Study on Optimization of Culture Method and Improvement of Sleep Function of Limosilactobacillus reuteri HCS02-001

doi:10.19586/j.2095-2341.2023.0103

Current Biotechnology ›› 2024, Vol. 14 ›› Issue (1): 133-140.DOI: 10.19586/j.2095-2341.2023.0103

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Study on Optimization of Culture Method and Improvement of Sleep Function of Limosilactobacillus reuteri HCS02-001

Jia SONG¹^,²(), Xuelong LI³, Xinmei LIN¹^,², Ping YU¹^,², Sicheng YANG¹^,², Yongzhen PENG¹, Yanping JIAO¹^,²(), Xuejiao CHEN¹^,²

^1.Jiangxi Renren Health Microecological Technology Co. ，Ltd. ，Jiangxi Zhangshu 331200，China
^2.Renren Microbial Technology Research （Shenyang） Co. ，Ltd. ，Shenyang 110170，China
^3.Hangzhou Minsheng Health Pharmaceutical Co. ，Ltd. ，Hangzhou 310000，China

Received:2023-09-18 Accepted:2023-09-12 Online:2024-01-25 Published:2024-02-05
Contact: Yanping JIAO

罗伊氏粘液乳杆菌HCS02-001培养方法优化及改善睡眠功能研究

宋佳¹^,²(), 李雪龙³, 林欣梅¹^,², 余萍¹^,², 杨思程¹^,², 彭永振¹, 矫艳平¹^,²(), 陈雪娇¹^,²

^1.江西仁仁健康微生态科技有限公司，江西樟树 331200
^2.仁仁微生物科技研究（沈阳）有限公司，沈阳 110170
^3.杭州民生健康药业股份有限公司，杭州 310000

通讯作者: 矫艳平
作者简介:宋佳 E-mail: 573147127@qq.com；

Abstract

Abstract:

In order to optimize the cultivation method of Limosilactobacillus reuteri HCS02001 and explore its sleep promoting function， the optimal cultivation was screened through single factor experiment and response surface experiment based on the γ-aminobutyric acid （GABA） yield，and its sleep promoting function was evaluated through mouse experiment. Under the condition of that culture temperature was 37 ℃， the culture time was 24 h and the initial pH was 6.5， the GABA yield of L. reuteri HCS02-001 was 5.58 g·L^-1. No direct sleep phenomenon occurred in mice in the direct sleep experiment. The experimental results of prolonging the sleep time of mice hypnotized by pentobarbital sodium showed that the mice in the high-dose group effectively prolonged the sleep time， and the results of pentobarbital sodium subthreshold dose hypnosis experiment showed that the sleep rate of mice in the high-dose group was as high as 40%. Barbiturate sleep latency experiments showed that mice in the high-dose group had reduced sleep latency and could act synergistically with barbiturate sodium. The results showed that L. reuteri HCS02-001 had the function of sleep aid.

Key words: Limosilactobacillus reuteri, single factor, response surface, sleep aid

摘要：

为了优化罗伊氏粘液乳杆菌(Limosilactobacillus reuteri)HCS02-001的培养方法，并探究其助睡眠功能，以γ-氨基丁酸（γ-aminobutyric acid，GABA）的产量为考察指标，通过单因素实验、响应面实验和小鼠实验进行条件筛选和功能评价。在培养温度37 ℃，培养时间24 h，初始pH 6.5时，罗伊氏粘液乳杆菌的GABA产量为5.58 g·L^-1。直接睡眠实验中小鼠均未出现直接睡眠现象，延长戊巴比妥钠催眠小鼠的睡眠时间实验中，高剂量组的小鼠睡眠时间有效延长，戊巴比妥钠阈下剂量催眠实验结果显示高剂量组的小鼠入睡率高达40%，巴比妥那睡眠潜伏期实验显示高剂量组的小鼠减少了睡眠潜伏期，可与巴比妥钠协同发挥作用。结果表明罗伊氏粘液乳杆菌HCS02-001具备助睡眠的作用。

关键词: 罗伊氏粘液乳杆菌, 单因素, 响应面, 助睡眠

CLC Number:

Q935

Jia SONG, Xuelong LI, Xinmei LIN, Ping YU, Sicheng YANG, Yongzhen PENG, Yanping JIAO, Xuejiao CHEN. Study on Optimization of Culture Method and Improvement of Sleep Function of Limosilactobacillus reuteri HCS02-001[J]. Current Biotechnology, 2024, 14(1): 133-140.

