一株地衣芽孢杆菌发酵培养基优化及其液体制剂保存的探究

doi:10.11924/j.issn.1000-6850.casb2024-0496

摘要/Abstract

摘要：

本研究聚焦于地衣芽孢杆菌液体培养基及其液体制剂保存方法，旨在为地衣芽孢杆菌及其液体制剂的生产应用与深入研究提供理论依据。以地衣芽孢杆菌为试材，采用正交试验对液体培养基进行优化，用发酵罐试验对优化后的最佳培养基进行验证。同时，向地衣芽孢杆菌液体制剂中添加不同比例的植物乳杆菌，以探寻其最佳保存方法。研究结果显示：（1）地衣芽孢杆菌的最适培养基配方及发酵条件为麸皮10 g/L，豆粕15 g/L，碳酸钙0.75 g/L，氯化钠10 g/L；设定初始pH 7.5，温度37℃，转速200 r/min，在此条件下进行发酵，有效活菌数可达2.68×10¹⁰ cfu/mL。使用2 t发酵罐培养约24 h后发酵结束，此时有效活菌数能达到3.8×10¹⁰ cfu/mL。（2）在室温条件下，添加2.00%、5.00%植物乳杆菌的试验组中，地衣芽孢杆菌30 d后的活菌率分别为84.2%、84.1%，90 d后的活菌率分别达到了54.3%、52.4%。综上所述，本研究获得了该株地衣芽孢杆菌的高密度液体发酵培养基，为地衣芽孢杆菌液体制剂保存的相关研究奠定了理论基础。

关键词: 地衣芽孢杆菌, 发酵培养基, 液体制剂, 有效活菌数, 芽孢率, 麸皮, 植物乳杆菌

Abstract:

This research aims to establish a theoretical foundation for the production, application, and further exploration of Bacillus licheniformis and its liquid formulations by studying the liquid culture medium of Bacillus licheniformis and the preservation methods of its liquid preparations. Using Bacillus licheniformis as the test material, the liquid culture medium was optimized by orthogonal test, and the optimal medium was verified by fermenter test; different proportions of Lactobacillus plantarum were added to its liquid preparation to determine the best preservation method. The results were as follows. (1) The optimal medium and fermentation conditions were as following: bran 10 g/L, soybean meal 15 g/L, calcium carbonate 0.75 g/L, sodium chloride 10 g/L. The initial pH was 7.5, the temperature was 37℃, and the viable count of effective bacteria in the fermentation broth attained 2.68×10¹⁰ cfu/mL. The fermentation in a 2 t fermenter was completed after approximately 24 hours of cultivation, achieving a viable count of 3.8×10¹⁰ cfu/mL. (2) At indoor temperature, in the experimental groups supplemented with 2.00% and 5.00% Lactobacillus plantarum, the survival rates of Bacillus licheniformis were 84.2% and 84.1%, respectively, after 30 days, and reached 54.3% and 52.4% after 90 days. This study obtained a high-density liquid fermentation medium for this strain of Bacillus licheniformis, and laid a theoretical foundation for the research on the preservation of Bacillus licheniformis liquid preparations.

Key words: Bacillus licheniformis, fermentation medium, liquid preparations, effective viable bacteria count, spore rate, bran, Lactobacillus plantarum

叶伟伟, 谭新凤, 侯文赫, 魏善强, 颜晓晓, 张龙. 一株地衣芽孢杆菌发酵培养基优化及其液体制剂保存的探究[J]. 中国农学通报, 2025, 41(22): 110-116.

YE Weiwei, TAN Xinfeng, HOU Wenhe, WEI Shanqiang, YAN Xiaoxiao, ZHANG Long. Bacillus licheniformis: Fermentation Medium Optimization and Preservation of Its Liquid Preparation[J]. Chinese Agricultural Science Bulletin, 2025, 41(22): 110-116.

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参考文献 25

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序列	A.麸皮/(g/L)	B.豆粕/(g/L)	C.碳酸钙/(g/L)	菌体浓度/(×10⁹ cfu/mL)
1	5	5	0.25	9
2	5	10	0.5	5.5
3	5	15	0.75	19.5
4	10	5	0.5	16
5	10	10	0.75	22.8
6	10	15	0.25	25.6
7	15	5	0.75	16.5
8	15	10	0.25	13
9	15	15	0.5	15.5
I_j	34	41.5	56.5
II_j	62.4	48	37
III_j	45	60.6	58.8
K_j	3	3	3
I_j/k_j	11.3	13.8	18.8
II_j/k_j	20.8	16	12.3
III_j/k_j	15	20.2	19.6
R_j	9.5	6.4	7.3

序列	A.麸皮/(g/L)	B.豆粕/(g/L)	C.碳酸钙/(g/L)	菌体浓度/(×10⁹ cfu/mL)
1	5	5	0.25	9
2	5	10	0.5	5.5
3	5	15	0.75	19.5
4	10	5	0.5	16
5	10	10	0.75	22.8
6	10	15	0.25	25.6
7	15	5	0.75	16.5
8	15	10	0.25	13
9	15	15	0.5	15.5
I_j	34	41.5	56.5
II_j	62.4	48	37
III_j	45	60.6	58.8
K_j	3	3	3
I_j/k_j	11.3	13.8	18.8
II_j/k_j	20.8	16	12.3
III_j/k_j	15	20.2	19.6
R_j	9.5	6.4	7.3

时间/d	CK		乳酸菌添加量0.50%			乳酸菌添加量1.00%			乳酸菌添加量2.00%			乳酸菌添加量5.00%
时间/d	pH	a	pH	a	b	pH	a	b	pH	a	b	pH	a	b
7	8.5	52	8.5	12	1	8	22	2.1	7	57	6	7	63	7
15	8.5	46	8.5	10	1	8	17	1.4	7	59	5.6	7	63	7
22	8.5	29	8	1	1	7	14	1	6.5	55	5.2	6.5	60	6.5
30	8	24	8	0.6	0.14	7	8	1	6.5	48	5	6.5	53	5.7
60	8	14	7.9	0.3	0.015	7.88	0.8	0.7	6.56	39	3.8	6.29	41	4.5
90	7.2	8	7.48	0.3	0.01	7.29	0.4	0.04	6.29	31	1.9	6.06	33	2.4

时间/d	CK		乳酸菌添加量0.50%			乳酸菌添加量1.00%			乳酸菌添加量2.00%			乳酸菌添加量5.00%
时间/d	pH	a	pH	a	b	pH	a	b	pH	a	b	pH	a	b
7	8.5	52	8.5	12	1	8	22	2.1	7	57	6	7	63	7
15	8.5	46	8.5	10	1	8	17	1.4	7	59	5.6	7	63	7
22	8.5	29	8	1	1	7	14	1	6.5	55	5.2	6.5	60	6.5
30	8	24	8	0.6	0.14	7	8	1	6.5	48	5	6.5	53	5.7
60	8	14	7.9	0.3	0.015	7.88	0.8	0.7	6.56	39	3.8	6.29	41	4.5
90	7.2	8	7.48	0.3	0.01	7.29	0.4	0.04	6.29	31	1.9	6.06	33	2.4