Anti-Candida activity of spent culture filtrate of Lactobacillus plantarum strain LR/14

J Mycol Med. 2014 Jun;24(2):e25-34. doi: 10.1016/j.mycmed.2013.11.001. Epub 2013 Dec 6.

Abstract

Objectives: This study was undertaken to understand the effect of antimicrobial compounds produced by an environmental isolate of lactic acid bacterium, Lactobacillus plantarum strain LR/14, on growth, viability and biofilm forming ability of the pathogenic yeast, Candida albicans SC5314 and to identify the mode of action of such compounds.

Material and methods: L. plantarum LR14 was grown at 37°C for 18 h in MRS broth. The spent culture filtrate (SCF) was collected by centrifugation and checked for anti-Candida activity. Live/dead staining followed by fluorescence microscopy was done to study the membrane damage. Increased membrane permeability was confirmed by measuring the release of ions and macromolecules (ATP) using atomic absorption spectrophotometer and luminometer, respectively. Effect on biofilm formation was quantified by MTT reduction assay.

Results: The viability of yeast cells was affected by SCF LR14 treatment in a dose-dependent manner, exerting a fungicidal effect. The active compound was identified as a pH-dependent thermostable proteinaceous metabolite. The fungicidal activity was further confirmed by PI staining, suggesting compromised membrane as the cause of cell death. Leakage of intracellular contents such as, K+ ions and ATP, as a cause of its inhibitory action further confirmed the membrane disruption. Moreover, significant reduction in biofilm formation was also confirmed.

Conclusions: SCF LR14 showed potent anti-Candida activity, affecting cell viability, membrane permeability, and biofilm formation and leading to cell death, thereby suggested a probable candidate as a natural therapeutic agent.

Keywords: Anti-yeast compound; Bactérie acide lactique; Composé anti-levure; Filtrat de culture; Lactic acid bacteria; Permeability; Perméabilité membranaire; Spent culture filtrate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Infective Agents / isolation & purification
  • Anti-Infective Agents / metabolism
  • Anti-Infective Agents / pharmacology*
  • Biofilms / drug effects
  • Biofilms / growth & development
  • Candida / drug effects*
  • Candida / growth & development
  • Culture Media, Conditioned / chemistry
  • Culture Media, Conditioned / pharmacology
  • Filtration
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Lactic Acid / metabolism
  • Lactic Acid / pharmacology
  • Lactobacillus plantarum / cytology*
  • Lactobacillus plantarum / metabolism
  • Microbial Sensitivity Tests
  • Peptide Hydrolases / metabolism
  • Peptide Hydrolases / pharmacology

Substances

  • Anti-Infective Agents
  • Culture Media, Conditioned
  • Lactic Acid
  • Peptide Hydrolases