Optimization of Preparation Technology of Lactiplantibacillus plantarum HCS03-001 Freeze-dried Powder and its Anti-Helicobacter pylori Function

doi:10.19586/j.2095-2341.2023.0125

Current Biotechnology ›› 2024, Vol. 14 ›› Issue (2): 287-294.DOI: 10.19586/j.2095-2341.2023.0125

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Optimization of Preparation Technology of Lactiplantibacillus plantarum HCS03-001 Freeze-dried Powder and its Anti-Helicobacter pylori Function

Haitao WANG¹^,²(), Jia SONG¹^,², Ping YU¹^,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

Received:2023-10-17 Accepted:2024-02-04 Online:2024-03-25 Published:2024-04-17
Contact: Ping YU

植物乳植杆菌HCS03-001冻干粉制备工艺优化及抗幽门螺杆菌的功效

汪海涛¹^,²(), 宋佳¹^,², 余萍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

通讯作者: 余萍
作者简介:汪海涛 E-mail: 724312941@qq.com；

Abstract

Abstract:

The aim of the study is to optimize the preparation technology of freeze-dried powder of Lactiplantibacillus plantarum HCS03-001 and explore its functional effect of anti-Helicobacter pylori. In this study， the preparation technology of freeze-dried powder of Lactiplantibacillus plantarum HCS03-001 was optimized by single factor test， principal factor analysis and response surface test， and then the anti-Helicobacter pylori effect of the probiotics was verified by bacteriostatic test in vitro. The results showed that the optimum conditions for preparing freeze-dried powder were fermentation temperature 37 ℃， fermentation time 16 h， inoculum 5.2% and freeze-drying time 44 h. Under these conditions， the number of live bacteria was 5.85 × 10¹¹ CFU·g^-1. These results of bacteriostatic test in vitro showed that the fermentation supernatant and bacterial suspension of Lactiplantibacillus plantarum HCS03-001 could effectively inhibit the growth of Helicobacter pylori， and the inhibition rate was 69.55%~78.78%， which showed that Lactiplantibacillus plantarum HCS03-001 have the function of anti-Helicobacter pylori.

Key words: Lactiplantibacillus plantarum, freeze-dried powder, response surface, Helicobacter pylori

摘要：

为优化植物乳植杆菌（Lactiplantibacillus plantarum）HCS03-001的冻干粉制备工艺，探究其抗幽门螺杆菌的功能效果，通过单因素试验、主因素分析和响应面试验优化植物乳植杆菌HCS03-001冻干粉的制备工艺，采用体外抑菌试验验证该益生菌的抗幽门螺杆菌功效。结果发现，优化后制备冻干粉的最佳工艺条件为：发酵温度37 ℃，发酵时间16 h，接种量5.2%，冻干时间44 h，在此条件下活菌数达到5.85×10¹¹ CFU·g^-1。体外抑菌试验结果表明植物乳植杆菌HCS03-001的发酵上清液和菌悬液均能有效抑制幽门螺杆菌的生长，抑制率达到69.55%~78.78%。研究结果表明植物乳植杆菌HCS03-001具备抗幽门螺杆菌的功能。

关键词: 植物乳植杆菌, 冻干粉, 响应面, 幽门螺杆菌

CLC Number:

Q815

Haitao WANG, Jia SONG, Ping YU, Xuejiao CHEN. Optimization of Preparation Technology of Lactiplantibacillus plantarum HCS03-001 Freeze-dried Powder and its Anti-Helicobacter pylori Function[J]. Current Biotechnology, 2024, 14(2): 287-294.

汪海涛, 宋佳, 余萍, 陈雪娇. 植物乳植杆菌HCS03-001冻干粉制备工艺优化及抗幽门螺杆菌的功效[J]. 生物技术进展, 2024, 14(2): 287-294.

