Dados do Trabalho

Título

SUPERMACROPORE BIOREACTORS: ENZYME IMMOBILIZATION WITH ANTIMICROBIAL POTENTIAL

Introdução

Growing industrialization has increased the demand for sophisticated biotechnological processes, thus different methods for using enzymes have emerged. The development of enzymatic bioreactors is an alternative that aims to reduce costs, since there are no losses to the reaction medium. To this end, the use of macroporous cryogels presents good results, making their use viable in enzyme immobilization processes. Lysozyme is known for its antimicrobial activity, which comes from the cleavage of the cell wall of certain bacteria, and can be applied in conservation methods. Therefore, the work aimed to develop bioreactors via lysozyme immobilization and evaluate its antimicrobial potential.

Material e Métodos

The cryogels were prepared in a proportion of 7% monomers with acrylamide, N, N'-methylisocrylamide, and Allyl 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 functionalizations followed the glutaraldehyde and IMAC method using IDA and iron. To produce the bioreactors, 1 mg/mL of pure lysozyme was kept in contact with the supports for 14 h and 25 °C, and subsequently washed. The efficiency of the process was verified by the concentration of enzyme retained on the support and verification of activity using Micrococcus lysodeikticus as substrate.

Resultados e Discussão

The bioreactors produced with glutaraldehyde and IMAC obtained an adsorptive capacity of 30.55±2.50 mg/g and 25.84±1.13 mg/g respectively, while the control was 3.07±0.08 mg/g, This demonstrates that the insertion of reactive groups on the surface of the support is responsible for greater enzymatic immobilization.

Conclusão

The bioreactors were subjected to 6 cycles of use, and it was observed that the free enzyme presented 116.4 ±0.86 U of enzymatic activity and that there was no statistical difference for the two bioreactors produced, that is, only one cycle of use was performed. sufficient to supply the efficiency of free lysozyme. Therefore, the continuous use of bioreactors proved to be beneficial and efficient in bacterial cell lysis, indicating great potential for use for antimicrobial purposes.