生防贝莱斯芽孢杆菌SD23LJ1314发酵培养基的筛选及优化

doi:10.19586/j.2095-2341.2025.0114

生物技术进展 ›› 2026, Vol. 16 ›› Issue (1): 131-139.DOI: 10.19586/j.2095-2341.2025.0114

生防贝莱斯芽孢杆菌SD23LJ1314发酵培养基的筛选及优化

张梦华¹^,²(), 曲远航², 王培培²(), 董丽红², 张家齐², 付一帆², 杨威², 郭庆港², 李社增², 魏学军¹()

^1.河北工程大学园林与生态工程学院，河北邯郸 056038
^2.河北省农林科学院植物保护研究所，河北省农业有害生物综合防治技术创新中心，河北省作物有害生物综合防治国际科技联合研究中心，河北保定 071000

收稿日期:2025-08-27 接受日期:2025-10-24 出版日期:2026-01-25 发布日期:2026-02-12
通讯作者: 王培培,魏学军
作者简介:张梦华 E-mail: 2324224390@qq.com
基金资助:
国家重点研发计划项目(2022YFD1901300);河北省农林科学院现代农业科技创新专项(2022KJCXZX-ZBS-8)

Screening and Optimization of Fermentation Medium for Biocontrol Bacillus velezensis SD23LJ1314

Menghua ZHANG¹^,²(), Yuanhang QU², Peipei WANG²(), Lihong DONG², Jiaqi ZHANG², Yifan FU², Wei YANG², Qinggang GUO², Shezeng LI², Xuejun WEI¹()

^1.School of Landscape and Ecological Engineering，Hebei University of Engineering，Hebei Handan 056038，China
^2.Plant Protection Institute，Hebei Academy of Agriculture and Forestry Sciences，Integrated Pest Management Innovation Center of Hebei Province，International Science and Technology Joint Research Center on IPM of Hebei Province，Hebei Baoding 071000，China

Received:2025-08-27 Accepted:2025-10-24 Online:2026-01-25 Published:2026-02-12
Contact: Peipei WANG,Xuejun WEI

摘要/Abstract

摘要：

贝莱斯芽孢杆菌（Bacillus velezensis）SD23LJ1314是一株对辣椒疫病（Phytophthora capsici）具有很好防效的生防菌，其发酵水平的高低直接影响田间防治效果。为提升该菌株的发酵水平，以菌体量为核心评价指标，采用平板计数法对8种常用工业培养基的发酵菌量进行比较，筛选出了SD23LJ1314发酵的最适基础培养基。在此基础上，通过单因素实验对11种工业发酵常用碳源和9种氮源进行优化，确定最优碳氮源；进一步采用Plackett-Burman实验设计，并结合Box-Behnken设计与响应面分析法，对最优碳氮源的添加浓度进行了精确优化。结果表明，果糖与酵母粉分别为最优碳源和氮源组合，最终确定的发酵培养基配方为果糖10.75 g·L^-1、酵母粉21.48 g·L^-1、NaCl 5.65 g·L^-1、MnSO₄·7H₂O 0.71 g·L^-1。在此优化条件下，该菌株的发酵菌量达到9.32×10⁹ CFU·mL^-1，与基础培养基相比，菌体产量显著提高了31.6%。通过系列实验，明确了贝莱斯芽孢杆菌SD23LJ1314的高效发酵工艺，所获得的优化培养基配方及相关技术参数，为该生防菌的规模化生产奠定了坚实基础，对推动其应用于辣椒疫病的生物防治制剂开发具有重要价值。

关键词: 贝莱斯芽孢杆菌, 发酵液, 菌体量, 培养基优化, 响应面分析

Abstract:

