徐丹鸿 王爽 林道鹏

摘要[目的]优化木瓜蛋白酶Sevag法去除酸浆（Physalis alkekengi L.）果实多糖中蛋白质的工艺条件。[方法]以蛋白脱除率和多糖保留率为指标，通过对酶解温度、时间、pH及酶添加量进行单因素试验及L9（34）正交试验确定酶解条件，并结合Sevag试剂除蛋白。[结果]各因素对酶解法脱酸浆果多糖液蛋白的影响程度大小依次为：酶解温度、酶解时间、pH、酶添加量。在酶解温度60 ℃、酶解时间2.0 h、pH 6.0、酶添加量2.5%条件下，粗多糖液蛋白脱除率为62.07%，多糖保留率为93.16%。原料经酶处理后，可使游离蛋白和部分与多糖结合的蛋白质被水解，再经Sevag试剂（氯仿∶正丁醇为4∶1）处理2次，蛋白质脱除率为81.65%，多糖保留率为89.43%，可达到较理想的脱蛋白质效果和较高的粗多糖保留率。[结论] 采用酶和Sevag试剂相结合的方法除酸浆果多糖中的蛋白质，效果良好，操作简单，可为酸浆果多糖的纯化提供理论依据，但其对多糖结构和生物活性的影响还有待于进一步研究。

关键词酸浆果多糖；脱蛋白；木瓜蛋白酶；Sevag法

中图分类号S509.9文献标识码A文章编号0517-6611（2016）04-113-03

Protein Removal from Polysaccharide by EnzymesSevag Process

XU Danhong， WANG Shuang， LIN Daopeng（Heilongjiang Ecological Engineering Professional College， Harbin， Heilongjiang 150025）

Abstract[Objective] The aim was to optimize conditions for removing protein from Physalis alkekengi polysaccharide with papainSevag method. [Method] With protein removal rate and polysaccharide retention rate as indicators， the single factor experiment was carried out by the enzyme hydrolysis temperature， time， pH and enzyme addition and L9 （34） orthogonal experiment. And combined Sevag reagent for removing protein. [Result] Influence of enzymatic deacidification of polysaccharide protein was the enzymolysis temperature > hydrolysis time>pH enzyme dosage. The optimum technological conditions were 60 ℃of hydrolysis temperature， hydrolysis time of 2.5 h， pH 6.0， enzyme adding amount of 2.5%， the removal rate of protein was 62.07% and the retention rate of polysaccharide was 93.16%. After the material processed by the enzyme， the free protein and the part of the protein which is combined with the polysaccharide can be hydrolyzed， and then treated for 2 times by Sevag reagent（chloroform Butyl alcohol 4∶1）， the removal rate of protein was 81.65% and the retention rate of polysaccharide was 89.43%. [Conclusion] Using papainSevag method to remove protein from Physalis alkekengi polysaccharide has a good effect and simple operation， which can provide theoretical basis for purification of Physalis alkekengi polysaccharide， but its effect on polysaccharide structure and biological activity remains to be further studied.

Key wordsPhysalis alkekengi L. polysaccharide； Removal of protein； Papain； Sevag method

酸漿（Physalis alkekengi L.）在我国分布广泛，尤其是在东北地区，具有对气候适应性强，耐寒、耐旱，易于人工栽培等特点，其全草皆可入药，是我国医书中记载的一种常用中药[1]。关于酸浆果实中的活性成分，已有学者对酸浆果实的多糖进行了分析和测定[2-3]，但从天然动植物中提取的多糖，常与蛋白形成糖蛋白复合物，使蛋白质的脱除比较困难，因此脱蛋白是多糖制备过程中一个重要的环节[4]。目前，国内对粗多糖脱蛋白的方法主要有Sevag法、三氟三氯乙烷法、外三氯乙酸法、酶法、盐酸法、树脂法等[5]。其中……