ANTIOXIDANT PROPERTIES OF BIFIDOBACTERIUM BIFIDUM AND LACTOBACILLUS REUTERI CELL-FREE EXTRACTS IN VITRO
About the author:
Knysh O. V., Nikitchenko Yu. V.
Type of article:
Some probiotic bacteria possess antioxidant activity (AOA). This is due to their ability to chelate metal ions, produce their own antioxidant enzymes and metabolites, enhance AOA and increase the level of antioxidant metabolites in the host, regulate signaling pathways, suppress the activity of enzymes responsible for the production of reactive oxygen species and regulate the composition and activity of the intestinal microbiota. However, the realization of the beneficial effects of probiotics requires colonization of mucous membranes by them and engraftment in the gastrointestinal tract. Low levels of engraftment and insufficient safety of cellular probiotics require the use of alternative approaches to probiotic therapy. One is the use of biotechnological products based on biologically active derivatives of probiotics. Our method of obtaining cell-free extracts (CFEs) from probiotic bacteria allows obtaining in the final product a mixture of structural components and metabolites of probiotics without impurities of a nutrient medium. Their biological properties need careful study. In our previous studies Bifidobacterium bifidum and Lactobacillus reuteri CFEs showed antiradical activity with respect to the most reactive hydroxyl radical which exceeded the same activity of the metabiotic Hylak® Forte. The aim of the study was to investigate the antioxidant, glutathione peroxidase (GPx) and catalase (CAT) activity of CFEs obtained from disintegrates and cultures of B. bifidum and L. reuteri cultured in their own disintegrates with or without supplementation. Methods. CFEs were obtained by disintegration of L. reuteri and B. bifidum probiotic strains (L and B extracts) or by cultivation of these strains in their own disintegrates without supplementation (ML and MB extracts) and supplemented with 73,7 g/l glycerol and 72,1 g/l glucose (MLG extract). Metabiotic Hylak® Forte (HF, Ratiopharm, Germany) and the well-known antioxidant α-tocopherol (ICN, USA) were selected as the comparative preparations. The AOA of CFEs, metabiotic HF and α-tocopherol in the model system with yolk lipoproteins as well as CAT- and GPx-activity of CFEs and metabiotic HF were investigated by spectrophotometric method using double beam spectrophotometer «Specord UV VIS» (Jena, Germany). Results. It was found out that CFEs from disintegrate (L) and culture of lactobacilli (MLG), cultivated in its own disintegrate supplemented with glycerol (73.7 g/l) and glucose (72.1 g/l) have significant AOA. They provided 50% inhibition of the induced lipid peroxidation (LP) intensity at a concentration of 1.4 mg/ml (L) and 2.4 mg/ml (MLG) in the reaction medium. α-Tocopherol and metabiotic HF reduced the induced LP intensity in the model system by 50% when contained in the reaction medium 8 mg/ml and 7.4 mg/ml, respectively. Extract B at a concentration of 17 %vol. (1.6 mg/ml) in the reaction medium provided a decrease of the induced LP intensity by only 16.2 ± 8 %. The induced LP intensity in the model system at content of MB extract 17 %vol. (1.5 mg/ml) did not significantly differ from the corresponding indicator in the control sample (without CFEs). Neither the CAT- nor GPx-activity of the studied CFEs and the metabiotic HF were detected at their content in the reaction medium up to 200 μl/ml. Conclusion. The AOA of the CFEs from the L. reuteri disintegrate (L) and culture, cultivated in its own disintegrate supplemented with glycerol and glucose (MLG), corresponded to the level of AOA of the well-known antioxidant α-tocopherol and the metabiotic HF. Extracts from B. bifidum disintegrate (B) and cultures of B. bifidum and L. reuteri cultivated in their own disintegrates (MB and ML, respectively) had low or insignificant AOA. Under the experimental conditions none of the CFEs and metabiotic HF showed CAT- and GPx-activity.
probiotics, cell-free extracts (CFEs), induced lipid peroxidation (LP), antioxidant activity (AOA).
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Publication of the article:
«Bulletin of problems biology and medicine» Issue 2 (156), 2020 year, 236-240 pages, index UDK 615.372:[579.864.1+579.873.13]:577.352.38