Optimization of Fermentation Culture Conditions for Cyclosporine A Using Response Surface Method

doi:10.19586/j.2095-2341.2025.0006

Current Biotechnology ›› 2025, Vol. 15 ›› Issue (3): 518-525.DOI: 10.19586/j.2095-2341.2025.0006

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Optimization of Fermentation Culture Conditions for Cyclosporine A Using Response Surface Method

Xiaojing GAO(), Zhulan ZHANG(), Xianju LIN, Guanrong QIU, Yaoming WEN, Jianhui FAN, Hongxiang HUANG

Fujian Provincial Key Laboratory of Screening for Novel Microbial Products，Fujian Institute of Microbiology，Fuzhou 350007，China

Received:2025-01-20 Accepted:2025-02-28 Online:2025-05-25 Published:2025-07-01
Contact: Zhulan ZHANG

基于响应面法优化环孢菌素A发酵工艺的研究

高晓晶(), 张祝兰(), 林仙菊, 邱观荣, 温耀明, 范建辉, 黄洪祥

福建省微生物研究所，福州 350007

通讯作者: 张祝兰
作者简介:高晓晶 E-mail: tju_gaoxiaojing@163.com；
基金资助:
福建省公益类科研院所专项(2023R1005002)

Abstract

Abstract:

Cyclosporin A （CsA） is a secondary metabolite with multiple biological activities such as immunosuppression， antibacterial， and anti-inflammatory effects， which holds significant application value in clinical treatment and the biomedical field. To further increase the fermentation content of CsA， based on single-factor experiments， the optimal fermentation conditions were obtained through Plackett-Burman design， central composite design （CCD） and response surface analysis （RSM）. The results indicated that the contents of corn steep powder， ammonium sulfate， glucose， and the fermentation time had a significant impact on the biosynthesis of CsA. Eventually， the suitable medium composition was determined as follows： 8.0% maltodextrin， 6.88% corn steep powder， 0.15% yeast extract powder， 0.18% ammonium sulfate， 0.61% glucose， and 0.1% PEG200. The optimized fermentation parameters were a cultivation temperature of 24 ℃ and a fermentation time of 10.8 days. The fermentation yield of CsA reached 11.76 mg·mL^-1， which was increased by 47.95% compared with the initial conditions， after the optimization of the fermentation process. The research results provide important theoretical basis and technical parameters for the process scale-up of this strain.

Key words: cyclosporin A, fermentation engineering, response surface methodology

摘要：

环孢菌素A（cyclosporin A，CsA）作为一种兼具免疫抑制、抗菌、抗炎等多种生物活性的次级代谢产物，在临床治疗和生物医药领域具有重要的应用价值。为了进一步提升CsA的发酵含量，以单因素试验为基础，采用Plackett-Burman设计、中心组合（central composite，CCD）设计及响应面分析法（response surface analysis，RSM）获取最优发酵条件。结果表明，玉米浆粉、硫酸铵、葡萄糖的含量及发酵时间对CsA的生物合成具有显著影响，最终确定适配的培养基组成为8.0%麦芽糊精、6.88%玉米浆粉、0.15%酵母浸粉、0.18%硫酸铵、0.61%葡萄糖和0.1% PEG200，最适的发酵温度和时间为24 ℃，和10.8 d。经发酵工艺优化后，CsA的发酵含量达11.76 mg·mL^-1，较初始条件提高了47.95%。研究结果可为该菌株的工艺放大提供重要的理论依据和技术参数。

关键词: 环孢菌素A, 发酵工程, 响应面法

CLC Number:

Q939.92

Xiaojing GAO, Zhulan ZHANG, Xianju LIN, Guanrong QIU, Yaoming WEN, Jianhui FAN, Hongxiang HUANG. Optimization of Fermentation Culture Conditions for Cyclosporine A Using Response Surface Method[J]. Current Biotechnology, 2025, 15(3): 518-525.

高晓晶, 张祝兰, 林仙菊, 邱观荣, 温耀明, 范建辉, 黄洪祥. 基于响应面法优化环孢菌素A发酵工艺的研究[J]. 生物技术进展, 2025, 15(3): 518-525.

Figures/Tables 11

References 23

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编码	组分	水平
编码	组分	-1	1
A	玉米浆粉/%	3	6
B	麦芽糊精/%	8	14
C	葡萄糖/%	0.5	1.5
D	酵母浸粉/%	0.1	0.2
E	PEG200/%	0.1	0.5
F	发酵时间/%	7	10
G	硫酸铵/%	0.1	1.0
H	发酵温度/℃	24	26

编码	组分	水平
编码	组分	-1	1
A	玉米浆粉/%	3	6
B	麦芽糊精/%	8	14
C	葡萄糖/%	0.5	1.5
D	酵母浸粉/%	0.1	0.2
E	PEG200/%	0.1	0.5
F	发酵时间/%	7	10
G	硫酸铵/%	0.1	1.0
H	发酵温度/℃	24	26

因素	编码	水平
因素	编码	-1	0	1
玉米浆粉/%	X₁	4	6	8
硫酸铵/%	X₂	0.1	0.2	0.3
发酵时间/d	X₃	9	10	11
葡萄糖/%	X₄	0.4	0.6	0.8

