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Published on: February 11, 2019
Recombinant laccase rPOXA 1B real-time, accelerated and molecular dynamics stability study
Leidy D Ardila-Leal1, Pedro A Monterey-Gutiérrez2, Raúl A Poutou-Piñales3
1Departamento de Microbiología. Facultad de Ciencias. Pontificia Universidad Javeriana (PUJ). Bogotá, Laboratorio de Biotecnología Molecular, Grupo de Biotecnología Ambiental e Industrial (GBAI), Bogotá, D.C, Colombia.
The laccase enzyme rPOXA 1B shows remarkable stability across various temperatures, with half-life values up to 230.8 months. This stability is crucial for its industrial applications in biotechnology.
Area of Science:
- Biotechnology
- Enzymology
- Biochemistry
Background:
- Laccases (EC 1.10.3.2) are multi-copper oxidoreductases with significant biotechnological applications.
- Their high oxidative potential makes them valuable for degrading pollutants and synthesizing compounds.
Purpose of the Study:
- To assess the real-time stability (RTS) and accelerated stability (AS) of the laccase enzyme rPOXA 1B.
- To investigate the enzyme's stability under various temperature conditions using experimental and computational methods.
Main Methods:
- Real-time stability study conducted over one year at five different temperatures (243.15–313.15 K).
- Accelerated stability (AS) calculated using RTS data and the Arrhenius equation.
- Molecular dynamics (MD) simulations performed at four different temperatures (241–314 K).
Main Results:
- Laccase rPOXA 1B retained significant activity at tested temperatures, with recovery rates varying from 101.16% to 4.83%.
- AS study indicated high stability at 240.98 K (t½=230.8 months), 277.40 K (t½=46.2 months), and 297.53 K (t½=12.6 months).
- MD simulations correlated well with experimental data, showing increased residue fluctuations with rising temperatures.
Conclusions:
- Laccase rPOXA 1B is a highly stable enzyme, confirmed by both experimental and computational analyses.
- The enzyme's stability is suitable for preservation without preservatives at relevant temperatures, offering utility for large-scale producers.
