Enzyme kinetics

Malachite Green Assay: The reaction that AGPase catalyzes produces PPi. The resulting PPi that is produced is hydrolyzed by an inorganic pyrophosphatase (Sigma Aldrich) to generate phosphate (Pi). Each reaction for the assay was conducted on a 96 well plate and the reaction mixture was 50µL and contained Pre-Mix which contained 50mM HEPPS, pH=8.0 (MP Biomedicals), 10mM MgCl2, 1.5mM FBP (Sigma Aldrich), and 0.2mg/mL Bovine Serum Albumin (Sigma Aldrich), and 1.5mU/µL yeast inorganic…

Alkaline phosphatase catalyzes the hydrolysis and transphosphorylation of phosphomonoesters (1). A kinetic reaction scheme of AP is presented in Figure 2. Figure 2: Kinetic reaction scheme of alkaline phosphatase (11). Under the hydrolysis reaction, free phosphate and an alcohol molecule are released from the active site. The rate-determining step of this reaction is pH dependent. Transphosphorylation has similar initiate steps as hydrolysis. However, phosphate cannot exit the active site,…

at which enzyme activity is most effective is approximately at 40℃. This result was expected as this trial temperature was closest to internal body temperature, 38℃, and as catalase is present in all body organs (T. Fouad, Antioxidants). Temperature affects chemical reactions by increasing the kinetic motion of molecules and changing the rate of collisions between them. In lower temperatures, kinetic motion is decreased and there are not enough collisions between the molecules for enzyme…

As macromolecular machines, enzymes conduct their designated work in the confined and crowded spaces inside living cells. How does a protein work in such a condition? Functions of proteins are strongly dependent on their structural stability and dynamics while the (macro) molecular interactions are arguably affected by the complexity and diversity of their environments. To date, two interrelated cellular conditions — ‘macromolecular crowding’ resulted from the effect of volume exclusion by…

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…

Introduction An enzyme is a protein that functions as a catalyst, it speeds up chemical reactions (Freeman et al. 2017 p. 90). Enzymes are often large globular proteins and are able to hold substrates in specific orientations so they are able to react, the location where the substrate binds with the enzyme and reacts is called the active site and is the location where the catalysis occurs (Freeman et al. 2017). Enzymes activity is often related to the optimal environment for them in respect to…

it will result in faster rates of enzyme activity. Therefore, if the opposite were to occur, then the reaction will slow down and not function properly. For concentration, my hypothesis was proved to be correct. The undiluted solution had higher frequencies of collision at the active site of the enzyme and substrate. Which means the percent of amylose as well as absorbance level was higher compared to the diluted samples. The result on the effect of pH on enzyme activity prove my hypothesis to…

and the amount of energy available for the reaction. The reaction rate of any enzymes, including α-Amylase, is directly proportional to the temperature before it reaches a certain limit. As the temperature reaches this limit (which was observed to be 37°C in this experiment) , causing the reaction to also be the speed up the most, it is high enough to denature the protein composition of the enzyme, resulting in the enzyme losing its initial shape and therefore the ability to bind securely to a…

other enzymes studied were more effective for anthraquinone-dye (Reactive Blue19) than azo-dye…

modification of the substrate $S_0$ to $S_1$, with enzyme $E$ and phosphatase $F_1$. $S_1$ acts as an enzyme for the modification of the substrate $P_1$ to $P_2$, and this has a phosphatase $F_2$. The following matrices describe the system, The method draws the conclusion that the family is $S$-injective, using the determinant criterion, in the first iteration Consider the modification of the substrate $S_0$ to $S_1$, with enzyme $E$. $S_1$ acts as an enzyme for the modification of the…