20ml of it was then pipetted into a 250 Erlenmeyer flask, followed by setting up a titration system with 0.100M HCl in the burette. As in the case of titration of an acetic acid with sodium hydroxide, a pH meter was used to monitor the pH of the solution as HCl was being added. HCl was added in an interval of 1ml until the pH of the solution was 9. When the solution pH reached 9.5, HCl was added in an increment of 0.2ml until the equivalence was passed. After this, HCl was added in an interval of 1ml until there was minimal change in the solution pH.
Aim The aim of the experiments to be carried out is to determine the kinetic parameters, Km and Vmax, of Alkaline Phosphatase. Theory, Principles and Application of Principles Enzymes are a huge varying group of proteins which are needed to carry out essential metabolic functions in cells. Substrate-specific enzymes, like Alkaline Phosphatase, act as catalysts lowering the needed activation energy to convert the substrate to product. Enzymes are made up of amino-acids and amino-groups have side chains referred to as R-groups. These R-groups have different degrees of protonation at different pH levels – meaning they can carry different charges at different pH levels, these charges together make up the overall charge of the enzyme.
e.g isocitrate dehydrogenase enzyme acts on isocitrate by oxidation followed by decarboylation Isocitrate is converting into alpha-ketoglutarate with the help of isocitrate dehydrogenase. FACTORS AFFECTING THE RATE OF ENZYME ACTION 1.Effect of nzyme concentration The rate of enzyme is directly propotional to the concentration of enzyme provided that the condition of the reaction remains constant and sufficient substrate is supplied. 2. Effect of substrate concentration The rateof reaction increases as the substrate concentration incteases until a certain point (Vmax) at which the reaction attains maximal velocity. Any Increase in substrate concentration after this point cause further increase in the rate of reaction because at Vmax enzyme moleclues are completed saturated with substrate molecules.
Decomposition of Aspirin Studied with UV/Visible Absorption Spectroscopy Aims: To determine the concentration of salicylic acid, formed from the hydrolysis of Aspirin, at regular intervals using the UV/Visible Absorption Spectroscopy From the concentration of salicylic acid, concentration of Aspirin to be determined using an equation Calculate the rate constant of this reaction and its order from a plot of graph of ln(aspirin) vs time Discuss the overall flaws and improvements to the experiment Results: As per schedule1, 0.212g of aspirin was added to 50 ml boiling water to form salicylic acid in a 100 ml flask, of which 1 ml was then pipetted to a 50 ml volumetric flask at the 5th min. Following an ice bath, the solution was mixed
Introduction: What are enzymes? Chemical reactions that take place in living cells are known as metabolic reactions. There are two types of reactions: • Anabolic Reaction (Constructive) • Catabolic Reaction (Destructive) Substance that accelerate chemical reactions are known as catalysts. Enzymes are biological catalysts, because they accelerate chemical reactions that occur in cells. Activation energy is required to start a chemical reaction.
Experiments have been conducted using optimum values of enzyme loading and ethanol to hexanoic acid molar ratio (as determined from the statistical experimental design) at these three reaction temperatures. Figure 6 shows the Arrhenius Plot of ln (k) versus reciprocal absolute temperature (1/T) from which activation energy can be determined. The thermodynamic parameters for enzymatic esterification were determined using the Eyring equation as follows 20 : ln〖k/T= 〗 [(-ΔH)/R][1/T]+ln[k_b/h ]+ΔS/R [5] ΔH=Ea-RT [6] ΔG= ΔH-TΔS [7] Where h = Planck’s constant (6.626×10-34 J.s), kb = Boltzmann constant (1.381×10-23 J/K), k = rate constant at temperature T, ΔH = Enthalpy of activation, ΔS = Entropy of activation, R = universal gas constant, and ΔG = Gibbs free energy. The values of activation energy and all thermodynamic parameters are given in Table.5. The values obtained from Arrhenius analysis can be substituted in the above set of equations to yield thermodynamic parameters, viz.
Analysis is performed when equilibrium is obtained. In this circumstance, it refers to the association rate and dissociation rate of complex formation being equal. The fractional occupancy at equilibrium is predicted by the law of mass action Firstly, a protein standard curve will be created to determine the concentration of the unknown sample. When this is completed, the saturation binding graph will be created using graph pad prism and determine the Bmax and Kd values. A Scathard plot will then be created and the Bmax obtained from the x-axis intercept and the Kd value obtained from the slope of the line the.
The activation energy must be overcome before the flow process occurs (Rao, 1999). To determine the activation energy at a concentration of 1% w/v and shear rate (100 s-1), the apparent viscosity were studied at different temperatures using the Arrhenius law. The effect of temperature on viscosity of exopolysaccharide was determined by using the following Arrhenius model (Macosko, 1994; Speers and Tung, 1986) η= A.e ∆E/RT ………………………………Equation (4) By taking the logarithm of Eq. (4), a plot of log10 (η) versus 1/T should be a straight line with intercept and slope equal to log10 (A) and Ea/2.303R respectively. A Linear relationship is obtained between log10 (η) and 1/T (Fig 2) with R2 value 0.991.
1. Introduction 1.1 Objective of the Research: The Objective of this research is twofold, first to quantify the information content of the cyclic voltammogram data and understand the kinetic parameters that are involved in redox reaction and simulate them using acquired information, also identify the post oxidation peak that occurs at slow scan rates in the presence of air. Second, to check the electrochemical reduction of nitrate ion at the surface of different working electrodes that are available in the lab at neutral and alkaline systems. 1.2 Importance of the Research: Ferrocene carboxylic acid, as an iron centered inorganic derivative of ferrocene, behaves as a simple and ideal reversible redox system and is commonly used as mediator with enzymes such as glucose oxidase. , , However, during extensive use of Ferrocene Carboxylic Acid, FCA, in this research study,
Alkaline phosphatases (ALP), members of the phosphomonoesterase family, hydrolyze the oxygen-phosphorus bond of organophosphates using metal ions to release an inorganic phosphate under alkaline conditions.1,2 These enzymes are dimeric metalloenzymes containing two Zn2+, one Mg2+, and a serine residue in the active site of each monomeric subunit, in both prokaryotes and higher eukaryotes.2,3 Studies have shown that the three divalent cations are essential for enzymatic activity to catalyze the formation of an alcohol and an inorganic phosphate (Figure 1). In E. coli, the zinc ions are positioned to activate the serine and water for nucleophilic attack and ultimate cleavage of the bond, in addition to holding the phosphate moiety of the substrate. The magnesium ion has been suggested to stabilize the transferred phosphoryl group by a water molecule, using a separate mechanism by which the zinc ions function.3 Although ALPs are found widely in nature and function similarly on the biochemical level, there are locational