Study on Antibacterial Activity and Mechanism of Action of a Postbiotic Prepared by Co-fermenting Three Lactic Acid Bacteria against Methicillin-resistant <i>Staphylococcus aureus</i>

LAN Dongxue; QU Xinan; YU Haodong; HUANG Tian; PENG Chuantao; YAO Guoqiang; ZHAO Jinshan; LI Zhaojie

doi:10.13386/j.issn1002-0306.2023020029

Volume 44 Issue 21

Oct. 2023

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Science and Technology of Food Industry > 2023 > 44(21): 154-161. > DOI: 10.13386/j.issn1002-0306.2023020029

LAN Dongxue, QU Xinan, YU Haodong, et al. Study on Antibacterial Activity and Mechanism of Action of a Postbiotic Prepared by Co-fermenting Three Lactic Acid Bacteria against Methicillin-resistant Staphylococcus aureus[J]. Science and Technology of Food Industry, 2023, 44(21): 154−161. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023020029.

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Study on Antibacterial Activity and Mechanism of Action of a Postbiotic Prepared by Co-fermenting Three Lactic Acid Bacteria against Methicillin-resistant Staphylococcus aureus

LAN Dongxue^{1, 2, 3,},
QU Xinan^{1, 2, 3},
YU Haodong^{1, 2, 3},
HUANG Tian³,
PENG Chuantao^{1, 2},
YAO Guoqiang³,
ZHAO Jinshan^{1, 2},
LI Zhaojie^{1, 2, 3, ,}

1.
College of Food Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China
2.
Qingdao Special Food Research Institute, Qingdao 266109, China
3.
Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, Inner Mongolia Agricultural University, Hohhot 010018, China

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Received Date: February 05, 2023
Available Online: September 17, 2023

Graphical Abstract

Abstract

Abstract

To prepare a postbiotic with good antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA), and to study its antibacterial effect and action mode, a postbiotic named YDFF was prepared by co-fermenting Lactococcus lactis subsp Postbio-F3, Lactobacillus paracasei Postbio-P6 and Lactobacillus fermentans Postbio-Q7, its antibacterial activity against MRSA was determined by both inhibition zone and growth curve, and the active ingredients exerting antibacterial activity were preliminarily identified. The antibacterial mechanism of YDFF on MRSA was investigated by detecting the effects of YDFF on MRSA biofilm formation, ultrastructural changes, DNA leakage, and ROS generation. Furthermore, the effect of YDFF on the antibiotic resistance of MRSA to methicillin was explored by chessboard method. The results showed that the postbiotic YDFF had good antibacterial activity against MRSA (The diameter of inhibition zone was 19.1±1.5 mm) with wide pH tolerance range (pH3.0~9.0). The extracellular polysaccharides extracted from YDFF had no antibacterial activity, but protease K could completely inactivate its antibacterial activity, indicating that the main antibacterial active substance in YDFF was protein. It was demonstrated that YDFF exhibited antibacterial activity by several ways including inhibiting the formation of biofilms, destroying the cell membrane integrity which resulted in the leakage of DNA, and increasing the concentration of intracellular ROS. Additionally, when YDFF was used combined with methicillin, the fractional inhibitory concentration index (FICI) was significantly decreased to 0.38, indicating YDFF could reduce the antibiotic resistance of MRSA to methicillin. Taken together, YDFF exhibited good antibacterial activity on MRSA by inhibiting the biofilm formation, destroying membrane structure and increasing the intracellular ROS, and reduced the antibiotic resistance to methicillin, which would provide a theoretical basis for the use of postbiotics in food and medicine.
- postbiotics,
- methicillin-resistant Staphylococcus aureus,
- antibacterial,
- antibacterial mechanism,
- biofilm,
- antibiotic resistance

