Preparation of Lactobacillus paracei TK1501 postbiotic and its inhibitory effect against Helicobacter pylori infection in mice

doi:10.12122/j.issn.1673-4254.2024.05.08

Abstract

Abstract:

Objective To prepare a postbiotic using soybean fermentation product of Lactobacillus paracasei TK1501 and evaluate its inhibitory effect against Helicobacter pylori (Hp) infection in mice. Methods L. paracasei TK1501 was cultured for 32 h at 37 ℃ in an anaerobic condition for solid substrate fermentation with a solid to water ratio of 1:1.5 in the substrate and an inoculation density of 5×10⁷ CFU/mL. The postbiotic was isolated and purified using macroporous resin XAD-16N adsorption, cation exchange chromatography and HPLC, and its stability and antibacterial activity were assessed. The inhibitory effect of this postbiotic against Hp infection was evaluated in a mouse model with gastric mucosal Hp infection, which were treated with the postbiotic via gavage for 4 weeks at the dose of 0.02 or 0.1 mL. Serum levels of TNF-α and IL-1β of the mice were analyzed after the treatments, and gastric tissues of the mice were collected for HE staining. Results L. paracasei TK1501 postbiotic could be easily degraded by protease and had good thermal stability and tolerance to exposures to acid, base, and organic solvents. In the in vitro experiment, the postbiotic showed strong inhibitory effects in bacterial cultures of Staphylococcus aureus, Hp and other common pathogenic bacteria without obviously affecting the resident bacteria in the digestive tract. In the mouse models, treatment with the postbiotic at the dose of 0.1 mL significantly alleviated Hp infection and lowered the serum levels of TNF-α and IL-1β of the mice. Conclusion L. paracasei TK1501 postbiotic has strong inhibitory effects on Hp and Staphylococcus aureus but not on normal intestinal flora in mice.

Key words: L. paracasei TK1501, isolation, Helicobacter pylori infection

Jinrui NIE, Yahui WU, Xuemei HAN, Yaqi LI, Haikuan WANG, Huitu ZHANG. Preparation of Lactobacillus paracei TK1501 postbiotic and its inhibitory effect against Helicobacter pylori infection in mice[J]. Journal of Southern Medical University, 2024, 44(5): 867-875.

Figures/Tables 13

References 39

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Protease types	Protease concentration (U/mL)
Protease types	0	10	20	30	50
Pepsin	25.1±0.3	21.3±0.4	19.3±0.1	16.7±0.3	15.8±0.2
Trypsase	25.3±0.5	19.6±0.3	16.8±0.1	15.3±0.3	13.7±0.3
chymotrypsin	24.8±0.1	21.5±0.5	14.7±0.4	10.8±0.6	10.7±0.4
Proteinase K	25.2±0.4	20.7±0.2	12.8±0.5	10.7±0.4	8.3±0.2
Papain	25.7±0.2	21.3±0.4	12.6±0.2	11.6±0.2	9.6±0.1
Alkaline protease	25.4±0.4	19.7±0.3	11.6±0.4	9.8±0.1	8.2±0.3

Protease types	Protease concentration (U/mL)
Protease types	0	10	20	30	50
Pepsin	25.1±0.3	21.3±0.4	19.3±0.1	16.7±0.3	15.8±0.2
Trypsase	25.3±0.5	19.6±0.3	16.8±0.1	15.3±0.3	13.7±0.3
chymotrypsin	24.8±0.1	21.5±0.5	14.7±0.4	10.8±0.6	10.7±0.4
Proteinase K	25.2±0.4	20.7±0.2	12.8±0.5	10.7±0.4	8.3±0.2
Papain	25.7±0.2	21.3±0.4	12.6±0.2	11.6±0.2	9.6±0.1
Alkaline protease	25.4±0.4	19.7±0.3	11.6±0.4	9.8±0.1	8.2±0.3

Indicator bacteria	Average number of colonies in the test group (CFU/mL)	Average colony number in control group (CFU/mL)	Sterilizing rate (%)
*S. aureus*	<10	4.5×10⁴	>99.98*
Hp	2.2×10⁴	6.5×10⁴	66.10
*Salmonella*	2.4×10⁴	5.5×10⁴	56.40
*C. jejuni*	1.6×10⁴	2.9×10⁴	44.80**
*C. perfringen*	2.2×10⁴	5.8×10⁴	63.80**
B. subtilis	5.5×10⁴	5.7×10⁴	3.50
Bifidobacterium	4.4×10⁴	4.7×10⁴	6.40
L. Beijerinck	5.6×10⁴	6.5×10⁴	13.80
E. coli	2.8×10⁴	3.1×10⁴	9.60
Bacteroides	3.8×10⁴	4.3×10⁴	24.30
C. Prazmowski	4.6×10⁴	4.7×10⁴	2.00
Proteus	3.5×10⁴	4.1×10⁴	14.60
Staphylococcus	2.1×10⁴	4.9×10⁴	57.10*
Streptococcus	4.3×10⁴	4.8×10⁴	10.40

Indicator bacteria	Average number of colonies in the test group (CFU/mL)	Average colony number in control group (CFU/mL)	Sterilizing rate (%)
*S. aureus*	<10	4.5×10⁴	>99.98*
Hp	2.2×10⁴	6.5×10⁴	66.10
*Salmonella*	2.4×10⁴	5.5×10⁴	56.40
*C. jejuni*	1.6×10⁴	2.9×10⁴	44.80**
*C. perfringen*	2.2×10⁴	5.8×10⁴	63.80**
B. subtilis	5.5×10⁴	5.7×10⁴	3.50
Bifidobacterium	4.4×10⁴	4.7×10⁴	6.40
L. Beijerinck	5.6×10⁴	6.5×10⁴	13.80
E. coli	2.8×10⁴	3.1×10⁴	9.60
Bacteroides	3.8×10⁴	4.3×10⁴	24.30
C. Prazmowski	4.6×10⁴	4.7×10⁴	2.00
Proteus	3.5×10⁴	4.1×10⁴	14.60
Staphylococcus	2.1×10⁴	4.9×10⁴	57.10*
Streptococcus	4.3×10⁴	4.8×10⁴	10.40

Groups	Feeding time (day)
Groups	0	14	21	28
Blank group	47.07±2.02**	47.92±2.21**	50.09±2.09*	53.53±2.62*
Model group	39.76±1.71	42.12±2.26	46.23±1.75	49.11±1.56
Experiment group 1	41.05±1.97	43.99±2.35	46.95±2.62	49.28±2.71
Experiment group 2	42.80±2.19 ^##	45.68±2.40 ^##	48.16±3.30	49.09±2.79
Experiment group 3	42.85±1.80 ^##	45.14±2.50 ^##	47.29±3.45	48.08±2.65