Abstract:
Medicinal plants are considered rich source of ingredients, known to be a potential source of therapeutics or curative aids. The plant product or natural products have an Abiotic conditions have a significant negative impact on the growth, maintenance, and ultimately yield of the plant, which causes significant financial losses and a food crisis. The excessive hazardous reactive oxygen species (ROS) cause oxidative stress. As a result, plants suffer cellular harm and molecular deterioration, which ultimately results in plant death. Plants fight oxidative stress through enzymatic and non-enzymatic mechanisms. A major enzyme of the enzymatic antioxidant system, glutathione reductase (GR) is essential for maintaining the sulfhydryl (-SH) group. It maintains the GSH depletion through the glutathione ascorbate pathway. Optimization of colorimetric assay for Glutathione reductase was assessed by varying different parameters: ?max, effect of different concentrations of substrate, effect of incubation time, volume of crude extract concentration. The methodology was based on the rise in absorbance at 412 nm that occurs when GSH reduces 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB). The specificity of DTNB in particular makes it possible to test for glutathione reductase even when there are other NADPH-dependent enzymes are present in crude protein extract. In contrast to NADPH, GSSG serves to protect the enzyme against inhibition. For enzyme specificity kinetic study was performed in which the Km value for substrate (GSSG) was 0.145 M and Vmax was 8.93 M/sec. Higher Vmax and lower Km for GSSG (substrate) were observed and literature provide a reason that it is due to decrease in intracellular GSH that act as noncompetitive inhibitor of glutathione reductase on the other hand result also showed an ordered sequential mechanism that shows that despite of low GSH, glutathione reductase forms strong complex with GSSG
Page(s):
86-86
DOI:
DOI not available
Published:
Journal: Abstract Book on Global Science Technology and Management Conference, Volume: 0, Issue: 0, Year: 2023
Keywords:
Cholistan desert
,
Glutathione
,
ROS
,
spectrophotometers
,
Glutathione reductase