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Chemical glycosidation of trypsin with O‐carboxymethyl‐poly‐β‐cyclodextrin: catalytic and stability properties

“…On the other hand, the ability of cyclodextrin to form an inclusion complex with BAEE (stability constant of the cyclodextrin–BAEE complex=180 M −1 [1]) could also generate an increased concentration of substrate at the microenvironment of the neoglycoenzyme and then decrease its K m value towards BAEE. A similar effect on the affinity for BAEE was previously reported for trypsin modified with monoactivated cyclodextrin derivatives [1,13,14].…”

“…The development of new strategies for preparing neoglycoenzymes, enzymes artificially linked to sugar moieties, has been a relevant biotechnological research topic during the last few years [1–3]. This interest has been inspired by the better stability and functional properties showed by the naturally occurring glycoenzymes in comparison with their non‐glycosidated counterparts [4].…”

“…This interest has been inspired by the better stability and functional properties showed by the naturally occurring glycoenzymes in comparison with their non‐glycosidated counterparts [4]. Several ionic [5–7] and non‐ionic polysaccharides [8–10], as well as monosaccharides and oligosaccharides [11–13], have been successfully used for modifying enzymes. On the other hand, different chemical [3,8,11], enzymic [14,15] and chemoenzymic [16] procedures have been employed for synthesizing neoglycoenzymes.…”

“…This is why the design of new sugar derivatives for transforming enzymes receives considerable attention in enzyme technology. We reported previously the chemical and the enzymic conjugation of hydrolytic enzymes with monoactivated cyclodextrin derivatives, demonstrating that the improved stability shown by these neoglycoenzymes was mediated by the occurrence of supramolecular associations in these conjugates [1,3,12–15].…”

Thermal stabilization of trypsin by enzymic modification with β‐cyclodextrin derivatives

“…In this regard, the formation of this kind of inclusion complex between the attached CD moieties and the hydrophobic side chains of several amino acid residues located at the surface of trypsin could cross‐link the protein structure, increasing the conformation rigidity of the modified enzyme. Recently, we confirmed the occurrence of this type of supramolecular association in proteases that were chemically modified with β‐cyclodextrin monoaldehyde, and their influence on the increased enzyme thermostabilization was well demonstrated [34,35].…”