纤维素酶法提取决明子粗多糖的工艺优化及其抗氧化活性

蒋德旗; 许焯彬; 罗云; 李子强; 肖家宝; 莫烨; 李盈盈; 蒋丽林; 陈晓白

doi:10.13386/j.issn1002-0306.2022060262

纤维素酶法提取决明子粗多糖的工艺优化及其抗氧化活性

doi: 10.13386/j.issn1002-0306.2022060262

玉林师范学院生物与制药学院，广西高校亚热带生物资源保护与利用重点实验室，广西农产资源化学与生物技术重点实验室，广西玉林 537000

基金项目: 玉林市科学研究与技术开发计划项目（玉市科能20194302）；广西自然科学基金面上项目（2021GXNSFAA220001）；国家级大学生创新创业训练计划项目（202210606028）。

详细信息

作者简介:
蒋德旗（1986−），男，博士，教授，研究方向：天然产物活性成分开发与利用，E-mail：jiangdq8@sina.com

通讯作者:
蒋丽林（1986−），女，本科，助理研究员，研究方向：天然产物活性成分开发，E-mail：jianglilin55@126.com

中图分类号: R284.2
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出版历程
- 收稿日期: 2022-06-28
- 刊出日期: 2023-05-01

Optimization of Cellulase Extraction Process of Crude Polysaccharide from Semen Cassiae and Study on Its Antioxidant Activity

College of Biology and Pharmacy, Guangxi Education Department Key Laboratory for Conservation and Utilization of Subtropical Bio-resources, Guangxi Key Laboratory of Agricultural Resources Chemistry and Biotechnology, Yulin Normal University, Yulin 537000, China

摘要

摘要: 目的：优化纤维素酶法提取决明子粗多糖的工艺，并研究决明子粗多糖的体外抗氧化活性。方法：在单因素实验的基础上，以酶解时间、酶解温度、酶用量、液料比及酶解pH为自变量，多糖得率为响应值，利用Box-Behnken响应面法进行工艺优化。以对DPPH自由基和羟自由基清除率的大小为指标考察决明子粗多糖的体外抗氧化活性。结果：纤维素酶法提取决明子粗多糖最佳工艺为酶用量1.4%、酶解时间50 min、液料比24:1 mL/g、酶解pH5.4、酶解温度48 ℃，此条件下决明子多糖得率为11.67%，与回归模型的理论预测值11.91%误差小于5%。决明子粗多糖对DPPH自由基和羟自由基均具有较强的清除作用，半数抑制浓度分别为1.025 mg/mL和0.894 mg/mL。结论：纤维素酶法可显著提高决明子粗多糖得率，工艺简便可行，获得的决明子粗多糖具有体外抗氧化活性。
- 决明子 /
- 粗多糖 /
- 纤维素酶 /
- 抗氧化 /
- 响应面法
Abstract: Objective: To optimize the cellulase extraction process of crude polysaccharide from Semen Cassiae and study its antioxidant activity in vitro. Methods: On the basis of single factor test results, Box-Behnken response surface methodology was used to optimize the extraction process of crude polysaccharide from Semen Cassiae with enzymolysis time, enzymolysis temperature, enzyme dosage, liquid-to-material ratio and enzymolysis pH as independent variables, and the yield of polysaccharide as response value. DPPH radical and hydroxyl radical scavenging rates were used to investigate the antioxidant activities of crude polysaccharides from Semen Cassiae in vitro. Results: The optimal cellulase extraction conditions of crude polysaccharide from Semen Cassiae were as follows: Enzyme dosage was 1.4%, enzymolysis time was 50 min, liquid-to-material ratio was 24:1 mL/g, enzymolysis pH was 5.4, and enzymolysis temperature was 48 ℃. Under these conditions, the yield of polysaccharide from Semen Cassiae was 11.67%, and the theoretical prediction value of regression model was 11.91%, the error between the two conditions was less than 5%. Crude polysaccharide from Semen Cassiae had strong scavenging effects on DPPH free radical and hydroxyl free radical, with median inhibitory concentrations of 1.025 mg/mL and 0.894 mg/mL, respectively. Conclusion: Cellulase enzymatic method can significantly improve the yield of crude polysaccharide from Semen Cassiae, the process is simple and feasible, and the crude polysaccharide obtained from Semen Cassiae has antioxidant activity in vitro.
- Semen Cassia /
- crude polysaccharide /
- cellulose /
- antioxidant activity /
- response surface methodology

