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

Author(s)

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

• Document type: Journal article
• Source: International Journal of Biological Macromolecules; v. 270, p. 9-pg., 2024-05-15.

Abstract

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)

• FAPESP's process: 17/24669-8 - Unraveling the molecular bases of the early protein secretory pathway in humans using biophysical techniques
• Grantee: Luis Felipe Santos Mendes
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 20/01152-2 - Construction and characterization of micro reactors of chlorocatechol 1,2-dioxygenase that utilize low complexity domains as molecular adhesives
• Grantee: Nathan Nunes Evangelista
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 18/13016-6 - Activation of Photosensitive Proteins with Radioluminescent Nanoparticles and X-Rays
• Grantee: Mariana Chaves Micheletto
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 15/50366-7 - Resolving mechanistic details of peptide transport across membranes using crystallographic and non-crystallographic structural biology approaches
• Grantee: Antonio José da Costa Filho
• Support Opportunities: Regular Research Grants
• FAPESP's process: 21/10795-7 - Lipidations and their effects on the Golgi Reassembly and Stacking Proteins (GRASPs)
• Grantee: Emanuel Kava
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 20/15542-7 - Design and characterization of micro-reactors constituted by the protein chlorocatechol 1,2-dioxygenase and low complexity domains
• Grantee: Antonio José da Costa Filho
• Support Opportunities: Regular Research Grants