Bio Lab 5 Properties of Enzymes PDF

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Duong Duong BIOL 1500 Section D25 Colleen McMaken 15 October 2020 Lab 5: Properties of Enzymes Post-Lab Assignment Part 1: Laboratory Notebook Questions: 1. The enzyme that was used was alkaline phosphataste because it is optimally active at alkaline pH environments, which its hydrolysis will turn the colorless pNPP to yellow-color pNP, which will be used to monitored and used as an index of rate of reaction. 2. The color change was created by the hydrolysis of the phosphate, which making colorless pNPP turing into yellow-color pNP. 3. Alkaline buffer was used for Parts 2 and 3 but not for Part 1 because buffer is a solution that resists changes in pH in addition of a small amount of strong acid or strong base. 4. It is essential to wait to add the enzyme because solution D has enzyme with high concentration. In part 1, the pH optimal was estimated for enzyme ALP activity, so we have to wait in order for the cuvettes in spectrophotometer to be accurate. 5. It is essential to incubate the enzyme for at least 20 minutes in Part 3 because it is essential to determine the effect of temperature on catalytic rate. The longer it is incubated the better. If they are not incubated enough, the cuvettes will return to room temperature, which will affect the result.

6. It would be a source of error to leave the sample in the spectrophotometer the entire time because the spectrophotometer is a lightsource, leaving the enzyme sample in there will introduce heat, which is not the desired variable for concentration or pH. Therefore, leaving the sample in in spectrophotometer will alter the temperature which yield the wrong data.

Part 2: Results: Table 1: Average Enzyme Activity Enzyme Activity (U) pH Acidic (4) Neutral (7) Basic (10)

Average -0.001488333 0.0013625 -0.001125833

SD 0.00286857 0.00233276 0.00228556

Figure 1.1: Average Enzyme Activity of ALP for different pH values.

Figure 1.2: Average Enzyme Activity of ALP for different pH values.

Table 2: Enzyme Activity in Different Enzyme Concentrations Enzyme Activity (U) Concentration Average SD Low 0.00026833 0.00012173 Medium 0.00077333 0.00028477 Medium-High 0.00236583 0.00183326 High 0.00187667 0.0012751

Figure 2.1: Average Enzyme Activity in Different Enzyme Concentrations.

Figure 2.2: Average Enzyme Activity in Different Enzyme Concentrations.

Table 3: Average Enzyme Activity in Different Temerature Enzyme Activity (U) Temperature Average SD 4°C 0.00011 0.00013902 23°C 0.000220833 0.00011519 32°C 0.0002575 0.00016457 60°C 0.000190833 0.00012227

Figure 3.1: Average Enzyme Activity in Different Temperatures

Figure 3.2: Average Enzyme Activity in Different Temperatures...