Enzymology Laboratory Lab

Enzymology Laboratory Lab - Nurul Nadzirah Roslan 007544407...

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Nurul Nadzirah Roslan 007544407 Enzymology Laboratory Report Introduction: i. Enzymes are catalysts that accelerate the rates of biological reactions. Enzymes are important to biochemistry because enzymes accelerate the reaction rates many orders of magnitude and, by selecting the substances undergoing reaction, determine the specific reaction taking place, these results that we will be able to maintain the living state. This is because other common ways for chemist to accelerate reactions are not available to cells, such as the temperature cannot be raised, acid or base cannot be added, the pressure cannot be elevated or the concentrations cannot be dramatically increased. ii. NAD + + ethanol acetaldehyde + H + + NADH iii. Environmental conditions affecting the enzymes ability to catalyze the reaction that were tested in this lab: a. pH of pyrophosphate buffer b. concentration of substrate which is ethanol c. concentration of ADH enzyme d. temperature of ADH enzyme iv. The standard assay protocol is start by adding 2.2 mL distilled H 2 O into a cuvette, and then 0.5 mL pyrophosphate buffer, followed by 0.1 mL ethanol and lastly is 0.1 mL of ADH. The mixture solutions is then covered with parafilm and mixed by inverting the cuvette 2-3 times. The cuvette then is placed into the spectrometer and the lid is closed. The wavelength of the spectrometer is made sure set at 340 nm and then the spectrometer is set blank by pressing the “0 Abs” button. After the absorbance reading is displayed 0, the cuvette is took out immediately and then 0.1 mL NAD + is added, the timer is begin simultaneously as the adding. The solution is again mixed by inverting the cuvette 2-3 times before placing inside the spectrometer. The absorbance then is read at every 15 seconds up to 75 seconds. 1
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v. The wavelength 340 nm is chosen because the maximum wavelength for the reduced form of NAD + which is NADH, is 340 nm. Reproducibility: Experiment 2 data and analysis: i. Reproducibility is defined as ability to cause to exist again or to present again or more specifically as the ability to duplicate measurements over long periods of time by different laboratories . ii.
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This note was uploaded on 03/10/2008 for the course CHM 327 taught by Professor Jay during the Fall '08 term at Cal Poly Pomona.

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Enzymology Laboratory Lab - Nurul Nadzirah Roslan 007544407...

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