Figures/Tables 10

References 20

1	SAVOLDI A, CARRARA E, GRAHAM D Y, et al.. Prevalence of antibiotic resistance in Helicobacter pylori: a systematic review and meta-analysis in World Health Organization regions[J]. Gastroenterology, 2018, 155(5): 1372-1382.
2	LOSURDO G, CUBISINO R, BARONE M, et al.. Probiotic monotherapy and Helicobacter pylori eradication: a systematic review with pooled-data analysis[J]. World J. Gastroenterol., 2018, 24(1): 139-149.
3	刘昆梅,刘宏鹏,郭乐.幽门螺杆菌ATCC 43504感染BALB/c小鼠动物胃炎模型的建立及分析[J].生物技术进展,2015,5(6):446-450.
	LIU K M, LIU H P, GUO L. Establishment and analysis of gastritic model of Helicobacter pylori ATCC 43504 infection in BALB/c mice[J]. Curr. Biotechnol., 2015, 5(6): 446-450.
4	陈霞,夏晨梅,戴再友,等.十二指肠溃疡中幽门螺杆菌定植与菌群结构的关系[J].生物技术进展,2019,9(5):536-542.
	CHEN X, XIA C M, DAI Z Y, et al.. The relationship between Helicobacter pylori colonization and duodenal flora structure in duodenal ulcer[J]. Curr. Biotechnol., 2019, 9(5): 536-542.
5	MESTRE A, SATHIYA N R, RIVAS D, et al. Role of probiotics in the management of Helicobacter pylori [J/OL]. Cureus, 2022, 14(6): e26463 [2023-09-21]. .
6	朱红梅,刘颖初,薛元,等.2012-2022年中国幽门螺杆菌流行病学Meta分析[J].预防医学情报杂志,2023,39(9):1124-1137.
	ZHU H M, LIU Y C, XUE Y, et al.. Meta-analysis of epidemiology of Helicobacter pylori infection in China from 2012 to 2022[J]. J. Prev. Med. Inf., 2023, 39(9): 1124-1137.
7	PENUMETCHA S S, AHLUWALIA S, IRFAN R, et al.. The efficacy of probiotics in the management of Helicobacter pylori: a systematic review[J/OL]. Cureus, 2021, 13(12): e20483[2023-09-22]. .
8	CHEY W D, LEONTIADIS G I, HOWDEN C W, et al.. ACG clinical guideline: treatment of Helicobacter pylori infection[J]. Am. J. Gastroenterol., 2017, 112(2): 212-239
9	JI J, YANG H. Using probiotics as supplementation for Helicobacter pylori antibiotic therapy[J/OL]. Int. J. Mol. Sci., 2020, 21(3): 1136[2023-09-21]. .
10	QURESHI N, LI P, GU Q. Probiotic therapy in Helicobacter pylori infection: a potential strategy against a serious pathogen?[J]. Appl. Microbiol. Biotechnol., 2019, 103(4): 1573-1588.
11	BATDORJ B, TRINETTA V, DALGALARRONDO M, et al.. Isolation, taxonomic identification and hydrogen peroxide production by Lactobacillus delbrueckii subsp. lactis T31, isolated from Mongolian yoghurt: inhibitory activity on food-borne pathogens fool-borne pathogens[J]. J. Appl. Microbiol., 2007, 103(3): 584-593.
12	BAI X, ZHU M, HE Y, et al.. The impacts of probiotics in eradication therapy of Helicobacter pylori [J/OL]. Arch. Microbiol., 2022, 204(12): 692[2023-09-22]. .
13	汪海涛,汤纯,闵祥博,等.植物乳杆菌HCS03-001对龋病防治作用研究[J].中国乳品工业,2023,51(8):20-24.
	WANG H T, TANG C, MIN X B, et al.. Prevention and treatment effect on dental caries of Lactobacillus plantarum HCS03-001[J]. China Dairy Ind., 2023, 51(8): 20-24.
14	宋佳,余萍,矫艳平,等.具有缓解口臭效果的植物乳杆菌HCS03-001对食品防腐剂的耐受性研究[J].中国食品添加剂,2022,33(11):113-117.
	SONG J, YU P, JIAO Y P, et al.. Tolerance to food preservatives of Lactobacillus plantarum HCS03-001 with halitosis relieving effect[J]. China Food Addit., 2022, 33(11): 113-117.
15	陈雪,矫艳平,余萍,等.植物乳杆菌HCS03-001对小鼠功能性便秘及肠道菌群的影响[J].中国酿造,2022,41(1):55-58.
	CHEN X, JIAO Y P, YU P, et al.. Effect of Lactobacillus plantarum HCS03-001 on functional constipation and intestinal flora in mice[J]. China Brew., 2022, 41(1): 55-58.
16	矫艳平,余萍,赵迪,等.植物乳杆菌HCS03-001安全性评价及其益生特性分析[J].食品工业科技,2022,43(5):165-171.
	JIAO Y P, YU P, ZHAO D, et al.. Safety evaluation and probiotic characteristics analysis of Lactobacillus plantarum HCS03-001[J]. Sci. Technol. Food Ind., 2022, 43(5): 165-171.
17	周宇朦.维生素K2菌株的选育及高效发酵技术的研究[D].沈阳:沈阳农业大学,2020.
18	张立华,王一樊,张旭阳,等.响应面分析法优化酶法提取红景天苷的工艺研究[J].生物技术进展,2023, 3(3):441-448.
	ZHANG L H, WANG Y F, ZHANG X Y, et al.. Optimization of enzymatic extraction process of salidroside by response surface analysis[J]. Curr. Biotechnol. 2023, 13(3): 441-448.
19	管爱星,黄玉存,黄干荣,等.益生菌的不同组合体外抗幽门螺杆菌作用的实验研究[J].右江民族医学院学报,2023,45(2):208-212+217.
	GUAN A X, HUANG Y C, HUANG G R, et al.. Experimental study on anti-helicobacter pylori effect of different combinations of probiotics in vitro[J]. J. Youjiang Med. Univ. Natl., 2023, 45(2): 208-212+217.
20	张美怡,翟齐啸,赵建新,等.鼠李糖乳杆菌JS-SZ-2-1抗幽门螺杆菌感染的临床效果评价[J].食品与发酵工业,2020,46(23):29-34.
	ZHANG M Y, ZHAI Q X, ZHAO J X, et al.. Clinical efficacy evaluation of Lactobacillus rhamnosus JS-SZ-2-1 against Helicobacter pylori infection[J]. Food Ferment. Ind., 2020, 46(23): 29-34.

