Enzyme Test Bio 1361 Cheeks

specific activity (enzyme)

The activity of an enzyme per milligram of total protein (expressed in ìmol/time/mg of all protein).

Addition of which metal ion or combination of ions restores alkaline phosphatase activity closest to that of native enzyme?

The addition of Mg, Zn gets the specific activity closest to that of the native enzyme

What reaction does alkaline phosphatase catalyze? Identify the substrate (type of molecule), any other reactants and the product(s).

The enzyme alkaline phosphatase has the ability to catalyze different ranges of phosphate esters, specifically it can be used in numerous reactions such as the hydrolysis of monoesters of phosphoric acid. The enzyme can also be used to dephosphorylate certain compounds so that these molecules can freely diffuse into the cell.

Which ion is least likely to act as a co-factor of native alkaline phosphatase? Explain your reasoning.

The ion least likely to act as a co-factor is Ca since it has the lowest specific activity out of all of them .

What is the maximum specific activity measured for this enzyme and at what pH does it occur?

The maximum specific activity measured for the enzyme is 0.325 umol/ min mg and this occurs at pH 9.5.

What effect did removal of all metal ions have compared to native alkaline phosphatase activity?

The native ALP had a much higher specific activity than all the other metals except for the addition of Cobalt.

Based on your answer to Question 5, predict the pH range at which this enzyme functions in its natural environment.

The pH range the enzyme functions in its natural environment is probably 8.5-10

Over what pH range does the enzyme have 50 - 100% (from half of the y-axis up) of its maximum specific activity?

The pH range the enzyme has 50-100% is 8.5- 10.

Some of the metal ions tested have a positive effect on enzyme activity. What are these types of positive regulators called?

co-factors

In the metal ion experiment, which condition is the control and which condition could be considered a negative control?

control was the native AP negative control is the metal-free AP without any metal added.

Metalloproteins

proteins complexed around a metal ion

Predict the temperature range at which the bacteria live.

40 - 80 C

Over what range of temperatures does the enzyme have 50 - 100% of its maximum specific activity?

60 - 80 C

At what temperature does maximum specific activity occur?

80 degree C

What organisms make alkaline phosphatase enzymes?Do some additional research to answer this question

Bacteria makes up alkaline phosphatase enzymes

Using your knowledge of pH effects on protein function, what is likely to happen to this enzyme at pH 4.5?

If the enzyme was put into a pH of 4.5, the specific activity would dramatically decrease since the enzyme will be denatured.

Express the activity of the metal-free enzyme as a percentage of the activity of the native enzyme.

Metal Free AP type of metal None: 5.4% Mg: 60.81% Zn: 39.19% Co: 862.16% Ca: 4.05% Mg, Zn: 77.03%

Explain why the researchers added EDTA to the solution. Use additional references to provide specific information about which metals interact with EDTA.

The researchers added EDTA to inhibit the ALP by removing the metal. EDTA reacts with heavy metals and minerals