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References

[1]	SALMINEN S, COLLADO M C, ENDO A, et al. The International Scientific Association of Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of postbiotics[J]. Nature Reviews Gastroenterology & Hepatology,2021,18(9):649−667.
[2]	HOSSAIN M I, MIZAN M F R, ROY P K, et al. Listeria monocytogenes biofilm inhibition on food contact surfaces by application of postbiotics from Lactobacillus curvatus B. 67 and Lactobacillus plantarum M. 2[J]. Food Research International,2021,148:110595. doi: 10.1016/j.foodres.2021.110595
[3]	TOUSHIK S H, PARK J H, KIM K, et al. Antibiofilm efficacy of Leuconostoc mesenteroides J. 27-derived postbiotic and food-grade essential oils against Vibrio parahaemolyticus, Pseudomonas aeruginosa, and Escherichia coli alone and in combination, and their application as a green preservative in the seafood industry[J]. Food Research International,2022,156:111163. doi: 10.1016/j.foodres.2022.111163
[4]	DARWISH M S, QIU L, TAHER M A, et al. Health benefits of postbiotics produced by E. coli nissle 1917 in functional Yogurt enriched with cape gooseberry ( Physalis peruviana L)[J]. Fermentation,2022,8(3):128. doi: 10.3390/fermentation8030128
[5]	ŻOLLKIEWICZ J, MARZEC A, et al. Postbiotics—a step beyond pre-and probiotics[J]. Nutrients,2020,12(8):2189. doi: 10.3390/nu12082189
[6]	AGHEBATI-MALEKI L, HASANNEZHAD P, ABBASI A, et al. Antibacterial, antiviral, antioxidant, and anticancer activities of postbiotics:A review of mechanisms and therapeutic perspectives[J]. Biointerface Research in Applied Chemistry,2021,12:2629−2645. doi: 10.33263/BRIAC122.26292645
[7]	PUCCETTI M, XIROUDAKI S, RICCI M, et al. Postbiotic-enabled targeting of the host-microbiota-pathogen interface:Hints of antibiotic decline?[J]. Pharmaceutics,2020,12(7):624. doi: 10.3390/pharmaceutics12070624
[8]	ABD E G, WAFAA A. Paraprobiotics and postbiotics:Contemporary and promising natural antibiotics alternatives and their applications in the poultry field[J]. Open Veterinary Journal,2020,10(3):323−330.
[9]	李书鸿, 柳陈坚, 任贝贝, 等. 不同植物乳杆菌发酵液抑菌活性及其主要有机酸组成比较[J]. 食品科学,2019,40(5):8−16 LI S H, LIU C J, REN B B, et al. Comparison of bacteriostatic activity and main organic acid composition of fermentation broth of different Lactobacillus plantarum[J]. Food Science,2019,40(5):8−16.
[10]	XU C, FU Y, LIU F, et al. Purification and antimicrobial mechanism of a novel bacteriocin produced by Lactobacillus rhamnosus 1.0320[J]. LWT,2021,137:110338. doi: 10.1016/j.lwt.2020.110338
[11]	李杨, 周湘人, 郭薇丹, 等. 后生元的研究进展[J]. 食品安全质量检测学报,2021,12(16):6558−6564 LI Y, ZHOU X R, GUO W D, et al. Advances in the research of metagenesis[J]. Journal of Food Safety and Quality Inspection,2021,12(16):6558−6564.
[12]	CHUANTAO P, GUOQIANG Y, YARU S, et al. Comparative effects of the single and binary probiotics of Lacticaseibacillus casei Zhang and Bifidobacterium lactis V9 on the growth and metabolomic profiles in yogurts[J]. Food Research International,2022,152:110603. doi: 10.1016/j.foodres.2021.110603
[13]	JICHEMG W, HAOTIAN S, SHUAI G, et al. Comparison of the effects of single probiotic strains Lactobacillus casei Zhang and Bifidobacterium animalis ssp. lactis Probio-M8 and their combination on volatile and nonvolatile metabolomic profiles of yogurt[J]. Jounal of Dairy Science,2021,104(7):7509−7521. doi: 10.3168/jds.2020-20099
[14]	CHUNG H J, LEE H, KIM M, et al. Development and metabolic profiling of a postbiotic complex exhibiting antibacterial activity against skin microorganisms and anti-inflammatory effect on human keratinocytes[J]. Food Science and Biotechnology,2022,31(10):1325−1334. doi: 10.1007/s10068-022-01123-x
[15]	DU H, ZHOU L, LU Z, et al. Transcriptomic and proteomic profiling response of methicillin-resistant Staphylococcus aureus (MRSA) to a novel bacteriocin, plantaricin GZ1-27 and its inhibition of biofilm formation[J]. Applied Microbiology and Biotechnology,2020,104(18):7957−7970. doi: 10.1007/s00253-020-10589-w
[16]	GAJDACS M. The continuing threat of methicillin-resistant Staphylococcus aureus[J]. Antibiotics,2019,8(2):52. doi: 10.3390/antibiotics8020052
[17]	SHEHABELDINE A M, ASHOUR R M, OKBA M M, et al. Callistemon citrinus bioactive metabolites as new inhibitors of methicillin-resistant Staphylococcus aureus biofilm formation[J]. Journal of Ethnopharmacology,2020,254:112669. doi: 10.1016/j.jep.2020.112669
