Dual enzyme multi-layer bioreactors: analytical modeling and experimental studies.

Area of Science:

• Biomedical Engineering
• Biotechnology
• Enzyme Kinetics

Background:

• Enzymic reactors are crucial for biomedical applications like blood detoxification.
• The Mercapturic Acid Pathway enzymes, Glutathione-S-Transferase (GST) and gamma-Glutamyl Transpeptidase (gamma GT), are suitable for this purpose.
• Previous studies focused on single immobilized GST reactors.

Purpose of the Study:

• To develop and analyze a dual-enzyme continuous packed-bed reactor (DCP) using GST and gamma GT in alternating layers.
• To investigate the effects of reactor configuration, kinetic parameters, and operational conditions on DCP performance.
• To validate theoretical models with experimental data.

Main Methods:

• Analytical modeling extended to a dual-enzyme system.
• Numerical simulations to study reactor configuration, kinetics (Km, Vmax, Kiq), and flow parameters.
• Experimental studies using immobilized GST (E1) and gamma GT (E2) on porous beads.
• Operation of the reactor in various configurations.

Main Results:

• Substrate and product concentration profiles were determined along the reactor.
• Reactor configuration (number of enzyme layers) was identified as a key performance factor.
• Inhibition of GST (E1) by its reaction product significantly affected DCP system performance.

Conclusions:

• DCP reactors offer a potential strategy for blood detoxification using Mercapturic Acid Pathway enzymes.
• Optimizing reactor configuration and managing enzyme inhibition are critical for efficient DCP operation.
• The study provides a theoretical and experimental framework for designing advanced enzymic reactors.