Bacillus velezensis SD23LJ1314 is a biocontrol strain that has excellent efficacy against Phytophthoracapsici， and its fermentation level directly affects the field control effect. To enhance the fermentation level of this strain， this study used the bacterial quantity as the core evaluation index. Firstly， the fermentation bacterial quantity of 8 commonly used industrial culture media was compared using the plate counting method， and the most suitable basic culture medium for SD23LJ1314 fermentation was selected. On this basis， through single-factor experiments， 11 commonly used industrial fermentation carbon sources and 9 nitrogen sources were optimized to determine the optimal carbon and nitrogen sources. Subsequently， the Plackett-Burman experimental design was further adopted， combined with the Box-Behnken design and response surface analysis method， to precisely optimize the addition concentration of the optimal carbon and nitrogen sources. The experimental results showed that fructose and yeast powder were the optimal carbon source and nitrogen source combinations， respectively. After optimization by the above methods， the final optimized culture medium formula was determined as follows， 10.75 g·L^-1 fructose， 21.48 g·L^-1 yeast extract， 5.65 g·L^-1 NaCl， and 0.71 g·L^-1 MnSO₄·7H₂O. Under this optimized condition， the fermentation bacterial quantity of this strain reached 9.32×10⁹ CFU·mL^-1， and the bacterial yield was significantly increased by 31.6% compared to the basic culture medium. This study through a series of experiments clarified the efficient fermentation process of Bacillus velezensis SD23LJ1314. The optimized culture medium formula and related technical parameters obtained have laid a solid foundation for the large-scale production of this biocontrol strain and have significant value for promoting its application in the development of biological control agents for Phytophthora capsici.

Key words: Bacillus velezensis, fermentation broth, biomass, medium optimization, response surface analysis

中图分类号:

Q935

张梦华, 曲远航, 王培培, 董丽红, 张家齐, 付一帆, 杨威, 郭庆港, 李社增, 魏学军. 生防贝莱斯芽孢杆菌SD23LJ1314发酵培养基的筛选及优化[J]. 生物技术进展, 2026, 16(1): 131-139.

Menghua ZHANG, Yuanhang QU, Peipei WANG, Lihong DONG, Jiaqi ZHANG, Yifan FU, Wei YANG, Qinggang GUO, Shezeng LI, Xuejun WEI. Screening and Optimization of Fermentation Medium for Biocontrol Bacillus velezensis SD23LJ1314[J]. Current Biotechnology, 2026, 16(1): 131-139.

图/表 11

表1 芽孢杆菌基础培养基

Table 1 Basic mediums for Bacillus

培养基编号	配方
1	玉米粉30.0 g·L^-1、葡萄糖2.0 g·L^-1、黄豆粉20.0 g·L^-1、麸皮5.0 g·L^-1、 NaCl 4.0 g·L^-1
2	玉米粉30.0 g·L^-1、黄豆粉30.0 g·L^-1、K₂HPO₄·3H₂O 4.0 g·L^-1、MgSO₄·7H₂O 1.5 g·L^-1
3	葡萄糖10.0 g·L^-1、黄豆粉10.0 g·L^-1、NaCl 5.0 g·L^-1、MnSO4·7H₂O·0.6 g·L^-1
4	蛋白胨10.0 g·L^-1、酵母浸膏5.0 g·L^-1、葡萄糖1.0 g·L^-1、K₂HPO₄·3H₂O 0.2 g·L^-1、MgSO₄·7H₂O 0.2 g·L^-1
5	玉米粉50.0 g·L^-1、葡萄糖8.0 g·L^-1、豆粕粉20.0 g·L^-1、麸皮20.0 g·L^-1、NaCl 4.0 g·L^-1
6	玉米粉17.0 g·L^-1、葡萄糖10.0 g·L^-1、黄豆粉23.0 g·L^-1、麦芽糖10.0 g·L^-1、酵母浸膏5.0 g·L^-1
7	玉米粉5.0 g·L^-1、葡萄糖3.0 g·L^-1、豆饼粉5.0 g·L^-1、鱼粉3.0 g·L^-1
8	玉米粉40.0 g·L^-1、豆饼粉30.0 g·L^-1、酵母粉5.0 g·L^-1、K₂HPO₄·3H₂O 5.0 g·L^-1、MgSO₄·7H₂O 1.5 g·L^-1

表2 PB试验因素水平表

Table 2 Factors and levels of Plackett-Burman experiment

水平	因素/(g·L^-1)
水平	A：果糖	B：酵母粉	C	D：NaCl	E：MnSO₄·7H₂O	F
-1	10	10	-	5.0	0.6	-
1	15	15	-	7.5	0.9	-

