Investigation of biochemical and antioxidant effects of rosmarinic and vanilic acids against glyosate toxicity in vitro medium

Investigation of Biochemical and Antioxidant Effects of Rosmarinic and Vanilic Acids Against Glyphosate ToxicityIn Vitro Medium Glyphosate inhibits the growth of plants by inhibiting the activity of enzyme 5-enolpyruvyl-shikimic-3-phosphate synthase (EPSPS), which is required for the formation of aromatic amino acids such as tyrosine, tryptophan and phenylalanine. Rosmarinic acid and vanillic acid are naturally occurring phenolic acids in many plants. In this study, Saccharomyces cerevisiaeL. was used as a model organism in studies in molecular biology and biochemistry and antioxidant and biochemical effects of rosmarinic and vanillic acid added at different doses to the culture medium against the toxic effect of glyphosate were investigated. Both glyphosate and rosmarinic and vanillic acid concentrations in culture media were added as 200, 400 and 800 mg per liter. The culture broth was autoclaved at 121 ° C for 15 minutes. After cooling at room temperature, yeast cells were inoculated in sterile medium. After this procedure, the 30 ºC sample was allowed to incubate for 72 hours. At the end of the period, the concentrations of the culture fluids were measured at 600 nm in the spectrophotometer. The cell pellet was then isolated by centrifugation at 6000 rpm at +4 ° C.The cell pellet was washed with 0.9% NaCl solution and then homogenized with Tris-EDTA buffer (pH = 7.4) and the upper supernatant was separated from the pellet at 6000 rpm at +4 º C. Total protein, GSH, GSSG, MDA and Glutathione S transferase activities were measured from the supernatant.The remaining pellet portion was homogenized with a mixture of 3/2 (v / v) n-Hexane / isopropyl alcohol. The supernatant fraction was removed by centrifugation and esterification was performed to analyze fatty acids in gas chromatography. Analysis by gas chromatography revealed that the yeast cell pellet extract contained octanoic (8: 0), lauric (12: 0), myristic (14: 0), palmitic (16: 0), palmitoleic (16:1 n-7), stearic (18: 0), oleic (18:1 n-9) and linoleic (18:2 n-6) acids were observed. Analyzes of vitamin A, D, E, and K vitamins were performed with an HPLC instrument. As a result of this analysis, molecules such as K2, δ-tocopherol, D2, D3, -tocopherol, ergosterol, K1, stigmasterol, -sitosterol were identified. Biochemical analyzes indicated that cell density decreased in culture medium containing 800 mg glyphosate. Protein amount, GST activity, lipid peroxidation and fatty acid levels were observed. . It was determined that vanillic and rosmarinic acid against glyphosate toxicity were effective on biochemical values in Saccharomyces cerevisiaeL. culture medium. Key words: Glyphosate, Vanilic acid, Rosmarinic acid, Antioxidant effect, Total protein, GSH, GSSG, Lipid peroxidation, Fatty acid, DPPH, GST, Saccharomyces cerevisiaeL.