Adaptation of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) colorimetric assay for the analysis of Lactobacillus strains
Keywords:
MTT, Lactobacillus, enzyme activity, colorimetryAbstract
Lactic acid bacteria, including Lactobacillus strains, are widely found in nature and are important members of the human intestinal microflora. There has been a lot of research into the beneficial properties of these microorganisms for us, and it is becoming increasingly clear that without them, life would be unimaginable. This is why it is important to have as much information as possible about these essential micro-organisms. In our work, we have adapted and optimized a colorimetric method, originally suitable for human cells, to measure the enzyme activity of the test microorganisms we are investigating and to provide information on the importance of these microorganisms. By modifying and adjusting the parameters of the method reported in the literature, we obtained a rapid method for the direct detection of the enzyme activity of bacterial cells and, indirectly, their cell number in less than four hours. During the adaptation, the optimal parameters (bacterial cell concentration, MTT concentration, incubation time, effect of pH) were established, for which there is a close correlation between cell number and the formazan concentration produced. Our measurements have shown that the enzyme activity of Lactobacillus strains is strongly influenced by their instantaneous growth phase, the quality of the medium and that there is a significant difference between strains.
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