响应面法优化刺五加高产异嗪皮啶的发酵工艺

doi:10.11924/j.issn.1000-6850.casb2021-0702

中国农学通报 ›› 2022, Vol. 38 ›› Issue (19): 152-158.doi: 10.11924/j.issn.1000-6850.casb2021-0702

所属专题：生物技术

• 食品·营养·检测·安全 • 上一篇下一篇

响应面法优化刺五加高产异嗪皮啶的发酵工艺

张赫¹^,²(), 万璐¹^,², 闫佳佳¹^,², 尤梦瑶¹^,², 曾伟民¹^,²(), 郑春英¹^,²()

¹黑龙江大学/农业微生物技术教育部工程研究中心,哈尔滨 150500
²黑龙江大学生命科学学院/黑龙江省普通高校微生物重点实验室,哈尔滨 150080

收稿日期:2021-07-21 修回日期:2021-10-14 出版日期:2022-07-05 发布日期:2022-07-13
通讯作者: 曾伟民,郑春英
作者简介:张赫,男,1999年出生,黑龙江哈尔滨人,硕士研究生,研究方向为食品和药物生物活性挖掘及研发。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学生命科学学院,Tel：0451-886608586,E-mail： 17082200652@163.com。
基金资助:
黑龙江大学校企合作横向项目“一株产原儿茶酸的枯草芽孢杆菌”(20170223)

Response Surface Method for Optimizing the Fermentation Process of Acanthopanax senticosus with High Isofraxidin Yield

ZHANG He¹^,²(), WAN Lu¹^,², YAN Jiajia¹^,², YOU Mengyao¹^,², ZENG Weimin¹^,²(), ZHENG Chunying¹^,²()

¹Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education/Heilongjiang University, Harbin 150500
²Key Laboratory of Microbiology, College of Heilongjiang Province/School of Life Sciences, Heilongjiang University, Harbin 150080

Received:2021-07-21 Revised:2021-10-14 Online:2022-07-05 Published:2022-07-13
Contact: ZENG Weimin,ZHENG Chunying

摘要/Abstract

摘要：

为提高发酵刺五加中异嗪皮啶的含量,采用单因素实验及响应面法对其发酵条件进行优化。采用单因素实验,分别测定发酵时间、初始pH、发酵温度、摇床转速、发酵菌株AJ14的菌悬液加入量及底物质量浓度对发酵刺五加中异嗪皮啶含量的影响,以获取单因素实验的最优值,然后采用响应面法中的Box-Behnken设计模块,分析初始pH、发酵温度、发酵菌株AJ14的菌悬液加入量3个自变量及其交互作用对异嗪皮啶得率的影响,确定最佳发酵工艺。结果表明,在初始pH 7.5、发酵温度27℃、发酵菌株AJ14菌悬液(1×10⁷CFU/mL)加入量6 mL的条件下,异嗪皮啶含量为(0.6629±0.0007) mg/g,与预测值的相对误差较小。研究结果可为采用微生物发酵法高效生产异嗪皮啶奠定基础,也可为其他中药发酵工艺提供数据参考。

关键词: 刺五加, 发酵, 异嗪皮啶, 单因素, 响应面法, 发酵工艺

Abstract:

Single factor experiment and response surface method were used to improve the yield of isofraxidin from fermenfed Acanthopanax senticosus. Fermentation duration, initial pH, fermentation temperature, shaker speed, added volume of AJ14 suspension and substrate concentration were detected to study the effects on the content of isofraxidin. Based on the single factor experiment, the optimized condition was determined by using Box-Behnken experimental design to study the initial pH, fermentation duration, added volume of AJ14 suspension and their interaction effects on the yields of isofraxidin. The results showed that the optimum parameters were initial pH of 7.5, fermentation temperature of 27℃ and the added volume of AJ14 suspension (1×10⁷CFU/mL) of 6 mL. Under the condition, isofraxidin in fermented A. senticosus was (0.6629±0.0007) mg/g and the relative error of predicted value was small. Therefore, the analytical results obtained by the response surface method could lay a foundation for the production of isofraxidin by microbial fermentation and provide reference for microbial fermented traditional Chinese medicine.