因素	水平
因素	-1	1
发酵温度/℃	35	36
发酵时间/h	8	12
初始pH	6.0	6.3
接种量/%	1	3
冻干保护剂添加比例	1∶1	1:2
冻干时间/h	40	42

因素	水平
因素	-1	1
发酵温度/℃	35	36
发酵时间/h	8	12
初始pH	6.0	6.3
接种量/%	1	3
冻干保护剂添加比例	1∶1	1:2
冻干时间/h	40	42

项目	平方和	自由度	均方	F值	P值
模型	6.73	14	0.48	1 344.61	<0.000 1
A-发酵温度	0.31	1	0.31	859.3	<0.000 1
B-发酵时间	0.15	1	0.15	412.33	<0.000 1
C-接种量	8.01×10^-3	1	8.01×10^-4	22.4	0.000 3
D-冻干时间	0.11	1	0.11	313.66	<0.000 1
AB	4.23×10^-3	1	4.23×10^-4	11.82	0.004
AC	1.23×10^-3	1	1.23×10^-4	3.43	0.085 4
AD	1.00×10^-4	1	1.00×10^-4	0.28	0.605 2
BC	0	1	0	0	1
BD	1.00×10^-4	1	1.00×10^-4	0.28	0.605 2
CD	0	1	0	0	1
A²	1.79	1	1.79	5 020.01	<0.000 1
B²	0.25	1	0.25	688.16	<0.000 1
C²	0.032	1	0.032	88.27	<0.000 1
D²	5.12	1	5.12	14 323.47	<0.000 1
残差	5.01×10^-3	14	3.58×10^-4	—	—
失拟项	3.13×10^-3	10	3.13×10^-4	0.66	0.726 9
纯误差	1.88×10^-3	4	4.70×10^-4	—	—
总离差	6.73	28	—	—	—

