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从发酵食品中分离出的产类细菌素抑制物质的益生菌的广谱抗菌活性表征

Characterization of the broad-spectrum antibacterial activity of bacteriocin-like inhibitory substance-producing probiotics isolated from fermented foods.

作者信息

Thuy Tran Thi Dieu, Lu Hsu-Feng, Bregente Carl Jay Ballena, Huang Fong-Chi Annabelle, Tu Pei-Chun, Kao Cheng-Yen

机构信息

Institute of Microbiology and Immunology, College of Life Sciences, National Yang Ming Chiao Tung University, No. 155, Sec. 2, Linong Street, Taipei, 112, Taiwan.

Department of Laboratory Medicine, China Medical University Hospital, Taichung, Taiwan.

出版信息

BMC Microbiol. 2024 Mar 11;24(1):85. doi: 10.1186/s12866-024-03245-0.

Abstract

Antimicrobial peptides, such as bacteriocin, produced by probiotics have become a promising novel class of therapeutic agents for treating infectious diseases. Selected lactic acid bacteria (LAB) isolated from fermented foods with probiotic potential were evaluated for various tests, including exopolysaccharide production, antibiotic susceptibility, acid and bile tolerance, antibacterial activity, and cell adhesion and cytotoxicity to gastric cell lines. Six selected LAB strains maintained their high viability under gastrointestinal conditions, produced high exopolysaccharides, showed no or less cytotoxicity, and adhered successfully to gastric cells. Furthermore, three strains, Weissella confusa CYLB30, Lactiplantibacillus plantarum CYLB47, and Limosilactobacillus fermentum CYLB55, demonstrated a strong antibacterial effect against drug-resistant Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica serovar Choleraesuis, Enterococcus faecium, and Staphylococcus aureus. Whole genome sequencing was performed on these three strains using the Nanopore platform; then, the results showed that all three strains did not harbor genes related to toxins, superantigens, and acquired antimicrobial resistance, in their genome. The bacteriocin gene cluster was found in CYLB47 genome, but not in CYLB30 and CYLB55 genomes. In SDS-PAGE, the extract of CYLB30 and CYLB47 bacteriocin-like inhibitory substance (BLIS) yielded a single band with a size of less than 10 kDa. These BLIS inhibited the growth and biofilm formation of drug-resistant P. aeruginosa and methicillin-resistant S. aureus (MRSA), causing membrane disruption and inhibiting adhesion ability to human skin HaCaT cells. Moreover, CYLB30 and CYLB47 BLIS rescued the larvae after being infected with P. aeruginosa and MRSA infections. In conclusion, CYLB30 and CYLB47 BLIS may be potential alternative treatment for multidrug-resistant bacteria infections.

摘要

益生菌产生的抗菌肽,如细菌素,已成为治疗传染病的一类有前景的新型治疗剂。对从具有益生菌潜力的发酵食品中分离出的精选乳酸菌(LAB)进行了各种测试,包括胞外多糖的产生、抗生素敏感性、耐酸和耐胆汁性、抗菌活性以及对胃细胞系的细胞粘附和细胞毒性。六种精选的LAB菌株在胃肠道条件下保持高活力,产生高胞外多糖,显示无细胞毒性或细胞毒性较小,并成功粘附于胃细胞。此外,三种菌株,即孔氏魏斯氏菌CYLB30、植物乳杆菌CYLB47和发酵乳杆菌CYLB55,对耐药性大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌、猪霍乱沙门氏菌、粪肠球菌和金黄色葡萄球菌表现出强烈的抗菌作用。使用纳米孔平台对这三种菌株进行了全基因组测序;结果表明,这三种菌株的基因组中均未携带与毒素、超抗原和获得性抗菌耐药性相关的基因。在CYLB47基因组中发现了细菌素基因簇,但在CYLB30和CYLB55基因组中未发现。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)中,CYLB30和CYLB47细菌素样抑制物质(BLIS)提取物产生一条大小小于10 kDa的单一条带。这些BLIS抑制了耐药性铜绿假单胞菌和耐甲氧西林金黄色葡萄球菌(MRSA)的生长和生物膜形成,导致膜破坏并抑制其对人皮肤HaCaT细胞的粘附能力。此外,CYLB30和CYLB47 BLIS在感染铜绿假单胞菌和MRSA后拯救了幼虫。总之,CYLB30和CYLB47 BLIS可能是多药耐药细菌感染的潜在替代治疗方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dce4/10926564/4ab8ceae8854/12866_2024_3245_Fig1_HTML.jpg

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