•  
  •  
 

Corresponding Author(s)

马夏吟(1991—),女,长沙理工大学副教授,博士。E-mail:[email protected]

Abstract

[Objective] To investigate the protective role of the surface protein LiaX in Lacticaseibacillus paracasei L9 against external stresses, such as bile salts, antibiotics, and the gastrointestinal environment. [Methods] A LiaX-overexpressing strain was constructed and the protein was purified. Spent media assays, in vitro simulated digestion, and exogenous protein supplementation experiments were comprehensively employed to evaluate the effect of LiaX on the survival rate of the strain under digestive, bile salt, and antibiotic stresses. [Results] The LiaX-overexpressing strain LPL9psip0968 his was successfully constructed, and the LiaX protein was isolated and purified. Spent media assays and exogenous addition of purified LiaX protein showed that the lysate of the LiaX-overexpressing strain exerted a significant protective effect on the bacteria under various stress conditions. In vitro simulated digestion experiments showed that LiaX significantly improved the survival rate of the strain during simulated gastric and intestinal digestion. [Conclusion] LiaX protein significantly enhances the stress resistance of the strain.

Publication Date

9-20-2026

First Page

132

Last Page

139

DOI

10.13652/j.spjx.1003.5788.2025.81085

References

[1] 宋静颐,张明,刘松玲,等.副干酪乳杆菌L9安全性评价 [J].中国奶牛,2015 (21):27-31.Song J Y,Zhang M,Liu S L,et al.Safety evaluation of Lactobacillus paracacei L9[J].China Dairy Cattle,2015 (21):27-31.

[2] 余萍,曹蓝,矫艳平,等.副干酪乳杆菌HCS17-040的筛选鉴定及其益生特性 [J].食品与机械,2021,37(11):28-33,129.Yu P,Cao L,Jiao Y P,et al.Screening of Lactobacillus paracasei HCS17-040 with triglyceride tunction and study on its probiotic characteristics [J].Food & Machinery,2021,37(11):28-33,129.

[3] 王然,李桂花,王鹏杰,等.灭活型副干酪乳杆菌L9常温酸奶调节肠道菌群缓解便秘作用研究 [J].食品科学技术学报,2020,38(3):95-101.Wang R,Li G H,Wang P J,et al.Effect of non-viable Lactobacillus paracasei L9 in normal temperature yogurt on modulating gut microbiota and relieving constipation [J].Journal of Food Science and Technology,2020,38(3):95-101.

[4] Sun N Y,Chen S B,Li Y X.Lactobacillus paracasei L9 ameliorated obesity-associated metabolic parameters and relevant gut microbiota in mice fed a high-fat diet [J].Nutrition Research,2023,115:26-37.

[5] 唐素婷,区锡敏,黄桂东,等.酱油渣中副干酪乳杆菌的分离鉴定及抗氧化特性研究 [J].食品与机械,2019,35(10):11-17,23.Tang S T,Ou X M,Huang G D,et al.Isolation,identification and antioxidant properties of Lactobacillus paracasei from soy sauce residue [J].Food & Machinery,2019,35(10):11-17,23.

[6] 胡敏,黄涛,彭珍,等.胆盐对植物乳杆菌NCU116应激基因和关键生理指标的影响 [J].食品与发酵工业,2019,45(9):1-8.Hu M,Huang T,Peng Z,et al.Influences of bile salts on stress genes and key physiological indexes of Lactobacillus plantarum NCU116[J].Food and Fermentation Industries,2019,45(9):1-8.

[7] 胡斌,田丰伟,赵建新,等.嗜酸乳杆菌NCFM表层蛋白的提取及其与菌体酸和胆盐耐受及粘附性能的关系 [J].食品工业科技,2014,35(5):118-121.Hu B,Tian F W,Zhao J X,et al.Extraction of surface layer protein from Lactobacillus acidophilus NCFM and the relationship among SLP and strain acid,bile resistance and cell adhesion [J].Science and Technology of Food Industry,2014,35(5):118-121.

