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Autor(en): Kahlow, Ulrich
Schmid, Rolf D.
Pleiss, Jürgen
Titel: A model of the pressure dependence of the enantioselectivity of Candida rugosa lipase towards (±)-menthol
Erscheinungsdatum: 2001
Dokumentart: Preprint
Erschienen in: Protein science 10 (2001), S. 1942-1952. URL http://dx.doi.org/10.1110%2Fps.12301
URI: http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-26743
http://elib.uni-stuttgart.de/handle/11682/849
http://dx.doi.org/10.18419/opus-832
Zusammenfassung: Transesterification of (±)-menthol using propionic acid anhydride and Candida rugosa lipase was performed in chloroform and water at different pressures (1, 10, 50, and 100 bar) to study the pressure dependence of enantioselectivity E. As a result, E significantly decreased with increasing pressure from E=55 (1 bar) to E=47 (10 bar), E=37 (50 bar), and E=9 (100 bar). In order to rationalize the experimental findings, molecular dynamics simulations of Candida rugosa lipase were carried out. Analyzing the lipase geometry at 1, 10, 50, and 100 bar revealed a cavity in the Candida rugosa lipase. The cavity leads from a position on the surface distinct from the substrate binding site to the core towards the active site and is limited by F415 and the catalytic H449. In the crystal structure of the Candida rugosa lipase, this cavity is filled with 6 water molecules. The number of water molecules in this cavity gradually increased with increasing pressure: 6 molecules in the simulation at 1 bar, 10 molecules at 10 bar, 12 molecules at 50 bar, and 13 molecules at 100 bar. Likewise, the volume of the cavity progressively increased from about 1864 ų in the simulation at 1 bar to 2529 ų at 10 bar, 2526 ų at 50 bar, and 2617 ų at 100 bar. At 100 bar, one water molecule slipped between F415 and H449, displacing the catalytic histidine side chain and thus opening the cavity to form a continuous water channel. The rotation of the side chain leads to a decreased distance between the H449-N and the (+)-menthyl-oxygen (non-preferred enantiomer) in the acyl enzyme intermediate, a factor determining the enantioselectivity of the lipase. While the geometry of the preferred enantiomer is similar in all simulations, the geometry of the non-preferred enantiomer gets gradually more reactive. This observation correlates with the gradually decreasing enantioselectivity E.
Enthalten in den Sammlungen:03 Fakultät Chemie

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