Enzyme assay

The hypothesis of the experiment was that the molecular weight of N-acetyl-β-D-hexosaminidase would be around 28,000 kDa but to determine the molecular weight the concept of SDS-PAGE had to be understood. The movement of any charged particle through an electric field is determined by its net charge, its molecular radius and the magnitude of the applied field (7). Being that natively folded proteins are not molecular weight dependent, the proteins would be traveling at different speeds in an…

Introduction Lactate dehydrogenase also known as LDH is a critical enzyme that is found in all mammals. It exists as a tetramer in five isoforms with a combination of heart and muscle subunits. One of its main functions is to catalyze the reaction of pyruvate to lactate. This reaction occurs in an anaerobic environment where the body needs to make sufficient energy as fast as possible, therefore pyruvate ferments to lactate with the help of LDH to produce enough NAD+ to power glycolysis. LDH-5,…

CYP2D6 is a Phase I oxidative metabolism enzyme that is clinically important because about 20-25% of clinically used drug are metabolized by the CYP2D6 enzyme. CYP2D6 substrates are typically lipophilic and include drugs such as antidepressants, antipsychotics, antiarrhythmic, antiemetic, and opioids. The exact clinical effect of the polymorphism is dependent on the drug and the variant alleles expressed because individual allele variants can lead to a range of metabolic ability from ultrarapid…

Sodium taurocholate. In the absence of the pure sodium taurocholate, the activity in the range of pH 4.0-4.8 was considerably lower. However, in the presence of pure Sodium taurocholate, leukocyte GCase was optimally active at pH 5.0, after which the enzyme activity started to decline. Figure 1 also shows that leukocyte glucocerebrosidase activity is completely dependent upon this detergent. Maximum stimulation representing a four- to five-fold increase in activity occurred…

For the first part, the qualitative polyphenol oxidase test (Table 1), we looked at 3 qualitative test tubes, where only test tube 2 turned orange since this tube had both the 40 mM dopa, the substrate and PPO enzyme extract. However, the test tube 3 did have a cloudy component because of the presence of PPO extract, but there was no color change. For a color change to occur, there needs to be a presence of both the dopa and PPO extract. Furthermore, we calculated the final concentrations of…

The ΔglgC cyanobacteria will be transformed yet again to express an engineered new, small PSII antenna. In order to do this, plasmids pGDNG3/Cys(181/226)Ser and p613 will be co-transformed using the method described in the paper by Beckman et al. The method of using a vector containing an antibiotic (ampicillin) resistance gene will also be employed in this experiment. Ampicillin will then be added to the transformed bacteria in order to obtain the successfully transformed bacteria. By adding…

DM and GalA content were determined according to the method reported by Aina et al., [32] and Food Chemical Codex 1996 [33]. 0.1g of pectin was weighed into a 250 mL conical flask, then wetted with 5mL ethanol. 100mLof deionized water was added, then six drops of phenolphthalein (0.1% in ethanol) indicator to each sample. Pectin dispersions were stirred until it was fully dissolved, thereafter, the solution was titrated slowly with 0.1N NaOH until the colour of the indicator just changed to…

liver was manipulated primarily through centrifugation, and mitochondrial fractions were then created to serve as the enzyme-rich solutions the reagent would act upon. To determine the protein concentration of the mitochondrial fraction, the Bradford assay was implemented; the result will later be used in the calculation of the specific activity of SDH. Ultimately, the SDH assay provided the data to either support or negate the hypothesis about malonate’s effect on SDH activity. This procedure…

the accuracy of separation. The purpose of Experiment 5-2 is to purify the E. coli enzyme, β-Galactosidase through the use of affinity chromatography on P-aminobenzyl-1-thio-β-D-galactopyranoside agarose, which is a good substrate for the enzyme. This enzyme will be eluted at high pH. The Bradford assay will be used to determine the protein concentration and the specific activity of the solutions containing the enzyme. Size-exclusion chromatography separates molecules…

the effect of Aerva lanata aqueous suspension reduced the urinary levels of stone promoters such as oxalate, calcium, uric acid, phosphorus and protein and increased the content of stone inhibitors citrate and magnesium. The oxalate synthesising enzymes that is, lactate dehydrogenase and glycolic acid oxidase also lessened in the liver and kidneys. The histopathological studies showed dissolution of calcium oxalate stones compared to disease control…