Enzymes make up large and diverse groups of biologically active proteins that are essential for metabolic function in cells. They are very specific and only work on special molecules known as substrate. They form an enzyme-substrate complex that serve as catalyst with the ability to speed up chemical reactions that would otherwise take place too slowly for the body to benefit; enzymes provide an alternative reaction pathway of lower activation energy. Therefore understanding the kinetic behaviour of an enzyme provides clues to its possible physiological role1. The purpose of this experiment is to study the mechanism of enzyme-catalyzed reaction in order to determine its effects with changes in a) enzyme concentration, b) substrate concentrations c) inhibitor concentrations d) PH and e) Temperature. Enzymes are composed of residues that have their own characteristics. The degree to which those side chains are protonated by changes in PH will affect the formation of the enzyme-substrate complex as those side chains are involved in many interactions for example …show more content…
Alkaline phosphatase is a hydrolase present in all tissues throughout the human body, particularly in the liver, bile duct, and kidneys and in the placenta. Its function is to hydrolyse phosphor-ester bonds, releasing a phosphate group and the alcohol derivative of the substrate2, 3. A kinetic assay is performed using the substrate P-Nitro phenol-phosphate for the synthesis of its product P-Nitro phenol that is phosphate-free and highly colored. Therefore this provides the possibility of carrying a spectrophotometric enzymatic assay. When alkaline phosphatase (enzyme) acts in p-nitro phenol phosphate (substrate), it forms p-nitro phenol (product), which is a yellow solution that has a high molar absorptivity at 400 nm. The concentration of the product (p-nitrophenol) is then determined by using the beer Lambert