表3 响应曲面试验因素水平表

Table 3 Factors and level design of Box-Behnken experiment

水平	因素/(g·L^-1)
水平	A：果糖	B：酵母粉	C：NaCl	D:MnSO₄·7H₂O
-1	10.0	15.00	5.00	0.60
0	12.5	18.75	6.25	0.75
1	15.0	22.50	7.50	0.90

图1 不同基础培养基对贝莱斯芽孢杆菌SD23LJ1314菌株菌体量的影响注：图中培养基序号所对应培养基同表1，不同小写字母表示在P<0.05水平上具有统计学意义。

Fig. 1 Effect of different basic mediums on the thallus concentration of B. velezensis strain SD23LJ1314

图2 不同碳源和氮源对贝莱斯芽孢杆菌SD23LJ1314菌体量的影响A:不同碳源对菌体量的影响；B:不同氮源对菌体量的影响。图中不同小写字母表示在P<0.05水平上具有统计学意义。

Fig. 2 Thallus concentrations of B. velezensis strain SD23LJ1314 in basic medium supplemented with different carbon and nitrogen sources

表4 Plackett-Burman试验设计与菌体量响应

Table 4 Experimental designs of Plackett-Burman and corresponding thallus concentrations

编号	A：果糖/(g·L^-1)	B：酵母粉/(g·L^-1)	D：NaCl/(g·L^-1)	E：MnSO₄·7H₂O/(g·L^-1)	菌体量/(10⁷ CFU·mL^-1)
1	15.0	15.0	7.5	0.6	178.00
2	10.0	15.0	7.5	0.6	712.40
3	10.0	10.0	5.0	0.9	445.20
4	15.0	15.0	5.0	0.9	133.60
5	15.0	10.0	7.5	0.9	44.50
6	15.0	10.0	7.5	0.6	89.10
7	15.0	15.0	5.0	0.9	160.30
8	10.0	15.0	7.5	0.9	801.40
9	10.0	15.0	5.0	0.6	756.90
10	30.0	10.0	5.0	0.6	356.20
11	15.0	10.0	5.0	0.6	106.90
12	10.0	10.0	7.5	0.9	400.70

表5 PB试验方差分析结果

Table 5 Analysis of variance of results of Plackett-Burman experiment

	自由度	平方和	均方	F	P
模型	4	785 619	196 405	23.06	0.001
线性	4	785 619	196 405	23.06	0.001
果糖(A)	1	635 013	635 013	74.55	0.001
酵母粉(B)	1	140 853	140 853	16.54	0.005
NaCl(D)	1	5 947	5 947	0.70	0.431
MnSO₄·7H₂O(E)	1	3 806	3 806	0.45	0.525
误差	2	59 629	8 518
合计	11	845 248
R²=0.929 5		R $a d j 2$ =0.889 1

表5 PB试验方差分析结果

Table 5 Analysis of variance of results of Plackett-Burman experiment

	自由度	平方和	均方	F	P
模型	4	785 619	196 405	23.06	0.001
线性	4	785 619	196 405	23.06	0.001
果糖(A)	1	635 013	635 013	74.55	0.001
酵母粉(B)	1	140 853	140 853	16.54	0.005
NaCl(D)	1	5 947	5 947	0.70	0.431
MnSO₄·7H₂O(E)	1	3 806	3 806	0.45	0.525
误差	2	59 629	8 518
合计	11	845 248
R²=0.929 5		R $a d j 2$ =0.889 1

