虾夷扇贝多糖提取及纯化方法的优化

Optimization of Extraction and Purification of Polysaccharides from Patinopecten yessoensis

以青岛虾夷扇贝为原料,利用单因素试验以及正交试验,对水提、酶提醇沉法结合超声波辅助提取扇贝柱多糖工艺及多糖脱蛋白进行研究,并对三氯乙酸(trichloroacetic acid,TCA)和D-葡萄糖酸-δ-内酯(D-glucono-δ-lactone,GDL)的脱蛋白效果进行对比,其中利用GDL对多糖中的蛋白质进行脱除尚属首次。结果显示,多糖提取最佳工艺条件为液料比40∶1(m L/g)、加酶量2.5%、60℃提取3 h,在此条件下虾夷扇贝多糖得率可达10.90%;TCA法脱蛋白的最佳工艺条件为脱蛋白时间5 h、TCA质量分数5%、脱蛋白次数3次;GDL法脱蛋白的最佳条件为反应时间1 h、反应温度45℃、GDL质量分数0.3%。在两者最佳条件下,分别进行3次平行实验,TCA蛋白脱除率为88.654%,相对标准偏差(relative standard deviation,RSD)为0.905%;GDL蛋白脱除率为99.062%,RSD为0.679%;对比两种脱蛋白方法,GDL法脱蛋白效率更高,实验效果也更为稳定。

In this paper,the ultrasonic-assisted enzymatic extraction and purification of polysaccharides from Patinopecten yessoensis were optimized using one-factor and orthogonal array design methods.For the deproteinization of crude polysaccharides,D-glucono-δ-lactone(GDL)was used in this study for the first time and its performance was evaluated in comparison with trichloroacetic acid(TCA).The results showed that the optimum conditions for extraction were as follows:solvent/solid ratio,40:1(mL/g);enzyme dosage,2.5%;and3h extraction at60℃,resulting in the highest polysaccharide yield of10.90%.The optimum conditions for TCA deproteinization were as follows:deproteinization time,5h;final TCA concentration,5%;and3deproteinization cycles.The optimal conditions for GDL deproteinization were as follows:deproteinization time,1h;reaction temperature,45℃;and final concentration,0.3%.Three parallel experiments conducted under the optimized conditions gave an average deproteinization rate of88.654%and99.062%with relative standard derivation(RSD)of0.905%and0.679%for TCA and GDL,respectively.Comparison of the two deproteinization methods showed that the deproteinization efficiency of GDL method was higher and more stable.