Page 68 - 《精细化工》2023年第2期

第 40 卷第 2 期 精 细 化 工 Vol.40, No.2
20 23 年 2 月 FINE CHEMICALS Feb. 2023

中药现代化技术
碱提白芨粗多糖脱蛋白方法比较及抗氧化活性

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陈浩莹，王 斌 ，李金鹏，曾劲松，徐 峻，陈克复
（华南理工大学 轻工科学与工程学院，广东 广州 510641）

摘要：以碱提白芨粗多糖（BSP-A）为对象，对比三氯乙酸（TCA）法、TCA+正丁醇法、大孔树脂 AB-8 吸附
法、木瓜蛋白酶法、Sevage 法和三相分离法脱除 BSP-A 蛋白的效果。以 BSP-A 回收率和蛋白脱除率作为重要
指标，并探讨了蛋白脱除方法对 BSP-A 结构和生物活性的影响，确定更适用于 BSP-A 纯化的方法。结果表明，
Sevage 法、木瓜蛋白酶法和三相分离法对 BSP-A 的回收率达到 84%以上，反应相对温和；TCA 法、TCA+正丁
醇法、大孔树脂 AB-8 吸附法脱蛋白效果最佳，TCA+正丁醇法最高能达到 90.31%。不同的脱蛋白方法对 BSP-A
的重均相对分子质量影响轻微；而三相分离法和 TCA+正丁醇法影响了样品中 D-甘露糖和 D-葡萄糖的组成比例。
6 种方法均对 BSP-A 的 Zeta 电位和 FTIR 谱图的特征峰产生了一定的影响，其中，木瓜蛋白酶法和 Sevage 法能
较好地保留白芨多糖的抗氧化活性。总的来说，Sevage 法是一种对 BSP-A 结构和活性保留较好的脱蛋白方法，
但脱蛋白效率不佳，可作为白芨多糖脱除游离蛋白的有效手段之一。
关键词：白芨；粗多糖；脱蛋白方法；抗氧化；中药现代化技术
中图分类号：TQ281 文献标识码：A
文章编号：1003-5214 (2023) 02-0290-07 开放科学 (资源服务) 标识码 (OSID)：

Comparison of deproteinization methods of alkaline-extracted crude
polysaccharide from Bletilla striata and its antioxidant activity

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CHEN Haoying, WANG Bin , LI Jinpeng, ZENG Jinsong, XU Jun, CHEN Kefu
（School of Light Industry and Engineering, South China University of Technology, Guangzhou 510641, Guangdong, China）

Abstract: Trichloroacetic acid (TCA) method, TCA+n-butanol method, macroporous resin AB-8 adsorption
method, papain method, Sevage method, and three-phase separation method were compared for their
efficacy of protein removal from alkaline-extracted Bletilla Striata crude polysaccharide (BSP-A). The
effects of deproteinization methods on the structure and biological activity of BSP-A were discussed using
BSP-A recovery rate and protein removal efficacy as important indicators. Then, a more suitable method for
BSP-A purification was determined. The results showed that Sevage method, papain method, and
three-phase separation method all had a recovery rate for BSP-A more than 84%, with relatively mild
reaction conditions, while TCA method, TCA+n-butanol method, and macroporous resin AB-8 adsorption
method displayed better deproteinization results. TCA+n-butanol method could reached up to 90.31%.
Different deproteinization methods showed little effect on the weight-average relative molecular mass of
BSP-A, however, the three-phase separation method and TCA+n-butanol method affected the ratio of
D-mannose to D-glucose in BSP-A. These six methods demonstrated influence on Zeta potential and
characteristics peaks of FTIR spectrum of BSP-A. Especially, the papain and Sevage methods better
retained the antioxidant activity of polysaccharide from Bletilla striata. In general, Sevage method showed
good retention of polysaccharide structure and activity though with low deproteinization efficiency,
therefore, it could be one of the competing methods for free protein removal from BSP-A.

收稿日期：2022-07-18; 定用日期：2022-08-29; DOI: 10.13550/j.jxhg.20220675
基金项目：国家自然科学基金（22078113）；国家博士后创新人才支持计划（BX20200134）；广州市科技计划项目（202102080416，
202102020713）
作者简介：陈浩莹（1996—），女，博士生，E-mail：554505199@qq.com。联系人：王 斌（1985—），男，副研究员，博士，E-mail：
febwang@scut.edu.cn。