表6 响应曲面试验设计及结果

Table 6 The design and results of response surface experiment

编号	果糖/(g·L^-1)	酵母粉/(g·L^-1)	NaCl/(g·L^-1)	MnSO₄·7H₂O/(g·L^-1)	响应值/(10⁷ CFU·mL^-1)
1	12.5	18.75	5.00	0.60	756.90
2	10.0	18.75	7.50	0.75	445.24
3	12.5	15.00	6.25	0.60	463.05
4	12.5	18.75	6.25	0.75	801.42
5	12.5	18.75	5.00	0.90	774.71
6	10.0	18.75	6.25	0.90	427.43
7	12.5	15.00	7.50	0.75	471.95
8	15.0	18.75	6.25	0.90	623.33
9	12.5	22.50	5.00	0.75	765.81
10	15.0	15.00	6.25	0.75	427.43
11	12.5	18.75	6.25	0.75	819.23
12	12.5	22.50	6.25	0.60	792.52
13	10.0	22.50	6.25	0.75	400.71
14	10.0	18.75	5.00	0.75	409.62
15	12.5	18.75	6.25	0.75	810.33
16	10.0	15.00	6.25	0.75	267.14
17	15.0	18.75	5.00	0.75	667.85
18	12.5	15.00	5.00	0.75	463.05
19	15.0	18.75	6.25	0.60	650.04
20	12.5	22.50	6.25	0.90	783.62
21	10.0	18.75	6.25	0.60	427.43
22	15.0	22.50	6.25	0.75	765.81
23	15.0	18.75	7.50	0.75	641.14
24	12.5	18.75	7.50	0.60	774.71
25	12.5	15.00	6.25	0.90	471.95
26	12.5	18.75	6.25	0.75	87.58
27	12.5	18.75	6.25	0.75	805.88
28	12.5	22.50	7.50	0.75	756.90
29	12.5	18.75	7.50	0.90	779.16

表7 Box-Behnken试验方差分析结果

Table 7 Analysis of variance of the results of Box-Behnken experiment

	自由度	平方和	均方	F	P
R²=0.982 9		R $a d j 2$ =0.970 8
模型	14	1 045 707.11	74 693.37	56.774 05	7.29E-10
A-果糖	1	205 408.33	205 408.33	156.129 8	5.56E-09
B-酵母粉	1	304 008.33	304 008.33	231.075 2	4.27E-10
C-NaCl	1	102.08	102.08	0.077 593	0.784 658
D-MnSO₄·7H₂O	1	2.08	2.08	0.001 584	0.968 82
AB	1	13 225.00	13 225.00	10.052 26	0.006 808
AC	1	1 225.00	1 225.00	0.931 116	0.350 946
AD	1	225	225	0.171 021	0.685 471
BC	1	100	100	0.076 01	0.786 808
BD	1	100	100	0.076 01	0.786 808
C	1	56.25	56.25	0.042 755	0.839 164
A²	1	425 236.77	425 236.77	323.220 3	4.54E-11
B²	1	168 223.25	168 223.25	127.865 7	2E-08
C²	1	9 325.45	9 325.45	7.088 228	0.018 575
D²	1	8 136.26	8 136.26	6.184 332	0.026 123
误差	14	18 418.75	1 315.63
纯误差	4	250	62.5
合计	28	2 199 749.66	1 211 445.30

表7 Box-Behnken试验方差分析结果

Table 7 Analysis of variance of the results of Box-Behnken experiment

	自由度	平方和	均方	F	P
R²=0.982 9		R $a d j 2$ =0.970 8
模型	14	1 045 707.11	74 693.37	56.774 05	7.29E-10
A-果糖	1	205 408.33	205 408.33	156.129 8	5.56E-09
B-酵母粉	1	304 008.33	304 008.33	231.075 2	4.27E-10
C-NaCl	1	102.08	102.08	0.077 593	0.784 658
D-MnSO₄·7H₂O	1	2.08	2.08	0.001 584	0.968 82
AB	1	13 225.00	13 225.00	10.052 26	0.006 808
AC	1	1 225.00	1 225.00	0.931 116	0.350 946
AD	1	225	225	0.171 021	0.685 471
BC	1	100	100	0.076 01	0.786 808
BD	1	100	100	0.076 01	0.786 808
C	1	56.25	56.25	0.042 755	0.839 164
A²	1	425 236.77	425 236.77	323.220 3	4.54E-11
B²	1	168 223.25	168 223.25	127.865 7	2E-08
C²	1	9 325.45	9 325.45	7.088 228	0.018 575
D²	1	8 136.26	8 136.26	6.184 332	0.026 123
误差	14	18 418.75	1 315.63
纯误差	4	250	62.5
合计	28	2 199 749.66	1 211 445.30

图3 果糖和酵母粉响应曲面图

Fig. 3 Fructose and yeast powder response surface chart

图4 贝莱斯芽孢杆菌SD23LJ1314菌株的生长曲线

Fig. 4 Growth curve of B. velezensis SD23LJ1314

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生防贝莱斯芽孢杆菌SD23LJ1314发酵培养基的筛选及优化

Screening and Optimization of Fermentation Medium for Biocontrol Bacillus velezensis SD23LJ1314

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