[8] 王艳美,刘永乐,邓凯,等.LiaX家族表面蛋白LPL9_0968在副干酪乳杆菌L9胆盐胁迫应激中的作用 [J].中国食品学报,2022,22(12):104-115.Wang Y M,Liu Y L,Deng K,et al.Role of LiaX family surface protein LPL9_0968 in bile salt stress resistance in Lacticaseibacillus paracasei L9[J].Journal of Chinese Institute of Food Science and Technology,2022,22(12):104-115.

[9] Axell-House D B,Simar S R,Panesso D,et al.LiaX is a surrogate marker for cell envelope stress and daptomycin non-susceptibility in Enterococcus faecium [J].Antimicrobial Agents and Chemotherapy,2024,68(3):e0106923.

[10] Khan A,Davlieva M,Panesso D,et al.Antimicrobial sensing coupled with cell membrane remodeling mediates antibiotic resistance and virulence in Enterococcus faecalis [J].Proceedings of the National Academy of Sciences of the United States of America,2019,116(52):26 925-26 932.

[11] Ota Y,Furuhashi K,Hayashi W,et al.Daptomycin resistant Enterococcus faecalis has a mutation in LiaX,which encodes a surface protein that inhibits the LiaFSR systems and cell membrane remodeling [J].Journal of Infection and Chemotherapy,2021,27(1):90-93.

[12] Panesso Botero D,Singh K,Gomez-Villegas S I,et al.LiaF is required for LiaX mediated protection against Daptomycin and LL-37 in Enterococcus faecalis OG1RF[J].Open Forum Infectious Diseases,2022,9(Suppl 2):ofac492.157.

[13] Yan N N,Luo T,Wang Y M,et al.Transcriptomic and biochemical analyses reveal the cell membrane defense regulated by LiaFSR-LiaX system in Lacticaseibacillus paracasei L9 under bile salt stress [J].Food Bioscience,2024,62:105145.

[14] Brodkorb A,Egger L,Alminger M,et al.INFOGEST static in vitro simulation of gastrointestinal food digestion [J].Nature Protocols,2019,14(4):991-1 014.

[15] Liu H,Cui S W,Chen M S,et al.Protective approaches and mechanisms of microencapsulation to the survival of probiotic bacteria during processing,storage and gastrointestinal digestion:a review [J].Critical Reviews in Food Science and Nutrition,2019,59(17):2 863-2 878.

[16] Zhang Y,Xie Y F,Liu H,et al.Probiotic encapsulation in water-in-oil high internal phase emulsions:enhancement of viability under food and gastrointestinal conditions [J].LWT-Food Science and Technology,2022,163:113499.

[17] Kaur Sidhu M,Lyu F,Sharkie T P,et al.Probiotic yogurt fortified with chickpea flour:physico-chemical properties and probiotic survival during storage and simulated gastrointestinal transit [J].Foods,2020,9(9):1 144.

[18] 王露霏,王彩云,李嘉文,等.双歧杆菌四联活菌体外模拟消化液耐受性的考察 [J].中国微生态学杂志,2023,35(11):1 280-1 285,1 291.Wang L F,Wang C Y,Li J W,et al.The tolerance of Bifidobacterium-quadruple probiotics to the simulated digestive fluid in vitro [J].Chinese Journal of Microecology,2023,35(11):1 280-1 285,1 291.

[19] Vega L A,Sanson-Iglesias M,Mukherjee P,et al.LiaR-dependent gene expression contributes to antimicrobial responses in group A Streptococcus [J].Antimicrobial Agents and Chemotherapy,2024,68(12):e0049624.

[20] Tymoszewska A,Szylinska M,Aleksandrzak-Piekarczyk T.The LiaFSR-LiaX system mediates resistance of Enterococcus faecium to peptide antibiotics and to aureocin A53- and enterocin L50-like bacteriocins [J].Microbiology Spectrum,2023,11(3):e00343-23.

[21] Wu H,Xue E S,Zhi N,et al.d-Methionine and d-phenylalanine improve Lactococcus lactis F44 acid resistance and nisin yield by governing cell wall remodeling [J].Applied and Environmental Microbiology,2020,86(9):e02981-19.

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.