项目	平方和	自由度	均方	F值	P值
模型	6.73	14	0.48	1 344.61	<0.000 1
A-发酵温度	0.31	1	0.31	859.3	<0.000 1
B-发酵时间	0.15	1	0.15	412.33	<0.000 1
C-接种量	8.01×10^-3	1	8.01×10^-4	22.4	0.000 3
D-冻干时间	0.11	1	0.11	313.66	<0.000 1
AB	4.23×10^-3	1	4.23×10^-4	11.82	0.004
AC	1.23×10^-3	1	1.23×10^-4	3.43	0.085 4
AD	1.00×10^-4	1	1.00×10^-4	0.28	0.605 2
BC	0	1	0	0	1
BD	1.00×10^-4	1	1.00×10^-4	0.28	0.605 2
CD	0	1	0	0	1
A²	1.79	1	1.79	5 020.01	<0.000 1
B²	0.25	1	0.25	688.16	<0.000 1
C²	0.032	1	0.032	88.27	<0.000 1
D²	5.12	1	5.12	14 323.47	<0.000 1
残差	5.01×10^-3	14	3.58×10^-4	—	—
失拟项	3.13×10^-3	10	3.13×10^-4	0.66	0.726 9
纯误差	1.88×10^-3	4	4.70×10^-4	—	—
总离差	6.73	28	—	—	—

组别	OD₆₀₀	抑菌圈直径/mm	幽门螺杆菌抑制率/%
空白对照	—	0	0
阳性对照	—	20.92±1.02**	100.00**
菌悬液	1.911	16.48±2.14**	78.78**
发酵上清液	0.253	14.55±0.84**	69.55**

Optimization of Preparation Technology of Lactiplantibacillus plantarum HCS03-001 Freeze-dried Powder and its Anti-Helicobacter pylori Function

植物乳植杆菌HCS03-001冻干粉制备工艺优化及抗幽门螺杆菌的功效

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实验编号	发酵温度/℃	发酵时间/h	初始pH	接种量/%	冻干保护剂	冻干时间/h	活菌数/（×10¹¹ CFU·g^-1）
1	-1	-1	-1	-1	-1	-1	5.46
2	-1	1	-1	-1	-1	1	5.31
3	1	-1	1	1	-1	1	5.18
4	1	1	1	-1	1	1	5.11
5	1	1	-1	1	1	-1	5.06
6	-1	-1	-1	1	1	1	4.87
7	1	1	-1	1	-1	-1	5.25
8	1	-1	1	-1	-1	-1	5.29
9	-1	-1	1	1	1	-1	5.33
10	-1	1	1	-1	1	-1	5.02
11	1	-1	-1	-1	1	1	5.33
12	-1	1	1	1	-1	1	5.26

序号	A：发酵温度/℃	B：发酵时间/h	C：接种量/%	D：冻干时间/h	活菌数/（×10¹¹CFU·g^-1）
1	0	1	1	0	5.68
2	0	0	0	0	5.79
3	0	-1	1	0	5.48
4	0	-1	-1	0	5.44
5	0	0	0	0	5.81
6	0	0	-1	-1	4.74
7	0	0	-1	1	4.94
8	-1	1	0	0	5.09
9	0	-1	0	-1	4.52
10	1	-1	0	0	5.17
11	0	0	1	1	4.98
12	0	0	0	0	5.83
13	0	1	0	-1	4.75
14	1	0	0	-1	4.49
15	0	0	1	-1	4.78
16	0	1	-1	0	5.64
17	1	0	1	0	5.45
18	-1	0	1	0	5.08
19	1	1	0	0	5.33
20	1	0	-1	0	5.34
21	-1	0	0	1	4.34
22	0	0	0	0	5.84
23	0	-1	0	1	4.72
24	0	0	0	0	5.84
25	0	1	0	1	4.97
26	1	0	0	1	4.65
27	-1	0	0	-1	4.16
28	-1	-1	0	0	4.80
29	-1	0	-1	0	5.04

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