Bio Chem Experiment

Topics: PH, Enzyme, Concentration, Chemical equilibrium, Temperature, Product / Pages: 13 (3137 words) / Published: Jan 17th, 2014
Enzyme kinetics

Purpose: The goal of this investigation was to measure the amounts of products made and see the different elements that that affect the rate of breakdown of p-Nitro phenol in the absence or presence of cellobiase…..

Methods:
Activity #1
The materials used for this activity are as follows: 1.5 mM substrate, enzyme, Stop solution, buffer, DPTPs, 15 ml conical tubes, cuvettes, marker, beaker, distilled water, spectrophotometer, stop watch.
First four 15 ml conical tubes were labeled: “Stop solution“, “1.5 mM Substrate”, “Enzyme”, and “Buffer”. Then, seven cuvettes were located and five of them were labeled “E1-E5” and the other two were labeled “Start” and “End”. After 500 ul of “Stop solution“ was pipetted using a clean DPTP into the seven cuvettes. Next, two 15ml conical tubes were located and respectively labeled “ Enzyme Reaction” and “ Control”. Immediately, after 2ml of “1.5 mM Substrate” was added to the 15ml conical tube labeled “Enzyme Reaction”. After 1 ml of “1.5 mM Substrate” was pipetted into the conical tube labeled “Control”. Then 500ul of buffer was put into the 15 ml conical tube and labeled control and this was mixed. Immediately, after finishing mixing the solution 500ul was taken out of this and placed into the cuvette labeled “Start”. After, 1 ml of enzyme was pipetted into the 15ml conical tube labeled “Enzyme Reaction “, this conical tube was then mixed. Post mixing, 500 ul of this solution was removed and placed into the cuvette that was labeled “Stop Solution”. A timer was started after this step. Then, at intervals of 1 min, 2 min, 3 min,4 min, 8 min 500ul of the solution from the “Enzyme Reaction “ was removed and placed into the cuvettes labeled “E1-E5”. After the entire enzyme samples were collected 500ul of the solution labeled control was added to the cuvette labeled “End”. The absorbance of each of the cuvette’s was taken and recorded. These were

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