Teixeira Polez_2025_Macromol.Biosci__e00221

This study utilizes cellulose sourced from cotton linters to synthesize cellulose esters-hexanoate, benzoate, and mixed hexanoate-benzoate-with varying degrees of substitution (DS). These esters create electrospun mats that immobilize Pseudomonas fluorescens lipase (PFL), also in a configuration where an intermediate layer is added to a mat using an airbrush filled with PFL, covered by a third layer of electrospun mat. PFL-incorporated spheres are produced from cellulose ester solutions. DS, acyl chain length, and electrospinning parameters influence the morphology of the electrospun mat, which consists of nanofibers and ultrafine fibers. The PFL-incorporated mats show poor catalytic activity in resolving racemic (R,S)-2-chloro-1-phenylethanol, likely due to enzyme deactivation from high-voltage electrospinning. In contrast, mat-layered structures with PFL immobilized without voltage nearly doubled the conversion rate, although it was still lower than that of free enzymes. Spheres enhanced biocatalysis, achieving a 40% conversion rate with 94% enantiomeric purity while retaining 76% of their initial conversion rate in a subsequent reaction cycle. This research is the first to explore cellulose esters for the enzymatic immobilization of PFL to resolve a racemic mixture. The findings may enable PFL-incorporated structures in broader biocatalysis applications; the materials created may be tested to support the immobilization of other enzymes.