因素	编码	水平
因素	编码	-1	0	1
玉米浆粉/%	X₁	4	6	8
硫酸铵/%	X₂	0.1	0.2	0.3
发酵时间/d	X₃	9	10	11
葡萄糖/%	X₄	0.4	0.6	0.8

来源	平方和	自由度	均方	F值	P值
模型	37.39	8	4.67	9.87	0.043
A-玉米浆粉	9.32	1	9.32	19.68	0.021 3
B-麦芽糊精	4.36	1	4.36	9.22	0.056
C-葡萄糖	6.39	1	6.39	13.49	0.034 9
D-酵母浸粉	1.91	1	1.91	4.03	0.138 4
E-PEG200	0.045 4	1	0.045 4	0.095 8	0.777 2
F-发酵时间	8.18	1	8.18	17.27	0.025 3
G-硫酸铵	6.13	1	6.13	12.94	0.036 8
H-发酵温度	1.06	1	1.06	2.24	0.231 4
残差	1.42	3	0.473 5
总和	38.81	11

Optimization of Fermentation Culture Conditions for Cyclosporine A Using Response Surface Method

基于响应面法优化环孢菌素A发酵工艺的研究

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Figures/Tables 11

References 23

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编号	A	B	C	D	E	F	G	H	CsA含量/(mg·mL^-1)
1	1	-1	1	-1	1	1	-1	1	5.42
2	1	-1	-1	-1	1	-1	1	-1	6.69
3	-1	-1	-1	-1	-1	-1	-1	-1	6.53
4	1	1	1	-1	-1	-1	1	1	5.63
5	1	-1	1	1	1	-1	-1	1	7.45
6	-1	1	1	-1	1	1	1	-1	6.91
7	1	-1	1	1	-1	1	1	-1	7.65
8	-1	1	-1	1	1	-1	1	1	6.19
9	-1	-1	-1	1	-1	1	1	1	7.10
10	1	1	-1	1	1	1	-1	-1	11.44
11	1	1	-1	-1	-1	1	-1	1	10.89
12	-1	1	1	1	-1	-1	-1	-1	7.02

编号	玉米浆粉/%	葡萄糖/%	硫酸铵/%	发酵时间/d	CsA含量/(mg·mL^-1)
1	3	1.2	0.3	7	8.48
2	4	1.0	0.25	8	10.66
3	5	0.8	0.2	9	10.90
4	6	0.6	0.20	10	11.53
5	7	0.4	0.15	11	8.62
6	8	0.2	0.10	12	6.69
7	9	0	0.05	13	6.51

实验编号	X₁	X₂	X₃	X₄	CsA含量/(mg·mL^-1)
1	1	2	-1	1	8.94
2	0	0	0	0	11.92
3	-1	1	-1	1	8.84
4	1	-1	2	1	10.60
5	0	2	0	0	10.57
6	-1	-1	1	1	10.31
7	0	-2	0	0	10.12
8	-2	0	0	0	7.12
9	0	0	0	0	11.93
10	-1	1	1	1	9.59
11	2	0	0	0	7.87
12	1	-1	-1	-1	9.65
13	0	0	-2	0	8.75
14	1	1	1	-1	10.14
15	0	0	0	-2	10.04
16	0	0	0	0	11.77
17	0	0	0	0	11.30
18	-1	-1	-1	-1	9.71
19	-1	1	-1	-1	9.57
20	-1	-1	1	-1	10.50
21	1	-1	1	-1	10.92
22	0	0	2	0	11.23
23	1	1	1	1	10.90
24	-1	-1	-1	1	9.86
25	1	-1	-1	1	9.78
26	0	0	0	0	11.55
27	0	0	0	2	10.15
28	1	1	-1	-1	8.30
29	0	0	0	0	11.91
30	-1	1	1	-1	10.31

来源	平方和	自由度	均方	F值	P值
模型	39.52	14	2.82	14.37	<0.000 1	**
X₁-玉米浆粉	0.17	1	0.17	0.87	0.366 5
X₂-硫酸铵	0.62	1	0.62	3.14	0.096 7
X₃-发酵时间	7.68	1	7.68	39.08	<0.000 1	**
X₄-葡萄糖	0.000 1	1	0.000 1	0.000 6	0.981 3
X₁X₂	0.02	1	0.02	0.12	0.734 8
X₁X₃	0.63	1	0.63	3.18	0.094 7
X₁X₄	0.45	1	0.45	2.29	0.150 6
X₂X₃	0.24	1	0.24	1.22	0.286 4
X₂X₄	0.002 4	1	0.002	0.01	0.913 0
X₃X₄	0.03	1	0.028	0.14	0.709 1
X₁²	27.11	1	27.11	138.04	<0.000 1	**
X₂²	2.19	1	2.19	11.16	0.004 5	**
X₃²	3.77	1	3.77	19.20	0.000 5	**
X₄²	3.26	1	3.26	16.61	0.001 0	**
残差	2.95	15	0.20
失拟项	2.62	10	0.26	3.96	0.070 9
纯误差	0.33	5	0.066
总和	42.47	29

组别	CsA含量/（mg·mL^-1）
初始组	7.95±0.72
预测组	11.87±0.18*
验证组	11.76±0.47*

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