DEVELOPMENT OF A MACROPOROUS ADSORBENT AND CHEMICAL MODIFICATION WITH GLUTAMATE FOR PROTEIN PURIFICATION

The demand for biocomposites with high purity is increasing in the various industrial segments. Thus, chromatographic columns such as cryogenics have been developed. These have the advantage if modified chemically, seeking a breakthrough in purification processes. One example is cationic cryogel, obtained by immobilizing acid groups on its surface. To explore the capacity of the column, an alternative is the use for purification of lysozyme, an enzyme with a high isoelectric point, from its amino groups. Thus, the work aimed to develop a macroporous adsorbent functionalized with glutamate to purify the lysozyme of egg white. The cryogel was synthesized in a ratio of 7 % monomers with Acrylamide, N, N'-methylisocrilamide, and Alil glycidyl ether, and added ammonium persulfate and N, N, N', N' tetramethylene diamine. The solution was poured into a 10 mL syringe and placed in a thermostatic bath at -12 °C for 24 h. Subsequently, they were dried in an oven at 60 °C. The adsorbents were modified with ethylenediamine, glutaraldehyde, and glutamate and dried again. The egg white extract was obtained by removing the yolk and precipitation of the ovomucin. The efficiency of the process was evaluated by calculating the specific activity using Micrococcus lysodeikticus as substrate and by determining the purification factor. The crude extract showed a specific activity of 269.90 ± 2.858 U.mg-1, and after the purification process, a value of 2574.94 ± 11.425 U.mg-1 was obtained, and the result of the purification factor was 9.53. Therefore, the column presented satisfactory results regarding lysozyme purification, with an increase of almost ten times in its specific activity, indicating that the interaction mechanism by cation exchange was efficient for the process. Finally, the cryogenics produced have great potential for application in purification processes through cation exchange chromatography.