Isolation and Purification of the Lysozyme

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Enzyme purification Lab:

Isolation and purification of the lysozyme


This laboratory included the ways in purifying and assaying enzymes using egg white. Lysozyme cleaves Micrococcus lysodeikticus by destroying their cell walls, which essentially destroys them. Gel chromatography was used to elute different proteins by mass. The results show that the highest enzyme activity was on fraction number. They also show that the highest protein concentration was at 8. It is possible that each fraction would contain other proteins in addition to lysozyme.


The purpose of this lab is to examine different methods of purifying enzyme and from that purified samples investigate the lysozyme activity and protein concentration [8]. Different methods have been used in the past to purify the egg white such as gel chromatography, ion exchange chromatography, and Affinity chromatography [8]. The chromatography that was used during this experiment to separate the proteins of the egg white is known as the Sephadex G-50 column chromatography. In this chromatography, the particles are being separated according to their size. The bigger the protein, faster it will elute. The gel in the sephadex column has micro pores. The small proteins go in and out of these pores and thus elute later. Egg white contains different protein in different concentration. It contains 54% ovalbmin, 13% conalbumin, 11% ovomucoid, 8% globulins and 3.5% lysozyme [4].

The protein of interest in this lab is lysozyme.Lysozyme was abstracted from egg white solution using the Sephadex G-50 column chromatography [8]. It can be seen from the concentration of proteins that the least amount of lysozyme(3.5%) is found in the egg white.[4] The lysozyme has the smallest molecular weight compared to other proteins found in egg white. It contains 129 amino acids [2]. Since lysozyme is an enzyme it has a three dimensional...