Blog Post 4:Lysozyme

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Q: What is Lysozyme? 
A: An enzyme found in saliva and sweat and tears that destroys the cell walls of certain bacteria.

Now that I have created what could possibly be lysozyme, my next step would be purifying it. Meaning I have to now find a protocol that would help me purify lysozyme that was extracted really is pure.  

This is one possible protocol:
 
Materials and Methods:
A solution of egg white diluted to ¼ with 0.1 M phosphate buffer pH 7 and filtered though glass wool is used as the source for lysozyme (Laboratory Manual. 2007). The solution is put through size exclusion chromatography with G-50 Sephadex column (fractionation range of 1,500-30,000 da) to produce 24 test tubes of equal egg white fractionations of 0.75 mL (Laboratory Manual. 2007). Numerous assays are conducted with varying pH and micrococcus (substrate) concentration to determine the optimal conditions for the highest enzymatic activity of lysozyme. After the collected column fractions and prepared egg white solution were left in the lab for two weeks, the odd numbered test tubes were assayed for lysozyme activity and the even numbered test tubes were assayed for protein at optimal pH. Refer to York University Department of Biology Laboratory Manual Summer 2008, SC/Biol 2020 Cell Biology and Biochemistry Pages 54-57 for a more detailed procedure of the lab. Also refer to the attached flow sheets for a thorough step by step procedure for this lab.

https://www.ukessays.com/essays/biology/purification-and-assaying-of-lysozyme-enzymes-biology-essay.php
 

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