Please use this identifier to cite or link to this item: http://hdl.handle.net/20.500.12188/6568
Title: Optimization of enzymatic assay of superoxid dismutase and glutation peroxidase activity in milk whey
Authors: Trajchev, M., Nakov, D., Gjorgoski, I.
Keywords: dairy cows, milk whey, superoxid dismutase, glutathion peroxidase
Issue Date: Oct-2015
Publisher: Faculty of Agricultural Science and Food
Source: Trajchev, M., Nakov, D., Gjorgoski, I. 2016. Optimization of enzymatic assay of superoxid dismutase and glutation peroxidase activity in milk whey. Symposium Proceedings of the 2nd International Symposium for Agriculture and food, 7-9 October, 2015, Ohrid, Republic of Macedonia, pg. 43-51.
Conference: The 2nd International Symposium for Agriculture and food, 7-9 October, 2015, Ohrid, Republic of Macedonia
Abstract: General spectrofotometric kinetic protocols are described to measure the antioxidant enzyme activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in milk whey. The SOD convert superoxide radical into hydrogen peroxide and molecular oxygen, while the GPx convert hydrogen peroxide into water. In this way, two toxic species, superoxide radical and hydrogen peroxide, are converted to the harmless product water. The applicability of this methods for milk samples from dairy cows were tested. SOD activities in the milk whey was determined by the autoxidation of pyrogallol in presence of DTPA in TRIS-HCl buffer, pH= 8.5. The effect of reaction conditions on enzymatic inhibition of pyrogallol autooxidation was studied when different amounts of sample were added in reaction mixture. The reaction mixture for determination of GPx activity contained EDTA, reduced glutathione, glutathione reductase, NADPH and cumene hydroperoxide in phosphate buffer, pH= 7.6. The effect of reaction conditions on enzymatic NADPH consumption was studied when different amounts of reduced glutathione and whey sample were added in reaction mixture. The both spectrophotometric methods were modified in order to accommodate kinetic analyses in 96-well micro plates. The rate of pyrogallol autooxidation and NADPH oxidation linearly depend from amount of whey sample in reactive mixture. Applying higher amounts of milk whey in reactive mixture was decreasing optical density reading of pyrogallol autooxidation at 415 nm and increasing the optical density readings of NADPH oxidation at 340 nm, in a linear way. The methods could be validly applied for milk whey samples.
URI: http://hdl.handle.net/20.500.12188/6568
Appears in Collections:Faculty of Agricultural Sciences and Food: Conference papers

Files in This Item:
File Description SizeFormat 
Optimization of enzymatic assay of SOD and GPx activity in milk whey.pdf219.53 kBAdobe PDFThumbnail
View/Open
Show full item record

Page view(s)

46
checked on Aug 6, 2022

Download(s)

138
checked on Aug 6, 2022

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.