Exoproteome Analysis of Antagonistic Interactions between the Probiotic Bacteria Limosilactobacillus reuteri LR1 and Lacticaseibacillus rhamnosus F and Multidrug Resistant Strain of Klebsiella pneumonia

Abstract

The expansion of multiple drug resistant (MDR) strains of Klebsiella pneumoniae presents an immense threat for public health. Annually, this microorganism causes thousands of lethal nosocomial infections worldwide. Currently, it has been shown that certain strains of lactic acid bacteria (LAB) can efficiently inhibit growth of K. pneumoniae and the formation of its biofilms; however, the active principle of such action remains unknown. In the current article, the growth inhibition of MDR K. pneumoniae by two LAB—Limosilactobacillus reuteri LR1 and Lacticaseibacillus rhamnosus F—is demonstrated, and the nature of this inhibition studied at the level of exoproteome. This article shows that the exoproteomes of studied LAB contains both classically and non-classically secreted proteins. While for L. reuteri LR1 the substantial portion of classically secreted proteins was presented by cell-wall-degrading enzymes, for L. rhamnosus F only one out of four classically secreted proteins was presented by cell-wall hydrolase. Non-classically secreted proteins of both LAB were primarily metabolic enzymes, for some of which a possible moonlighting functioning was proposed. These results contribute to knowledge regarding antagonistic interaction between LAB and pathogenic and opportunistic microorganisms and set new perspectives for the use of LAB to control the spread of these microorganisms.

Oral Administration of Lacticaseibacillus rhamnosus CRL1505 Modulates Lung Innate Immune Response against Klebsiella pneumoniae ST25

“…The total number of leukocytes and differential cell counts in BAL samples were determined as described previously [ 13 , 14 ]. A hemocytometer was used to determine the total number of BAL and blood leukocytes.…”

“…BAL samples were obtained as described above. Blood samples were obtained via cardiac puncture and collected in heparinized tubes [ 14 , 15 ]. Cytokines determinations were performed in serum and BAL samples using commercial ELISA kits: IFN-γ (Mouse IFN-gamma Quantikine ELISA Kit, sensitivity: 2 pg/mL), and IL-10 (Mouse IL-10 Quantikine ELISA Kit, sensitivity: 5.2 pg/mL) from R&D Systems (Minneapolis, MN, USA), TNF-α (Mouse TNF alpha ELISA Kit, sensitivity: 9.1 pg/mL), IL-27 (Mouse IL-27 p28/IL30 Quantikine ELISA Kit, sensitivity: 4.7 pg/mL), IL-17 (Mouse IL-17 Quantikine ELISA Kit, sensitivity 5 pg/mL), IL-6 (Mouse IL-6 Quantikine ELISA Kit, sensitivity: 1.8 pg/mL), IL-8 mouse homolog chemokine KC (Mouse CXCL1/KC DuoSet ELISA, sensitivity 2.3 pg/mL), IL-1β (Mouse IL-1 beta ELISA Kit (ab197742), sensitivity: 1 pg/mL) and MCP-1 (Mouse MCP1 ELISA Kit (ab208979), sensitivity: 0.487 pg/mL) from Abcam (Cambridge, UK).…”

“…strains diminished the pathogen colonization and biofilm formation [ 8 , 9 , 10 , 11 , 12 ]. The ability of lactobacilli to inhibit the colonization of K. pneumoniae was associated with organic acid production [ 10 , 11 ] and the secretion of proteins with antimicrobial activity [ 13 , 14 ]. In addition, few studies reported that the oral administration of probiotic microorganisms to mice significantly reduced the lung inflammatory damage induced by K. pneumoniae nasal challenge [ 15 , 16 ], demonstrating that immunomodulatory bacteria are an interesting alternative to enhance resistance to this pathogen.…”

The impact of Elaeagnus angustifolia root exudates on Parafrankia soli NRRL B-16219 exoproteome

“…Additionally, the exoproteome can contain proteins that bind to host or microbial receptors, which allows for the mediation of signalling 41 ; 42 . Quorum sensing molecules are also present in the exoproteome and allow bacteria to coordinate their behaviour based on population density 39 ; 43 ; 44 . Some proteins within the exoproteome can also modulate the host immune system, suppressing host defences 39 ; 45 or promoting the growth of host tissues 46 - 48 .…”

Proteomic Analysis of Microorganisms

“…Based on the biophysical and biochemical comparative analysis of EVs, the authors suggested remarkable surface remodeling throughout of the T. cruzi life cycle. Savinova et al [2] focused on the antagonistic potential of two probiotic lactic acid bacteria (LAB) against MDR Klebsiella pneumoniae under co-culture conditions and used proteomic tools as to screen for major role prayers. Proteomic analysis of secreted proteins of L. reuteri LR1 and L. rhamnosus F revealed many classically or non-classically secreted proteins and suggested cell walldegrading enzymes and cell wall hydrolases as key enzymes for antagonistic interaction.…”