宋佳, 李雪龙, 林欣梅, 余萍, 杨思程, 彭永振, 矫艳平, 陈雪娇. 罗伊氏粘液乳杆菌HCS02-001培养方法优化及改善睡眠功能研究[J]. 生物技术进展, 2024, 14(1): 133-140.

Figures/Tables 10

References 27

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项目	平方和	自由度	均方	F值	P
模型	10.77	9	1.20	219.95	<0.000 1
A	6.050×10^-3	1	6.050×10^-3	1.11	0.326 7
B	5.000×10^-5	1	5.000×10^-5	9.191×10^-3	0.926 3
C	0.061	1	0.061	11.26	0.012 2
AB	2.250×10^-4	1	2.250×10^-4	0.041	0.844 6
AC	3.025×10^-3	1	3.025×10^-3	0.56	0.480 1
BC	0.013	1	0.013	2.43	0.169 2
A²	6.74	1	6.74	1 238.07	<0.000 1
B²	2.27	1	2.27	417.85	<0.000 1
C²	0.78	1	0.78	142.95	<0.000 1
残差	0.038	7	5.440×10^-3
失拟项	0.026	3	8.600×10^-3	2.80	0.172 6
纯误差	0.012	4	3.070×10^-3
总离差	10.81	16

项目	平方和	自由度	均方	F值	P
模型	10.77	9	1.20	219.95	<0.000 1
A	6.050×10^-3	1	6.050×10^-3	1.11	0.326 7
B	5.000×10^-5	1	5.000×10^-5	9.191×10^-3	0.926 3
C	0.061	1	0.061	11.26	0.012 2
AB	2.250×10^-4	1	2.250×10^-4	0.041	0.844 6
AC	3.025×10^-3	1	3.025×10^-3	0.56	0.480 1
BC	0.013	1	0.013	2.43	0.169 2
A²	6.74	1	6.74	1 238.07	<0.000 1
B²	2.27	1	2.27	417.85	<0.000 1
C²	0.78	1	0.78	142.95	<0.000 1
残差	0.038	7	5.440×10^-3
失拟项	0.026	3	8.600×10^-3	2.80	0.172 6
纯误差	0.012	4	3.070×10^-3
总离差	10.81	16

分组	小鼠数量/只	睡眠数量/只	入睡率/%
空白对照组	10	0	0b
GABA水溶液组	10	1	10b
低剂量组	10	1	10b
中剂量组	10	2	20b
高剂量组	10	4	40a

分组	小鼠数量/只	睡眠数量/只	入睡率/%
空白对照组	10	0	0b
GABA水溶液组	10	1	10b
低剂量组	10	1	10b
中剂量组	10	2	20b
高剂量组	10	4	40a

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Study on Optimization of Culture Method and Improvement of Sleep Function of Limosilactobacillus reuteri HCS02-001

罗伊氏粘液乳杆菌HCS02-001培养方法优化及改善睡眠功能研究

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References 27

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因素	水平
因素	-1	0	1
A：培养温度/℃	36	37	38
B：培养时间/h	22	24	26
C：初始pH	6.3	6.5	6.7

试验号	A：培养温度	B：培养时间	C：初始pH	GABA产量/（g·L^-1）
1	0	0	0	5.66
2	-1	-1	0	3.56
3	0	-1	1	4.22
4	1	1	0	3.61
5	1	-1	0	3.65
6	0	0	0	5.62
7	1	0	-1	3.92
8	0	0	0	5.59
9	-1	0	1	3.82
10	0	0	0	5.58
11	-1	0	-1	3.94
12	1	0	1	3.91
13	0	1	1	4.35
14	0	0	0	5.51
15	0	1	-1	4.52
16	-1	1	0	3.55
17	0	-1	-1	4.62