Key words: Acanthopanax senticosus, fermentation, isofraxidin, single factor, response surface method, fermentation process

中图分类号:

R931

张赫, 万璐, 闫佳佳, 尤梦瑶, 曾伟民, 郑春英. 响应面法优化刺五加高产异嗪皮啶的发酵工艺[J]. 中国农学通报, 2022, 38(19): 152-158.

ZHANG He, WAN Lu, YAN Jiajia, YOU Mengyao, ZENG Weimin, ZHENG Chunying. Response Surface Method for Optimizing the Fermentation Process of Acanthopanax senticosus with High Isofraxidin Yield[J]. Chinese Agricultural Science Bulletin, 2022, 38(19): 152-158.

图/表 12

参考文献 32

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序号	A.菌悬液加入量	B.pH	C.发酵温度	异嗪皮啶含量/(mg/g)
1	-1	1	0	0.460
2	1	1	0	0.440
3	0	0	0	0.520
4	0	0	0	0.540
5	-1	0	1	0.700
6	0	0	0	0.560
7	0	1	1	0.660
8	0	-1	-1	0.620
9	0	1	-1	0.500
10	1	0	-1	0.300
11	1	-1	0	0.560
12	1	0	1	0.640
13	0	-1	1	0.380
14	0	0	0	0.520
15	-1	-1	0	0.360
16	-1	0	-1	0.360
17	0	0	0	0.530

序号	A.菌悬液加入量	B.pH	C.发酵温度	异嗪皮啶含量/(mg/g)
1	-1	1	0	0.460
2	1	1	0	0.440
3	0	0	0	0.520
4	0	0	0	0.540
5	-1	0	1	0.700
6	0	0	0	0.560
7	0	1	1	0.660
8	0	-1	-1	0.620
9	0	1	-1	0.500
10	1	0	-1	0.300
11	1	-1	0	0.560
12	1	0	1	0.640
13	0	-1	1	0.380
14	0	0	0	0.520
15	-1	-1	0	0.360
16	-1	0	-1	0.360
17	0	0	0	0.530

来源	平方和	自由度	均方	F值	P	显著性
模型	0.27	9	0.029	15.52	<0.0001	^**
A	5.231×10^-3	1	5.231×10^-3	2.75	0.1280	^*
B	2.413×10^-8	1	2.413×10^-7	1.271×10^-4	0.9912
C	0.013	1	0.013	6.83	0.0259	^*
AB	1.800×10^-3	1	1.800×10^-3	0.95	0.3532
AC	0.051	1	0.051	26.96	0.0004	^**
BC	0.020	1	0.020	10.53	0.0088	^**
A²	0.065	1	0.065	33.97	0.0002	^**
B²	0.014	1	0.014	7.55	0.0205	^*
C²	0.12	1	0.12	63.75	<0.0001	^**
残差	0.019	10	1.899×10^-3
失拟项	0.015	5	3.051×10^-3	4.09	0.0743
纯误差	3.733×10^-3	5	7.467×10^-4
总和	0.28	19

来源	平方和	自由度	均方	F值	P	显著性
模型	0.27	9	0.029	15.52	<0.0001	^**
A	5.231×10^-3	1	5.231×10^-3	2.75	0.1280	^*
B	2.413×10^-8	1	2.413×10^-7	1.271×10^-4	0.9912
C	0.013	1	0.013	6.83	0.0259	^*
AB	1.800×10^-3	1	1.800×10^-3	0.95	0.3532
AC	0.051	1	0.051	26.96	0.0004	^**
BC	0.020	1	0.020	10.53	0.0088	^**
A²	0.065	1	0.065	33.97	0.0002	^**
B²	0.014	1	0.014	7.55	0.0205	^*
C²	0.12	1	0.12	63.75	<0.0001	^**
残差	0.019	10	1.899×10^-3
失拟项	0.015	5	3.051×10^-3	4.09	0.0743
纯误差	3.733×10^-3	5	7.467×10^-4
总和	0.28	19

响应面法优化刺五加高产异嗪皮啶的发酵工艺

Response Surface Method for Optimizing the Fermentation Process of Acanthopanax senticosus with High Isofraxidin Yield

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水平	因素
水平	A.菌悬液加入量/mL	B.pH	C.发酵温度/℃
-1	4	5	20
0	5	7	28
1	6	9	36

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