The Ability of Probiotic Lactobacillus Strains in Removal of Benzo[a]pyrene: a Response Surface Methodology Study

In this study, the ability of various lactic acid bacteria was assessed in removing benzo[a]pyrene (BaP) from contaminated phosphate buffer saline (PBS). Response surface methodology (RSM) was performed using Box-Behnken design to investigate the effect of four independent variables including pH (5-7), incubation time (1-24 h), cell density (10⁷-10⁹ cfu/mL), and initial BaP concentration (5-15 mg/kg) at three levels to evaluate in vitro removal of BaP as response. The results showed that all the tested strains were able to remove BaP from PBS and this reduction was entirely strain-specific. Bifidobacterium lactis BB-12 followed by Lactobacillus casei TD10 exhibited the lowest binding ability while the highest binding rate was related to Lactobacillus acidophilus LA-5, Lactobacillus delbrueckii subsp. bulgaricus PTCC 1737, Lactobacillus casei TD4, and Lactobacillus brevis TD3, respectively. Cyclohexane washing weakened BaP-bacteria complex, while this complex was not significantly changed by PBS washing. The results showed that BaP binding rate was influenced by pH, cell density, time, and BaP concentration in linear and quadratic manners. Moreover, there were interactions between cell density and time as well as between time and BaP concentration. The highest BaP-binding rate by L. acidophilus LA-5 was 10 ppm of BaP concentration, pH = 5, cell density of 10⁹ cfu/mL, and an incubation period of 24 h. It can be concluded that a range of pH, time, and microbial population is required to obtain maximum binding efficiency for BaP based on the concentration of the toxin and the species of the bacteria.