价各组分的抗氧化活性。该研究以‘紫加1号'嫩茎叶为材料,采用水提醇沉工艺,经大孔树脂吸附法除杂脱色得到粗多糖(A. polysaccharides,ASPS); 通过DEAE-Cellulose 52离子柱和Sephadex G-100凝胶柱分离纯化得多糖组分; 采用离子色谱和凝胶色谱-示差-多角度激光光散射色谱,测定各均一组分的单糖组成和分子量; 通过测定均一组分清除羟基自由基(·OH)、超氧阴离子自由基(O₂^-·)、1,1-二苯基-2-三硝基苯肼自由基(DPPH)的能力,分析其体外抗氧化活性强弱。结果表明:ASPS经分离纯化得到4种多糖组分,即酸性多糖ASPA-1-1、ASPA-2-1、ASPA-3-1和中性多糖ASPN-1,其分子量依次为8.10、26.15、0.91、0.89 kDa,主要是由阿拉伯糖、鼠李糖、半乳糖、葡萄糖、木糖、甘露糖、核糖、半乳糖醛酸和葡萄糖醛酸等按不同比例组成的杂多糖。ASPA-1-1、ASPA-2-1、ASPA-3-1、ASPN-1均具有明显的抗氧化活性,其中ASPA-2-1清除·OH、DPPH的能力高于ASPA-1-1、ASPA-3-1、ASPN-1,而ASPA-3-1清除O₂^-·的能力最强。综上认为,从‘紫加1号'中分离纯化得到的多糖具有较好的抗氧化活性。该研究结果为‘紫加1号'作为天然抗氧化剂的深度研究和进一步开发与利用提供了一定的科学理论依据。

Abstract:

‘Zijia 1' is a new variety of Acanthopanax senticosus bred by our team, its tender stems and leaves characterized with gray-purple color and sweet taste. This study is aim to isolate and purify the polysaccharides from the tender stems and leaves of ‘Zijia 1', and determine the monosaccharide composition and molecular weight of different fractions obtained after separation, and the antioxidant activity of each fraction was evaluated. The crude Asenticosus polysaccharides(ASPS)were obtained from the tender stems and leaves of ‘Zijia 1' by water extraction and alcohol precipitation, which were then separated and purified by DEAE-Cellulose 52 ion column and Sephadex G-100 gel column to obtain a uniform component. The ion chromatography and gel permeation chromatography-refractive index-multi-angle laser light scattering method was exploited to determine the monosaccharide compositions and molecular weight of the polysaccharides fractions. The hydroxyl radical(·OH), superoxide radical(O₂^-·)and 1,1-diphenyl-2-picrylhydrazyl radical(DPPH)scavenging ability were determined to evaluate the antioxidant of each fraction in vitro. Four polysaccharides ASPA-1-1, ASPA-2-1, ASPA-3-1 and ASPN-1 were isolated and purified from ASPS, with molecular weight of 8.10, 26.15, 0.91, 0.89 kDa, respectively, mainly composed of arabinose, rhamnose, galactose, glucose, xylose, mannose, ribose, galacturonic acid and glucuronic acid in different proportions. The ASPA-1-1, ASPA-2-1, ASPA-3-1 and ASPN-1 from ‘Zijia 1' demonstrated significant scavenging activities on ·OH, O₂^-· and DPPH free radical, the ability of ASPA-2-1 to scavenge ·OH and DPPH is higher than ASPA-1-1, ASPA-3-1 and ASPN-1; ASPA-3-1 has the strongest ability to scavenge O₂^-·. Therefore, the purified polysaccharides from ‘Zijia 1' has obvious antioxidant activity, which provides a scientific theoretical basis for its further utilization.