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Bioluciferase Lab Report


We want to find out how enzymes affect chemical reactions in the production of ATP. ATP is an adenine-containing nucleoside triphosphate that releases free energy when its phosphate bonds are hydrolyzed. This energy is used to drive endergonic reactions in cells. In our experiment we will utilize ATP as an energy source to facilitate the conversion of luciferin to its oxidized form by luciferase. We will measure the nucleotide specificity of the luciferase enzyme and how it is influenced by pH changes. We will also measure how different reagents affect the luciferin and luciferase mixture, called lantern. The production of ATP in a biological system requires numerous biochemical reactions that are mediated by various enzymes. An enzyme is a protein serving as a catalyst, which is a chemical agent that changes the rate of a reaction with out being consumed by the reaction. These proteins have a specific three-dimensional shape that includes an active site that fits its specific substrate, which is simply what makes the enzyme work. In this experiment the objective is to use what we know about the release energy in the form of light from the ATP depending on the substrates added. Theoretically, when luciferin mixes with the oxygen in the air and it joins to the active site of luciferase, there should be no reaction because luciferase requires 2 substrates to activate and release energy. When ATP is added, however, it binds with luciferase and breaks off one phosphate group of ATP, creating adenosine diphosphate (ADP). This will release much energy in the form of light that will be visible in a dark room. The release of energy is due to the first law of thermodynamics, which states that energy cannot be created or destroyed, it can only take on different forms. Here, some of the energy will go to make oxidized luciferin, some will come off as heat, and most is released as light.


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