Purification of Polysaccharide Extract from Agaricus bisporus Using Macroporous Resins
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摘要: 对HZ830和D303两种大孔树脂串联纯化双孢蘑菇多糖的工艺进行了研究，考察体积流速、上样量和洗脱剂用量对双孢蘑菇多糖色素脱除率、蛋白脱除率和多糖保留率的影响。结果表明在以HZ830树脂为前柱，D303树脂为后柱，2种树脂等体积装柱，上样质量浓度为20 mg·mL-1（pH 8），上样体积5倍树脂柱床体积（5 BV，260 mL），体积流速为1.73 mL·min-1，上样结束后以1倍树脂柱床体积（1 BV，52 mL）纯水洗脱的工艺条件下，对蘑菇预煮液浓缩液中提取的双孢蘑菇粗多糖的蛋白质和色素杂质的脱除率分别为91.57%和94.14%，多糖保留率为73.49%。Abstract: Macroporous resins, HZ830 and D303, were used for the purification of polysaccharide extractfrom Agaricus bisporus. The evaluation criteria for the process included there movalrates of pigment and protein as well as the retention of polysaccharides in the filtrate under varied flow rates, feeding volumes and eluent flushing on the macroporous resin filtration system.The resin columns were assembled in series with either a HZ830 column in front or behind a D303 column. It was found that the best results could be achievedusing a HZ830 column followed by a D303 column andby feeding through themat a flow rate of 1.73 mL·min-1 of 5 times of the bed volume (BV, 260 mL) of a 20 mg·mL-1 polysaccharide extractat pH 8.0, the bed volume of colum were 52 mL, then flushing with 1 BV (52 mL) of water at the end. The resulting decolorization, deproteinization and polysaccharide retention rates of the filtration on thehot water mushroom extractreached 94.14%, 91.57% and 73.49%, respectively.
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Key words:
- Agaricus bisporus /
- macroporous resin /
- polysaccharides /
- decolorization /
- deproteinization
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图 1 HZ830静态吸附色素动力学曲线
注：不同小字字母表示差异达显著水平 (P＜0.05)。下图同。
Figure 1. Dynamic decolorization carves of types HZ830 macroporous resins
图 2 D303静态吸附色素动力学曲线
Figure 2. Dynamic decolorization carves of types D303 macroporous resins
图 3 体积流量对HZ830树脂脱色和脱蛋白效果的影响
Figure 3. Effect of filtration flow rate on pigment and protein removalsby HZ830 resin
图 4 体积流量对D303树脂脱色和脱蛋白效果的影响
Figure 4. Effect of filtrationflow rate on pigment and protein removals by D303 resin
图 5 HZ830树脂吸附容量与上样量的关系
Figure 5. Relationship between adsorption capacity of HZ830 and filtrationfeeding
图 6 D303树脂吸附容量与上样量的关系
Figure 6. Relationship between adsorption capacity of D303 and filtrationfeeding
图 7 洗脱剂用量对HZ830树脂中多糖洗脱效果的影响
Figure 7. Effect of eluent flushing on polysaccharide recovery by HZ830 resinfiltration
图 8 洗脱剂用量对D303树脂中多糖洗脱效果的影响
Figure 8. Effect of eluant volume on elution of polysaccharide on D303
表 1 HZ830和D303树脂柱串联处理前后脱色率、蛋白脱除率和多糖保留率测定结果
Table 1. Rates of decolorization, deproteinization and polysaccharides retention on infeed solutionbefore and after HZ830-D303 resin column treatment
项目 A460 A595 C蛋白/(μg·mL-1) A620 C糖/(μg·mL-1) 体积/mL 蛋白脱除率/% 色素脱除率/% 糖保留率/% 原液 1.721 1.801 43.4 0.787 99.15 260 - - - 中间取样 0.176 0.728 1.08 0.627 77.77 - 97.32 89.77 78.44 处理 0.13 0.736 2.12 0.534 65.38 290 94.14 91.57 73.49 注：考马斯法测定蛋白时样品稀释10倍；蒽酮法测定糖含量时样品稀释100倍；测定色素含量时不稀释。 
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