A novel lipase-based stationary phase in liquid chromatography

Area of Science:

• Biocatalysis and Separation Science
• Enzyme Immobilization Technology

Background:

• Chiral compounds require specialized separation techniques due to their distinct biological activities.
• Enzyme immobilization offers a stable and reusable platform for biocatalytic applications.
• Developing efficient chiral stationary phases (CSPs) is crucial for enantioseparation.

Purpose of the Study:

• To develop a novel chiral stationary phase (CSP) based on immobilized Candida antarctica lipase B (CALB).
• To evaluate the stereoselectivity of the developed CSP in enantioseparation and asymmetric hydrolysis.
• To assess the potential of the CSP as a biocatalytic reactor.

Main Methods:

• Immobilization of Candida antarctica lipase B (CALB) onto macroporous silica gel using the "in batch" method.
• Preparation of three CALB-based High-Performance Liquid Chromatography (HPLC) columns with varying enzyme loadings.
• Utilizing epoxy silica and aminopropyl silica as chromatographic supports.
• Evaluating enantioseparation of aromatic alcohols and diniconazole enantiomers.
• Assessing asymmetric hydrolysis of chiral esters.

Main Results:

• Successful development of a novel CSP by immobilizing CALB on macroporous silica gel.
• Demonstrated stereoselectivity of the CALB-based CSP in separating aromatic alcohols and diniconazole enantiomers.
• Confirmed the capability of the CSP to perform asymmetric hydrolysis of chiral esters, functioning as a bioreactor.
• The "in batch" immobilization method proved effective for enzyme attachment to both epoxy and aminopropyl silica supports.

Conclusions:

• The developed CALB-based CSP is effective for both enantioseparation and asymmetric hydrolysis.
• The CSP shows significant potential as a reusable and efficient biocatalytic tool.
• This approach offers a promising strategy for chiral analysis and synthesis.