Lab Report I Guideline for
Isolation of Lysozyme from Hen Egg White.
Note: The lab report is to be computer generated and must be double-spaced.
Your report must include:
a) Title page
b) Introduction: Brief introduction explaining the purpose of the experiment and the
methodology of the techniques used. (Approximately 1 paragraph
c) Materials and Methods – “The procedure was followed according to……reference
to the manual. Include specific changes (these can be in point form).
d) Results and Discussion - include tables, figures. Sample calculations must be
included. Be sure to include sources of error that occurred in your experiment or that
could have potentially affected your results.
e) Conclusion: Should be a final statement of your results.
Tables, Figures, Calculations, Questions and Areas for Discussion
1. Include the bioinformatics search results for lysozyme. Be sure to provide the
appropriate references and web addresses for your answers.
a) How many references and pages are there? (Give the number of references (hits)
when you do a PubMed Search and then give two complete references).
b) What are the other sources of the enzyme? (List at least three)
c) How has it been purified? (Give two methods)
d) Has it been expressed in other organisms such as E.coli? (What other organisms?)
e) What is the sequence of the gene coding for it?
f) What is the expected protein sequence? (given) Using the BLAST tool, compare
the amino acid sequences to lysozyme from another source other than hen egg
white.
g) What is the metal ion present in the native enzyme? Is there a metal ion?
h) Find and cite correctly two references that study the inhibition of the enzyme.
i) What inhibitor molecules of the enzyme have been investigated?
j) What are other substrates for this enzyme?
k) What is the expected extinction coefficient for this enzyme in the next
experiment? (BioRad Protein Determination)
l) What is its expected molecular mass, PI, pH.
m) Has it been crystallized? How can it be crystallized? What does its crystal
structure look like?
2. Include Tables for enzyme reaction rates and protein concentrations
3. Standard Curve for Protein Determination
4. Prepare a Purification Table as outlined in the lab manual for each of the aliquots
based on the volumes and results from the enzyme activity and protein determination
assays. Compare activity results to "Standard Lysozyme".
5. Provide a drawing of the crystals you have grown, if you have been so lucky.
You may find purification tables and procedures on the net but if you are having
difficulty I will go through them next week.
If you have any questions, just ask, I will be happy to help.
Michelle