Presentation Information
[P01-051]Exploring the antimicrobial potential of lipopeptide biosurfactant produced by newly isolated bacteria against AMR pathogens
○Utpreksha Thapliyal1, Prof. Sangeeta Negi1 (1. Motilal Nehru National Institute of Technology Allahabad (India))
Keywords:
Lipopeptide,Anti-microbial resistance,ESKAPE pathogen,Minimum inhibitory concentration
Purpose: The efficiency of antimicrobial agents has come under investigation due to the increasing global rise of Antimicrobial-resistant (AMR) bacteria. These pathogens exhibit antimicrobial resistance mechanisms that has reduced the number of effective treatment choices. The application of biosurfactants is one of the novel approaches intended to reduce the prevalence of antimicrobial -resistant bacteria.
Method: Newly isolated bacteria were used to produce lipopeptide A1 and B4 respectively. The qualitative analysis of these newly isolated lipopeptide biosurfactants was tested by performing tests like emulsification index (EI24), oil displacement assay (ODA), drop collapse assay (DCA) and surface tension (ST). Later, the antimicrobial activity of lipopeptide A1, and B4 was tested against E. coli (MTCC6748), and 2 representative ESKAPE pathogens i.e. Staphylococcus aureus (MTCC3160), and Pseudomonas aeruginosa (MTCC424). The impact of these lipopeptides on the microbes was tested using three assays i.e. Broth dilution assay, Fluorescence-based viability assay, and Viable plate count assay. Further, minimum inhibitory concentration of the newly isolated lipopeptide was determined. The microbes were treated with different concentrations of lipopeptide i.e. 0.323g/l, 0.647g/l, 1.29g/l, 2.5g/l and 5.18g/l for 12hrs, 24hrs and 36hrs.
Results: The A1 was found to have 55% emulsification index while B4 showed 70%. The oil displacement assay of A1 and B4 was found to be 2cm and 2.5cm. The drop of A1 lipopeptide collapsed in 55sec while that B4 collapsed in 38sec. Using breakaway method, surface tension was calculated and it was found that A1, and B4 reduced the surface tension of water from 70mN/m to 30 mN/m and 28 mN/m respectively. The impact of A1 was found to be maximum on P. aeruginosa with the rfu and OD of 4294 and 0.865. Similarly, the lipopeptide B4 was found to have maximum antimicrobial effect on E. coli. The rfu and OD for E. coli was found to be 6100 and 1.590 respectively. The minimum inhibitory concentration of A1 against P. aeruginosa was found to be 5.18g/l after 36hrs of incubation and MIC of B4 on E. coli was found to be 2.5g/l after 24hrs of incubation. Scanning electron microscopy was performed to visualize the effect of lipopeptide on bacterial surface.
Conclusion: The application of lipopeptide biosurfactant as an anti-microbial agent has been less explored. Therefore, novel lipopeptide biosurfactants i.e. A1 and B4 have been isolated for the very first time isolated from oil contaminated sites and their activity was checked against disease causing pathogens.
Method: Newly isolated bacteria were used to produce lipopeptide A1 and B4 respectively. The qualitative analysis of these newly isolated lipopeptide biosurfactants was tested by performing tests like emulsification index (EI24), oil displacement assay (ODA), drop collapse assay (DCA) and surface tension (ST). Later, the antimicrobial activity of lipopeptide A1, and B4 was tested against E. coli (MTCC6748), and 2 representative ESKAPE pathogens i.e. Staphylococcus aureus (MTCC3160), and Pseudomonas aeruginosa (MTCC424). The impact of these lipopeptides on the microbes was tested using three assays i.e. Broth dilution assay, Fluorescence-based viability assay, and Viable plate count assay. Further, minimum inhibitory concentration of the newly isolated lipopeptide was determined. The microbes were treated with different concentrations of lipopeptide i.e. 0.323g/l, 0.647g/l, 1.29g/l, 2.5g/l and 5.18g/l for 12hrs, 24hrs and 36hrs.
Results: The A1 was found to have 55% emulsification index while B4 showed 70%. The oil displacement assay of A1 and B4 was found to be 2cm and 2.5cm. The drop of A1 lipopeptide collapsed in 55sec while that B4 collapsed in 38sec. Using breakaway method, surface tension was calculated and it was found that A1, and B4 reduced the surface tension of water from 70mN/m to 30 mN/m and 28 mN/m respectively. The impact of A1 was found to be maximum on P. aeruginosa with the rfu and OD of 4294 and 0.865. Similarly, the lipopeptide B4 was found to have maximum antimicrobial effect on E. coli. The rfu and OD for E. coli was found to be 6100 and 1.590 respectively. The minimum inhibitory concentration of A1 against P. aeruginosa was found to be 5.18g/l after 36hrs of incubation and MIC of B4 on E. coli was found to be 2.5g/l after 24hrs of incubation. Scanning electron microscopy was performed to visualize the effect of lipopeptide on bacterial surface.
Conclusion: The application of lipopeptide biosurfactant as an anti-microbial agent has been less explored. Therefore, novel lipopeptide biosurfactants i.e. A1 and B4 have been isolated for the very first time isolated from oil contaminated sites and their activity was checked against disease causing pathogens.
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