Protein Purification - Biochemistry Lab Extraction...

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Biochemistry Lab 11/18/05 Objectives : During these three weeks, we extracted peroxidase from the seedlings of pea plants and attempted to purify this enzyme. We were aiming at complete purification and to determine how efficient our method of purification is. Peroxidase enzymes are found in nearly every biological tissue. Peroxidase is an enzyme which catalyzes oxidation- reduction reactions. It belongs to a family of enzymes called oxidoreductases and given the enzyme classification number of 1.11.1. There are a few reasons why peroxidase enzymes make a good study for this lab. Firstly, it is very common. Secondly, the enzyme is fairly stable compared to others, so we can work with and handle this enzyme with minimal denaturation. Thirdly, we can visualize the reaction through an assay with chromogenic dyes, and the color change can be measure in a spectrophotometer. Data: Week One Dilution Factor Diltution Factor Zero Initial After 1 minute Change in Abs Activity Activity/gFwt. Crude 1 0.000 0.252 0.482 0.230 0.025556 0.005 2 0.000 0.090 0.275 0.185 0.020556 0.004 4 0.000 0.031 0.074 0.043 0.004778 0.00095 P1 (40%) 1 0.000 0.336 0.422 0.086 0.009556 0.002 2 0.000 0.169 0.217 0.048 0.005333 0.001 4 0.000 0.084 0.103 0.019 0.002111 0.0004 P2 (80%) 1 0.000 0.173 0.621
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This lab report was uploaded on 04/05/2008 for the course BIOL 4177 taught by Professor Pike during the Spring '08 term at East Tennessee State University.

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Protein Purification - Biochemistry Lab Extraction...

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