FTIR-ATR characterization of free Rhizomucor meihei lipase (RML), Lipozyme RM IM and chitosan-immobilized RML

The synthesis and characterization of biocatalysts based on lipase from Rhizomucor miehei (RML) immobilized on chitosan-based supports were investigated. The enzyme was immobilized on chitosan following two strategies: (i) physical adsorption; and (ii) covalent bonding using glutaraldehyde. The content of enzyme bound in the supports, as precipitable protein, was analyzed using UV/visible methods. FTIR-ATR spectroscopy was employed to characterize the prepared biocatalysts, as well the native enzyme and a commercial biocatalyst Lipozyme RM IM, used as reference materials. Analysis of the amide I′ signal allowed to follow the changes in the secondary structure of the enzyme after binding to the support and its thermal stability. The hydrolysis of ethyl stearate monitored in situ by FTIR-ATR was used as a test reaction. Results showed that RML was bound to Chit and Glut–Chit with minor changes in its secondary structure, thermal stability and enzymatic activity in a selected reaction test.

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• Rhizomucor miehei (RML) immobilized on chitosan-based supports.
• Secondary structure analysis was performed by FTIR-ATR spectroscopy.
• RML binds to the supports with minor changes in its secondary structure and thermal stability.
• The enzymatic activity for a hydrolysis test reaction was preserved.