The synthesis of a D-amino acid ester in an organic media with ?-chymotrypsin modified by a bio-imprinting procedure

Interfacial activation-based molecular bioimprinting of lipolytic enzymes.

“…In particular, one interesting facet of the exploitation of the drastically lowered flexibility of a protein in very low-water media is the so-called ligand-induced enzyme "memory" (or simply ligand memory) (6), presumably based on inducing, upon ligand binding, an enzyme conformational change that (after freeze-drying of the solution) will be preserved in the lyophilized sample when exposed to anhydrous solvents. This strategy, also referred to as molecular (bio)imprinting (7,8), has been extended to nonenzymic proteins and other macromolecules (9, 10) and shares a conceptual analogy with the previously known molecular imprinting of polymers (for a review, see ref. 11).…”

“…11). In the case of enzyme (bio)imprinting, only scarce efforts have been reported so far, which have focused on a very few related proteases and made use of competitive inhibitors (amino acid derivatives) as print molecules (6)(7)(8). Although these preliminary results seem indeed encouraging in terms of an activity enhancement or even an apparent (stereo)selectivity alteration in the organic milieu, the nature of the presumptive conformational changes induced has not been characterized, and the molecular reasons for the experimental observations are still open to debate (12).…”

Enhancing the catalytic efficiency of subtilisin for transesterification by dual bioimprinting

“…25,27,33,36 Transesterification in the case of alcoholysis involves two substrates-an ester and an alcohol which acts as a nucleophile. Hence, the nucleophile alcohol should also be able to bioimprint the enzyme.…”

Induction of catalytic activity in proteins by lyophilization in the presence of a transition state analogue

“…It can be best illustrated by (bio)molecular imprinting, an approach which enables the introduction of new binding sites into protein molecules (Braco et al, 1990;Dabulis and Klibanov, 1992) and the alteration of an enzyme's catalytic properties such as activity (Mingarro et al, 1995), substrate specificity (Johnsson et al, 1995;O'Rich, and Dordick, 1997) or enantioselectivity (Stahl et al, 1990(Stahl et al, , 1991. However, if ligands can be employed to alter protein conformations in organic solvents in a predictable manner, it should be possible to use this approach for the induction of ''de novo'' catalytic activity in existing proteins via imprinting with transition state analogues (TSA) (Lerner et al, 1991).…”