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图 1 酶用量对决明子多糖得率的影响

注：不同小写字母表示差异显著（P<0.05），图2~图5同。

Figure 1. Effects of enzyme addition on polysaccharide yield of Semen Cassia

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图 2 酶解温度对决明子多糖得率的影响

Figure 2. Effects of enzymolysis temperature on polysaccharide yield of Semen Cassia

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图 3 酶解pH对决明子多糖得率的影响

Figure 3. Effects of enzymolysis pH on polysaccharide yield of Semen Cassia

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图 4 酶解时间对决明子多糖得率的影响

Figure 4. Effects of enzymolysis time on polysaccharide yield of Semen Cassia

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图 5 液料比对决明子多糖得率的影响

Figure 5. Effects of liquid to material ratio on polysaccharide yield of Semen Cassia

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图 6 因素间交互作用的响应面图

Figure 6. Response surface diagram of interaction between factors

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图 7 决明子粗多糖对DPPH自由基的清除作用

Figure 7. Scavenging effects of crude polysaccharide from Semen Cassia on DPPH free radical

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图 8 决明子粗多糖对羟自由基的清除作用

Figure 8. Scavenging effects of crude polysaccharide from Semen Cassia on hydroxyl radical

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表 1 响应面设计因素与水平

Table 1. Experimental design factors and levels of response surface methodology

水平	因素
水平	A 酶解时间（min）	B 酶用量（%）	C 液料比（mL/g）	D 酶解pH
−1	50	0.8	16:1	4.6
0	80	1.2	20:1	5.0
1	110	1.6	24:1	5.4

下载: 导出CSV

表 2 响应面试验设计与结果

Table 2. Design and results of response surface test

试验号	A	B	C	D	多糖得率（%）
1	−1	0	0	1	11.2
2	0	1	0	−1	9.4
3	0	0	1	1	11.2
4	0	−1	1	0	10.1
5	0	0	0	0	10.3
6	−1	0	0	−1	9.2
7	−1	1	0	0	10.8
8	1	0	1	0	9.9
9	0	1	−1	0	10.2
10	0	−1	−1	0	9.9
11	0	1	1	0	10.9
12	0	0	0	0	10.7
13	0	−1	0	−1	8.8
14	0	1	0	1	10.7
15	0	0	0	0	11.1
16	0	0	−1	1	9.9
17	0	0	0	0	10.9
18	1	0	0	−1	9.7
19	1	−1	0	0	10.3
20	−1	0	−1	0	10.2
21	1	1	0	0	10.9
22	−1	−1	0	0	11.0
23	0	0	0	0	10.4
24	0	0	−1	−1	8.9
25	1	0	−1	0	9.9
26	1	0	0	1	10.0
27	0	−1	0	1	10.2
28	0	0	1	−1	9.3
29	−1	0	1	0	11.3

下载: 导出CSV

表 3 方差分析结果

Table 3. Results of ANOVA analysis

来源	平方和	自由度	均方	F值	P值	显著性
模型	12.62	14	0.90	11.76	<0.0001	**
A	0.83	1	0.83	10.79	0.0054	**
B	0.53	1	0.53	6.85	0.0203	*
C	1.22	1	1.22	15.87	0.0014	**
D	5.12	1	5.12	66.86	<0.0001	**
AB	0.16	1	0.16	2.09	0.1704
AC	0.32	1	0.32	4.24	0.0586
AD	0.71	1	0.71	9.32	0.0086	**
BC	0.06	1	0.06	0.75	0.4005
BD	0.01	1	0.01	0.09	0.7633
CD	0.14	1	0.14	1.88	0.1914
A²	0.01	1	0.01	0.19	0.6660
B²	0.08	1	0.08	1.06	0.3199
C²	0.59	1	0.59	7.66	0.0151	*
D²	3.03	1	3.03	39.54	<0.0001	**
残差	1.07	14	0.08
失拟项	0.62	10	0.06	0.55	0.8009
纯误差	0.45	4	0.11
总和	13.69	28
注：P<0.05显著，*P<0.01极显著。

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