Collagenase Activity of Bromelain Immobilized at Gold Nanoparticle Interfaces for Therapeutic Applications

Autor(es)

Brito, Adrianne M. M. ^[1] ; Oliveira, Vitor ^[2] ; Icimoto, Marcelo Y. ^[2] ; Nantes-Cardoso, Iseli L. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed ABC, Ctr Ciencias Nat & Humanas, BR-09210580 Santo Andre, SP - Brazil

^[2] Univ Fed Sao Paulo, Escola Paulista Med, Dept Biofis, BR-04039032 Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PHARMACEUTICS; v. 13, n. 8 AUG 2021.
• Citações Web of Science: 0

Resumo

Bromelain (Bro) is a multiprotein complex extracted from the pineapple plant Ananas comosus, composed of at least eight cysteine proteases. Bro has a wide range of applications in medicine and industry, where the stability of its active proteases is always a major concern. The present study describes the improvement of stability and gain of specific activity in the enzymatic content of Bro immobilized on gold nanoparticles (GNPs). GNPs were synthesized in situ using Bro as the reducing and stabilizing agents and characterized by surface plasmon resonance and transmission electron microscopy. Consistent with the structural changes observed by circular dichroism analysis, the association with GNPs affected enzyme activity. The active Bro immobilized on GNPs (NanoBro) remained stable under storage and gained thermal stability consistent with a thermophilic enzyme. Proteolytic assays were performed on type I collagen membranes using fluorescence spectroscopy of O-phthaldialdehyde (OPA), changes in the membrane superficial structure, and topography by scanning electron microscopy, FTIR, and scanning laser confocal microscopy. Another characteristic of the NanoBro observed was the significant increase in susceptibility to the inhibitory effect of E-64, indicating a gain in cysteine protease activity. The higher stability and specific activity of NanoBro contributed to the broadening and improvement of Bro applications. (AU)

• Processo FAPESP: 18/09158-0 - Prolil oligopeptidase: estrutura-atividade e secreção
• Beneficiário: Vitor Marcelo Silveira Bueno Brandão de Oliveira
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 19/01487-7 - Nanopartículas de ouro associadas à cisteíno-proteases derivadas de Bromelaína: caracterização estrutural e ação colagenolítica
• Beneficiário: Marcelo Yudi Icimoto
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 19/14619-9 - Nanopartículas de ouro associadas à cisteíno-proteases derivadas de Bromelaína: caracterização estrutural e ação colagenolítica
• Beneficiário: Adrianne Marlise Mendes Brito
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico