Stepping into the Biocatalysis Lab for Undergraduate Students: From Enzyme Immobilization to Product Isolation
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[1] University of Pavia
University of Pavia
Pavía, Italia
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- Localización: Journal of chemical education, ISSN 0021-9584, Vol. 101, Nº 7, 2024, págs. 2790-2795
- Idioma: inglés
- Texto completo no disponible (Saber más ...)
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Resumen
A laboratory experiment was designed to introduce the practical application of biocatalysis in Bachelor and Master courses in biotechnology and medicinal chemistry, respectively. To this aim, the lipase from Candida rugosa was characterized (protein concentration and activity), immobilized on a carrier (by hydrophobic adsorption), and used in the regioselective hydrolysis of α-d-mannose pentaacetate into α-d-mannose 1,2,3,4-tetraacetate (∼60% yield), a valuable building block for the synthesis of oligosaccharides. This biotransformation occurs under mild conditions (room temperature, phosphate buffer containing 20% organic cosolvent, pH 4), requiring neither any protection/deprotection step nor specific equipment. The immobilized (heterogeneous) biocatalyst can be easily recovered from the reaction mixture by filtration and reused for further reaction cycles. Lab training was conceived to also include analytical and separation techniques routinely used in chemistry laboratories (chromatography, mass and NMR spectroscopy, titration, liquid–liquid extraction, filtration, and distillation under reduced pressure). At the end of the one-week lab training, students are expected to be able to carry out a simple enzymatic reaction (from the reaction setup to the product downstream) and to have acquired the practical basics of enzyme immobilization. Introducing students to biocatalysis aligns with current technological advancements, as biotransformations are being increasingly applied in the production of pharmaceuticals and commodity chemicals owing to the catalytic efficiency, selectivity, and ecocompatibility of enzymes.
