6-Phosphogluconate Lab Report
5. RESULT AND DISCUSSION
RESULT
The enzyme 6-phosphogluconate is the significant enzyme in the oxidation phase of pentose phosphate pathway. The homogenate was made by adding the buffer to the liver of the quail after one hour centrifuge at 12300 RPM materializing at 4°C. Precipitation protein was working by ammonium sulfate; it loaded directly to column 2’, 5’ ADP Sepharose 4B affinity chromatography, which has been a high affinity to NADP+ substrate. The flow rate of elution was 23 ml/hr. Taking 18 fractions, checking activity for each fraction at 340 nm. The activity found in fraction tube six to fourteen. This process was materializing at 4°C for 5-6 hour at least to achieve a better yield 89% result and high activity of enzyme …show more content…
By RF was finding the molecular weight of the enzyme. The molecular weight of 6-PGD is 44 KDa. Studying properties (characterization) of the enzyme while the enzyme is more active in optimum temperature at 60°C, in Tris-HCl PH=8. 0 shown in the figure (4.4.) high activity. The optimum ionic strength was shown in 0.6 M Tris-HCl in figure (4.6.). The kinetic study supply how the reaction occurs in the present of enzyme and substrate. Find Vmax=0. 0078, Km= 0.058 at stable 6-PG, Vmax= 0.057, Km=0. 0015 at stable NADP+. The kinetic study provided how reaction took place and participation of a substrate with an enzyme to get production. The stability of PH in Tris-HCl shown in figure (4.7.) high activity of enzyme 6-PGD at PH=8. 0, stability of PH in K-phosphate shown in figure (4.8.) high activity of enzyme 6-PGD at PH=7. 0 in the period of time (every eight hours) until 32 …show more content…
The antioxidant defense system protects the cells from those molecules by GSH reduced. (Ames et al, 2003; Deleva and Erine Kaplowitz 1991). The lack of G6PD enzyme and 6-PGD enzyme in patients are suffering from hemolytic anemia which leads oxidative stress. Oxidative stress was coming from drugs, chemical exposure, and bad foods. The proteins and other enzymes by free radicals were damaged red cells, which going out in blood circulation to the spleen. The kidney excreted hemoglobin to bilirubin, but this case did in severing patients, resulting from renal failure (Ichikawaet al., 1994; Witko et al.,
RESULT
The enzyme 6-phosphogluconate is the significant enzyme in the oxidation phase of pentose phosphate pathway. The homogenate was made by adding the buffer to the liver of the quail after one hour centrifuge at 12300 RPM materializing at 4°C. Precipitation protein was working by ammonium sulfate; it loaded directly to column 2’, 5’ ADP Sepharose 4B affinity chromatography, which has been a high affinity to NADP+ substrate. The flow rate of elution was 23 ml/hr. Taking 18 fractions, checking activity for each fraction at 340 nm. The activity found in fraction tube six to fourteen. This process was materializing at 4°C for 5-6 hour at least to achieve a better yield 89% result and high activity of enzyme …show more content…
By RF was finding the molecular weight of the enzyme. The molecular weight of 6-PGD is 44 KDa. Studying properties (characterization) of the enzyme while the enzyme is more active in optimum temperature at 60°C, in Tris-HCl PH=8. 0 shown in the figure (4.4.) high activity. The optimum ionic strength was shown in 0.6 M Tris-HCl in figure (4.6.). The kinetic study supply how the reaction occurs in the present of enzyme and substrate. Find Vmax=0. 0078, Km= 0.058 at stable 6-PG, Vmax= 0.057, Km=0. 0015 at stable NADP+. The kinetic study provided how reaction took place and participation of a substrate with an enzyme to get production. The stability of PH in Tris-HCl shown in figure (4.7.) high activity of enzyme 6-PGD at PH=8. 0, stability of PH in K-phosphate shown in figure (4.8.) high activity of enzyme 6-PGD at PH=7. 0 in the period of time (every eight hours) until 32 …show more content…
The antioxidant defense system protects the cells from those molecules by GSH reduced. (Ames et al, 2003; Deleva and Erine Kaplowitz 1991). The lack of G6PD enzyme and 6-PGD enzyme in patients are suffering from hemolytic anemia which leads oxidative stress. Oxidative stress was coming from drugs, chemical exposure, and bad foods. The proteins and other enzymes by free radicals were damaged red cells, which going out in blood circulation to the spleen. The kidney excreted hemoglobin to bilirubin, but this case did in severing patients, resulting from renal failure (Ichikawaet al., 1994; Witko et al.,