Biomolecular condensates of Chlorocatechol 1,2-Dioxygenase as prototypes of enzymatic microreactors for the degradation of polycyclic aromatic hydrocarbons

Autor(es)

Evangelista, Nathan N. ; Micheletto, Mariana C. ; Kava, Emanuel ; Mendes, Luis F. S. ; Costa-Filho, Antonio J.

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 270, p. 9-pg., 2024-05-15.

Resumo

Polycyclic aromatic hydrocarbons (PAHs) are molecules with two or more fused aromatic rings that occur naturally in the environment due to incomplete combustion of organic substances. However, the increased demand for fossil fuels in recent years has increased anthropogenic activity, contributing to the environmental concentration of PAHs. The enzyme chlorocatechol 1,2-dioxygenase from Pseudomonas putida (Pp 1,2-CCD) is responsible for the breakdown of the aromatic ring of catechol, making it a potential player in bioremediation strategies. Pp 1,2-CCD can tolerate a broader range of substrates, including halogenated compounds, than other dioxygenases. Here, we report the construction of a chimera protein able to form biomolecular condensates with potential application in bioremediation. The chimera protein was built by conjugating Pp 1,2-CCD to low complex domains (LCDs) derived from the DEAD -box protein Dhh1. We showed that the chimera could undergo liquid -liquid phase separation (LLPS), forming a protein -rich liquid droplet under different conditions (variable protein and PEG8000 concentrations and pH values), in which the protein maintained its structure and main biophysical properties. The condensates were active against 4-chlorocatechol, showing that the chimera droplets preserved the enzymatic activity of the native protein. Therefore, it constitutes a prototype of a microreactor with potential use in bioremediation. (AU)

• Processo FAPESP: 17/24669-8 - Desvendando as bases moleculares de processos iniciais da secreção de proteínas em humanos utilizando técnicas biofísicas
• Beneficiário: Luis Felipe Santos Mendes
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 20/01152-2 - Construção e caracterização de micro-reatores da proteína clorocatecol 1,2-dioxigenase que utilizam domínios de baixa complexidade como adesivos moleculares
• Beneficiário: Nathan Nunes Evangelista
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 18/13016-6 - Ativação de Proteínas Fotossensíveis com Nanopartículas Radioluminescentes e Raios-X
• Beneficiário: Mariana Chaves Micheletto
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 15/50366-7 - Resolving mechanistic details of peptide transport across membranes using crystallographic and non-crystallographic structural biology approaches
• Beneficiário: Antonio José da Costa Filho
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 21/10795-7 - Lipidações e seus efeitos nas Proteínas de Estruturação e Compactação do Golgi (GRASPs)
• Beneficiário: Emanuel Kava
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 20/15542-7 - Construção e caracterização de micro-reatores da proteína clorocatecol 1,2-dioxigenase que utilizam domínios de baixa complexidade como adesivos moleculares
• Beneficiário: Antonio José da Costa Filho
• Modalidade de apoio: Auxílio à Pesquisa - Regular