Grossmont College Cellobiases Enzymatic Activity Discussion

November 28, 2020| Homework Lance

Grossmont College Cellobiases Enzymatic Activity Discussion

Skill Check Worksheet Exercise 9:                   NAME_____________________

 Introduction to Enzyme Reactions                   Lab Section______________     

Activity 1: Determining the Reaction Rate in the Presence or Absence of Cellobiase

 

Data Table 1: Comparison of Reaction Cuvettes to Standard Cuvettes.

 

Time (minutes) Cuvette Standard that is Most Similar Amount of p-nitrophenol (nmol)
0 START S1  
8 END S1  
1 E1 S2  
2 E2 S3  
4 E3 S4  
6 E4 S5  
8 E5 S5  

 

 

Cuvettes after mixing enzyme sample with stop solution

 

 

Calculation of Initial Rate of Product Formation (with enzyme present)

 

 

 

 

 

 

Calculation of rate of Product Formation with no enzyme present

 

 

 

 

 

 

Activity 1 Questions

 

 

  1. Describe any changes you observed in the enzyme reaction and control reaction conical tubes during the time that the reaction was occurring?
  1. What happened to the solution in each cuvette after you added the enzyme/substrate mixture to the stop solution that was in the cuvette?
  1. Describe the amount of product produced in the enzyme-catalyzed reaction compared to the control where no enzyme was added.
  1. If you took another time point at 15 minutes, do you think more product would be produced than at 8 minutes? Explain your answer.

 

  1. Is the rate of product formation constant over time? (Hint: Is the slope of the line constant or does it change over time?)

BE SURE TO ATTACH YOUR GRAPH FOR THIS ACTIVITY TO THE SKILL CHECK WORKSHEET WHEN YOU TURN IT IN!

Activity 2: Determining Effects of Temperature on Cellobiase Activity

 

Data Table 2: Determination of ρ-nitrophenol produced at three different temperatures based on ρ-nitrophenol standards

Temperature Standard that is most similar Amount of ρ-Nitrophenol Produced (nmol)
0C S2
Room temp S3
37C S5

BE SURE TO ATTACH YOUR GRAPH FOR THIS ACTIVITY TO THE SKILL CHECK WORKSHEET WHEN YOU TURN IT IN!

 

At what temperature does this enzyme work best? Explain how you know this.

 

 

 

 

Why do chemical reactions occur faster at higher temperatures? Is that true for all enzymes, and unlimited temperature values?

 

 

 

Why do most enzymatic reactions slow down at extremely high temperatures?

Activity 3: Determining Effects of pH on Cellobiase Activity

 

 

Data Table 3: Determination of ρ-nitrophenol produced at three different pH’s based on p-nitrophenol standard

pH Standard that is most similar Amount of p-Ntrophenol Produced (nmol)
5.0 S3
6.3 S2
8.6 S1

Activity 3 Questions

  1. At what pH does this enzyme work best? Explain how you know.

 

 

 

 

 

 

 

 

 

  1. Why do most enzymatic reactions slow down at extremely high or low pH values?

 

 

 

 

 

 

  1. In what type of environment might an organism that produces this enzyme live? Explain your reasoning

Activity 4: Summary

 

 

 

 

 

 

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