Mining of plant-derived antimicrobials is the major focus at current to counter antibiotic resistance.This study was conducted to characterize the antimicrobial activity and mode of action of linalyl anthranilate(LNA)against carbapenemase-producing Klebsiella pneumoniae(KPC-KP).LNA alone exhibited bacteri-cidal activity at 2.5％(V/V),and in combination with meropenem(MPM)at 1.25％(V/V).Comparative proteomic analysis showed a significant reduction in the number of cytoplasmic and membrane proteins,indicating membrane damage in LNA-treated KPC-KP cells.Up-regulation of oxidative stress regulator proteins and down-regulation of oxidative stress-sensitive proteins indicated oxidative stress.Zeta po-tential measurement and outer membrane permeability assay revealed that LNA increases both bacterial surface charge and membrane permeability.Ethidium bromide influx/efflux assay showed increased uptake of ethidium bromide in LNA-treated cells,inferring membrane damage.Furthermore,intracel-lular leakage of nucleic acid and proteins was detected upon LNA treatment.Scanning and transmission electron microscopies again revealed the breakage of bacterial membrane and loss of intracellular ma-terials.LNA was found to induce oxidative stress by generating reactive oxygen species(ROS)that initiate lipid peroxidation and damage the bacterial membrane.In conclusion,LNA generates ROS,initiates lipid peroxidation,and damages the bacterial membrane,resulting in intracellular leakage and eventually killing the KPC-KP cells.

Aims: Researchers are focusing more on the isolation of new probiotic bacteria to increase varieties for the growing market demand. This study aimed to isolate lactic acid bacteria (LAB) strains from sugarcane waste materials and evaluate its characteristic. Methodology and results: In the present study, two strains of LAB (Isolates A and B) were isolated from sugarcane waste and investigated in vitro for their characteristics as potential probiotics. These isolates were evaluated on their characteristics based on four biochemical tests (acid tolerance, bile tolerance, microbial adhesion, and phenol resistance), with the commercial strain Lactobacillus isolated from Yakult ® served as a positive control. Both isolated strains (>8 log10 CFU/mL) displayed higher survivability than control (>6 log10 CFU/mL) in simulated gastrointestinal conditions at pH 2.0 and pH 6.9 after 24 h. Furthermore, both isolated LABs were resistant to inhibitory substances which are 0.05-0.3% bile and 0.4% phenol. For bile tolerance, isolate A (OD 6.83) had a higher absorbance at 0.3% bile concentration as compared to isolate B (OD 2.20). However, isolate B (7.49 log10 CFU/mL) showed higher resistance towards 0.4% phenol than isolate A (7.11 log10 CFU/mL) after 24 h. Both isolate A and isolate B displayed low cell surface hydrophobicity, strong electron donor, and basic characteristic. Conclusion, significance and impact of study Both isolates were able to survive under gastrointestinal stress conditions, implying their potential as probiotics. This study demonstrated that valuable products such as probiotic strain could be isolated from sugarcane wastes to use in food production or medical treatment.
Lactobacillales ; Waste Management