Homework SourseA Effect of Enzyme Concentration The effect of the enzyme co
A. Effect of Enzyme Concentration:
The effect of the enzyme concentration on the initial velocity of the reaction will be determined for the commercially prepared lysozyme. The concentration of the commercially available lysozyme will be varied from .06mg/ml to 1.8 mg/ml. Using the available stock solutions, prepare a procedure to test the effect of varying lysozyme concentration on its functional activity. Complete the protocol Table, Part A onpage 63.
Calculate the reagent and enzyme concentrations in the assay tubes and add these to the protocol. Use the calculation procedure described below. To calculate the enzyme concentrations, protein concentrations in the assay tubes will be calculated. First, calculate the amount of protein (mg) added to each assay tube. This is determined by taking the concentration of the stock enzyme solution (30 mg/ml for the commercially prepared lysozyme) and multiplying that value by the volume of stock enzyme used in that assay. To determine the protein concentration in the assay tube divide the number of mg of protein in the assay tube by the assay volume (5.0 ml). As you complete Part A Table, remember to account for two sources of NaCl.
Key Concept Learning Outcomes:
Calculation of [NaCl] and [buffer] from two sources
If the concentrations are identical, then the volumes can be added.
Ex: .10 ml of lysozyme in .1M NaCl and .90 ml of .1M NaCl= 1.0 ml .1M NaCl
If the concentrations are different, calculate moles from each source
I NEED HELP FINISHING THE TABLE AND SAMPLE CALCULATIONS
PART A. EFFECT OF ENZYME CONCENTRATION ([S] constant)
Reagent Test tube #
1 2 3 4 5 6
.10 M NaCl
(lysozyme in .10M NaCl)
2.99ml
2.98
2.95
2.90
2.80
2.70
[NaCl] (total)
.50 mg/ml cell wall substrate
in .15 M sodium phosphate,pH 6.2
2.00
2.00
2.00
2.00
2.00
2.00
[cell wall substrate]
[sodium phosphate]
Lysozyme (30 mg/ml)
In .10M NaCl
.01
(10 mL)
.02
.05
.10
.20
.30
[lysozyme]
(mg/ml)
| Reagent Test tube # | ||||||
| 1 2 3 4 5 6 | ||||||
| .10 M NaCl (lysozyme in .10M NaCl) | 2.99ml | 2.98 | 2.95 | 2.90 | 2.80 | 2.70 |
| [NaCl] (total) | ||||||
| .50 mg/ml cell wall substrate in .15 M sodium phosphate,pH 6.2 | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 |
| [cell wall substrate] | ||||||
| [sodium phosphate] | ||||||
| Lysozyme (30 mg/ml) In .10M NaCl | .01 (10 mL) | .02 | .05 | .10 | .20 | .30 |
| [lysozyme] (mg/ml) |
Solution
The problem is not clear. However solution for final enzyme concentration is given here
30mg/ml----if we take 0.01 ml, it will contain 0.3 mg of lysozyme.this is in 10 ml of 0.1M Nacl. For 1ml-----0.3mg/10=0.03mg lysozyme/ml of 0.1MNacl. results are in table
Lysozyme (30 mg/ml)
In .10M NaCl
.01
(10 mL)
.02
.05
.10
.20
.30
[lysozyme]
(mg/ml)
0.03mg/ml
0.06mg/ml
0.15mg/ml
0.3mg/ml
0.6mg/ml
0.9mg/ml
| Lysozyme (30 mg/ml) In .10M NaCl | .01 (10 mL) | .02 | .05 | .10 | .20 | .30 |
| [lysozyme] (mg/ml) | 0.03mg/ml | 0.06mg/ml | 0.15mg/ml | 0.3mg/ml | 0.6mg/ml | 0.9mg/ml |