[18]	MORADI M, MOLAEI R, GUIMARAES J T. A review on preparation and chemical analysis of postbiotics from lactic acid bacteria[J]. Enzyme and Microbial Technology,2021,143:109722. doi: 10.1016/j.enzmictec.2020.109722
[19]	GOLKAR N, ASHOORI Y, HEIDARI R, et al. A novel effective formulation of bioactive compounds for wound healing:Preparation, in vivo characterization, and comparison of various postbiotics cold creams in a rat model[J]. Evidence-Based Complementary and Alternative Medicine,2021,2021:8577116−8577116.
[20]	MORADI, MEHRAN, KARIM M, et al. Characterization and application of postbiotics of Lactobacillus spp. on Listeria mo nocytogenes in vitro and in food models[J]. LWT,2019,111:457−464. doi: 10.1016/j.lwt.2019.05.072
[21]	史瑞军, 高静, 杜义强, 等. 抗菌肽Gal-4体外抑菌活性的初步鉴定[J]. 黑龙江畜牧兽医,2020(2):122−124 SHI R J, GAO J, DU Y Q, et al. Preliminary identification of antibacterial activity of antimicrobial peptide Gal-4 in vitro[J]. Heilongjiang Animal Husbandry and Veterinary,2020(2):122−124.
[22]	CAO H, MA S, GUO H, et al. Comparative study on the monosaccharide compositions, antioxidant and hypoglycemic activities in vitro of intracellular and extracellular polysaccharides of liquid fermented Coprinus comatus[J]. International Journal of Biological Macromolecules,2019,139:543−549. doi: 10.1016/j.ijbiomac.2019.08.017
[23]	杜宏, 吕欣然, 崔晓玲, 等. 融合魏斯氏菌对单增李斯特菌生物膜形成的影响[J]. 食品研究与开发,2021,42(13):180−187 DU H, LV X R, CUI X L, et al. Effect of Weisseria fusion on biofilm formation of Listeria monocytogenes[J]. Food Research and Development,2021,42(13):180−187.
[24]	刘萌, 魏莲花, 林赋桂, 等. ε-聚赖氨酸对耐甲氧西林金黄色葡萄球菌USA300的作用机制[J]. 中国抗生素杂志,2022,47(11):1−6 LIU M, WEI L H, LIN F G, et al. Mechanism of ε-polylysine on methicillin-resistant Staphylococcus aureus USA300[J]. Chinese Journal of Antibiotics,2022,47(11):1−6.
[25]	谷可欣, 张天翼, 何泾正, 等. 檀香醇对耐甲氧西林金黄色葡萄球菌USA300的抑制作用[J]. 湖南农业大学学报(自然科学版),2020,46(5):594−600 GU K X, ZHANG T Y, HE J Z, et al. Inhibitory effect of sandalwood on methicillin-resistant Staphylococcus aureus USA300[J]. Journal of Hunan Agricultural University (Natural Science Edition),2020,46(5):594−600.
[26]	YIN L, CHEN J, WANG K, et al. Study the antibacterial mechanism of cinnamaldehyde against drug-resistant Aeromonas hydrophila in vitro[J]. Microbial Pathogenesis,2020,145:104208. doi: 10.1016/j.micpath.2020.104208
[27]	LIU S, XU L, ZHANG T, et al. Oxidative stress and apoptosis induced by nanosized titanium dioxide in PC12 cells[J]. Toxicology,2010,267 (1-3):172−177. doi: 10.1016/j.tox.2009.11.012
[28]	WANG Y H, DONG H H, ZHAO F, et al. The synthesis and synergistic antifungal effects of chalcones against drug resistant Candida albicans[J]. Bioorganic & Medicinal Chemistry Letters,2016,26(13):3098−3102.
[29]	ROLTA R, SHARMA A, SOURIRAJAN A, et al. Combination between antibacterial and antifungal antibiotics with phytocompounds of Artemisia annua L:A strategy to control drug resistance pathogens[J]. Journal of Ethnopharmacology,2021,266:113420. doi: 10.1016/j.jep.2020.113420
[30]	HILAL S A, BUKET K. Antimicrobial activity of a bacteriocin produced by Enterococcus faecalis KT11 against some pathogens and antibiotic-resistant bacteria[J]. Food Science of Animal Resources,2018,38(5):1064−1079. doi: 10.5851/kosfa.2018.e40
[31]	李洁, 李晓然, 宫路路, 等. 乳酸片球菌发酵液中主要有机酸及其抑菌性研究[J]. 食品与发酵工业,2014,40(5):124−129 LI J, LI X R, GONG L L, et al. Study on the main organic acids in the fermentation broth of Lactococcus lactis and their antibacterial activity[J]. Food and Fermentation Industry,2014,40(5):124−129.
[32]	GEDEFIE A, DEMSIS W, ASHAGRIE M, et al. Acinetobacter baumannii biofilm formation and its role in disease pathogenesis:A review[J]. Infection and Drug Resistance,2021,10(14):3711−3719.
[33]	KUN C, CHUANTAO P, FANG C, et al. Antibacterial and antibiofilm activities of chlorogenic acid against Yersinia enterocolitica[J]. Frontiers in Microbiology,2022,13:1178.
[34]	WANG Z, ZHAI X, SUN Y, et al. Antibacterial activity of chlorogenic acid-loaded SiO₂ nanoparticles caused by accumulation of reactive oxygen species[J]. Nanology,2020,31(18):185101.
[35]	LONGFEI Y, LILI Z, ZHIMING M, et al. Antifungal effects of alantolactone on Candida albicans:An in vitro study[J]. Biomedicine & Pharmacotherapy,2022,149:112814.
[36]	PATHAK J, CHATTERJEE A, SINGH S P, et al. Detection of reactive oxygen species (ROS) in cyanobacteria using the oxidant-sensing probe 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA)[J]. Bio-protocol,2017,7 (17):e2545−e2545.