Magnetic Multi-Enzymatic System for Cladribine Manufacturing

Area of Science:

• Biocatalysis and Enzyme Engineering
• Sustainable Chemistry
• Pharmaceutical Synthesis

Background:

• Enzyme-mediated processes offer sustainable alternatives to traditional chemical synthesis.
• Multi-enzymatic systems enhance complex synthetic pathways by minimizing by-products and overcoming reaction limitations.
• Immobilizing enzymes on magnetic supports facilitates catalyst reusability and simplifies downstream processing.

Purpose of the Study:

• To develop a novel cascade system for cladribine synthesis using immobilized enzymes.
• To optimize magnetic biocatalysts for efficient and sustainable pharmaceutical production.
• To evaluate the reusability and efficiency of the developed enzyme system.

Main Methods:

• Immobilization of purine 2'-deoxyribosyltransferase (LmPDT) and hypoxanthine phosphoribosyltransferase (EcHPRT) onto magnetic microspheres.
• Biochemical characterization of magnetic biocatalyst derivatives (MLmPDT3 and MEcHPRT3).
• Application of the MLmPDT3/MEcHPRT3 system for one-pot cladribine synthesis.

Main Results:

• Optimized magnetic biocatalysts MLmPDT3 and MEcHPRT3 demonstrated high activity and stability.
• The MLmPDT3/MEcHPRT3 system achieved a 1.67-fold improvement in cladribine synthesis compared to MLmPDT3 alone.
• The system efficiently converted an undesired by-product into a high-added-value product (90% conversion of Hyp into IMP).
• The magnetic biocatalyst system retained 75% activity after 16 reuse cycles.

Conclusions:

• The developed magnetic biocatalyst cascade system provides an efficient and sustainable method for cladribine synthesis.
• Enzyme immobilization on magnetic supports enhances catalyst reusability and process efficiency.
• This approach offers a greener alternative for pharmaceutical manufacturing by minimizing waste and maximizing product value.