大孔吸附树脂对鹿托盘蛋白水解液脱盐及纯化的作用
2018-06-08姜薇
姜薇
【摘 要】为了脱除鹿托盘蛋白水解液中的盐分并分离出具有较高抗氧化活性的组分,本研究选择DA201-C大孔树脂对水解液中多肽的吸附能力进行测定。结果表明:当多肽浓度为10 mg/mL,上样流速1.0 BV/h,树脂对多肽的脱盐率达到98.64±2.45%,其中75%乙醇洗脱组分的抗氧化活性最高,此时鹿托盘多肽对DPPH·和ABTS·+的清除率分别为89.5±1.14%和86.3±1.47%
【关键词】鹿托盘;大孔吸附树脂;蛋白水解液;脱盐;分离纯化
中图分类号:TS251.92 文献标识码: A 文章编号: 2095-2457(2018)08-0045-002
Effect of Macroporous Adsorption Resin on Desalination and Purification of Deer Trays Protein Hydrolysate
JIANG Wei
(Huangshan College of Tourism, Huangshan, Anhui 245041, China)
【Abstract】In order to remove the salt from the hydrolysates of deer trays and isolate the components with higher antioxidant activity, the DA201-C macroporous resin was used to determine the adsorption capacity of polypeptides in the hydrolysate. The results showed that when the peptide concentration was 10 mg/mL and the sample flow rate was 1.0 BV/h, the desalination rate of the resin to the peptide reached 98.64±2.45%. Among them, the antioxidant activity of the 75% ethanol elution component was the highest. The clearance rates of DPPH· and ABTS·+ by peptides were 89.5±1.14% and 86.3±1.47%, respectively.
【Key words】Deer tray; Macroporous resin; Protein hydrolyzate; Desalting; Separation and purification
每年6月份为梅花鹿或马鹿的产鹿茸期,鹿茸被收割后会残留约3cm的根基,并于次年新鹿茸长出前脱落,因其形似一个圆盘,故名鹿托盘、鹿角冒[1]。鹿托盘中含有丰富的胶原蛋白,经碱性蛋白酶水解后可获得具有抗氧化活性的蛋白水解液,因在酶解的过程中需要不断的添加NaOH溶液来维持水解液pH的稳定性,从而导致水解中存在大量的盐分,盐分的存在会影响鹿托盘抗氧化肽的应用领域。
一般情况下,透析袋常被用来对大分子有机物进行透析除盐,但是实验过程要不断地更换缓冲溶液,具有耗时长、损失高、资源浪费等缺点。除此之外,还可以采用纳滤膜[2]、凝胶色谱法[3]对蛋白进行纯化,但它们都比较昂贵仅限于实验室研究。大孔树脂除盐具有低成本、高效并能保证产品活性的优点,已被广泛地用于生物样品的除盐[4]。大孔树脂对有机物的吸附作用,依靠于其与有机物疏水基团之间的范德华力,而无机盐不被大孔树脂吸附,采用去离子水就能将水解液中的盐分去除,通过添加合适的解析液就能改变吸附体系的疏水与亲水平衡,影响两者的吸附力大小,从而使具有不同疏水基团的有机物被解析下来[5]。……
