Designing Multifunctional Enzymes through Fusion

CD Biosynsis is a leading expert in the field of enzyme design and evolution, with a long and in-depth experience in enzyme directed evolution. Our professional research team aims to combine the catalytic structural domains of different enzymes through enzyme fusion design to alter enzyme functions and provide valuable insights for the rational design of enzymes with improved properties or novel functions.

Overview

Molecular modification and redesign of enzymes is an important way to solve the bottleneck of enzyme catalysis for industrial applications. Fusion enzyme technology based on fusion protein design is a research hotspot of molecular enzyme engineering, which has been gradually applied to the construction and control research of multifunctional enzymes and enzyme proximity effects, showing important theoretical and applied research value. Designing multifunctional enzymes by fusion means combining the catalytic structural domains of different enzymes to create a single chimeric enzyme with multiple functions. This approach improves the efficiency and specificity of enzyme reactions and has advantages in various industrial and biotechnological applications.

Fig. 1 Fusing an alcohol dehydrogenase with an AT enables a cascade reaction starting from an alcohol to produce a chiral amine. (Aalbers F.S., et al., 2019)

Our Services

Our EnzymoGenius™ platform provides enzyme fusion design services that enable researchers to better conduct enzyme engineering studies and applications.

• Identify Target Enzymes
• Choose enzymes with complementary or synergistic functions.
• Consider enzymes that operate in similar or compatible conditions.
• Analyze Enzyme Structures
• Examine the crystal structures or predicted structures of the chosen enzymes.
• Identify the catalytic domains and other functional regions.
• Design Fusion Protein
• Select suitable linker regions between the different enzyme domains.
• Consider the orientation and order of the fused domains.
• Ensure that the fusion does not disrupt the active sites or functional regions.
• Optimize Linkers
• Use flexible or rigid linkers depending on the spatial arrangement of the catalytic domains.
• Optimize linker length to maintain the activity of each domain.
• Choose Expression System
• Bacteria
• Yeast
• Mammalian cells
• Construction and Clone
• Synthesize or clone the gene encoding the fused enzyme.
• Verify the correctness of the construct through sequencing.
• Expression and Purification
• Express the fusion protein in the chosen system.
• Optimize expression conditions for maximum yield.
• Purify the enzyme using standard techniques.
• Validation and Evaluation
• Validate the multifunctionality of the engineered enzyme.
• Assess the kinetic parameters, substrate specificity, and stability of the fusion enzyme.

Applications of Enzyme Fusion Design

Synthesis of Drug Intermediates: Multifunctional enzyme fusion design can be used to synthesize drug intermediates and active compounds.

Biofuel Production: Multifunctional enzyme fusion design can promote enzymatic reactions in biofuel production.

Food Processing: Multifunctional enzyme fusion design can be used in food processing to improve the efficiency of certain reactions.

Environmental Remediation: In wastewater treatment, multifunctional enzyme fusion design can be used to degrade pollutants.

CD Biosynsis strives to provide comprehensive and reliable enzyme fusion design services for a variety of applications. Contact us now to learn more about our enzyme fusion design services and how they can benefit your research or development program. Our team is ready to assist you and provide the insights that you need to achieve your goals.

Reference

1. Aalbers, F.S.; Fraaije, MW. Enzyme Fusions in Biocatalysis: Coupling Reactions by Pairing Enzymes. Chembiochem. 2019, 20(1):20-28.