Enzyme assay

i. Enzymes are macromolecules that increase chemical reactions without being consumed in the reaction. They do this by lowering the activation energy (the amount of energy that reactants need to absorb in order for a chemical reaction to take place). Every enzyme has a substrate with a unique shape and structure for it to bind to. Denaturation of proteins (when a protein unravels and becomes inactive), and change in the shape of the enzyme or substrate all influence the activity and…

Exploring Enzymes Dmitri N. Piquero Biological Discovery I Laboratory Section I Abstract Amylase is an enzyme that speeds up the conversion rate from starch to glucose. Four test tubes were filled with a starch solution and a different concentration of amylase solution ranging from 0 μL to 1000 μL. The test tubes were then left to soak in a 37°C water bath before having 2 mL of 1 M hydrochloric acid added to a test tube of pure amylase solution. Lugol’s iodine was used to test for the…

energy required. An enzyme is a substance produced by a living organism that acts as a catalyst to bring about a specific biochemical reaction. Enzymes carry out the thousands of chemical reactions that occur on cells, or the basic units of structure and function of an organism. Usually, they are polymers of thousands of amino acids. Enzymes are not used up as it carries out the reaction but are recycled over and over. In this experiment, peroxidase from horseradish was the enzyme used to test…

using a 100uL-1000uL micropipette and micropipette tips. The first was the blank solution that only contained 800 microliters of distilled water and 200 microliters of the BioRad protein assay reagent. The second fraction was made with 200 microliters of distilled water, 600 microliters of the BioRad protein assay reagent, and 200 microliters of Bovine serum albumin standard solution (BSA). The following fractions had the amount of water used decrease by 200 microliters and the amount of BSA…

Characterize the inhibitory activity of thiophene compounds against Pks13-TE using Isothermal Titration Calorimetry Zhang, W et al. has Recently shown that thiophene compounds are expected to form non-covalent interactions with Pks13-TE 18. The 4MH-based assay, which is explained in aim 1 will be used to assess the thiophenes inhibitory activity of each thiophene compound that are received from Dr. Steve Sucheck’s…

Cytotoxicity assay: Cytotoxic and viability assay of pulchranin A on human cancer cell lines: The isolated compound exhibited a moderate cytotoxic effect on the three tested cell lines: HCT, HepG2 and MCF-7 where the cell viability, measured by SRB assay, and the IC50 was found to be 91, 80, 63 µg/ml respectively (Figure 3). The standard used was doxorubicin, which gave an IC50 value…

remains. Enzyme and Substrate Solution Preparation A blank tube was prepared by adding 5 mL 0.08 M phosphate buffer (7.7 pH) and 0.5 mL of 2.5 nM ONPG to a cuvette to zero the spectrophotometer with the spectrophotometer set to 420 nm. The substrate solutions were prepared individually by adding 4.5 mL 0.08 M phosphate buffer (pH 7.7) to a Spec 20 cuvette with 0.5 mL of ONPG. The enzyme solution was prepared according to enzyme isolation process. Enzyme Assay 0.5 mL of the isolated enzyme was…

To successfully co-reconstitute both enzymes into a single liposome it was necessary to first conduct detergent screening experiments for the reconstitution of each enzyme independently. To establish the most suitable detergent and detergent concentration, the magnitude of PMF generation along with enzymatic activity was measured for each reconstituted enzyme. Of the detergents tested, the zwitterionic surfactant CHAPS was found most suitable for…

to NAD+ by CMI(9) . By coupling the associated metabolisms of these two enzymes NADH is produced using light and water as substrates with oxygen as the only…

RNA to proteins. The type of protein created relies on what type of messenger RNA or mRNA that was produced during the transcription of DNA. In our experiment, we looked at the specific LacZ gene that codes for beta-galactosidase, which is a protein/enzyme that can break down lactose into its constituent monosaccharrides, glucose and galactose. The amounts of proteins produced by this gene were observed to see what strain would yield the most. Each of the strains tested had a mutation that would…