Original scientific paper

The Use of Lewis Cell to Investigate the Enzyme Kinetics of an (S)-Hydroxynitrile Lyase in Two-Phase Systems

Michael Bauer ; Institute of Biotechnology, Enzyme-Technology Group, SFB Biocatalysis Graz University of Technology, Petersgasse 12, A-8010 Graz, Austria
Roland Geyer ; Institute of Biotechnology, Enzyme-Technology Group, SFB Biocatalysis Graz University of Technology, Petersgasse 12, A-8010 Graz, Austria
Herfried Griengl ; Institute of Organic Chemistry, SFB Biocatalysis Graz University of Technology, Stremayrgasse 16, A-8010 Graz, Austria
Walter Steiner ; Institute of Biotechnology, Enzyme-Technology Group, SFB Biocatalysis Graz University of Technology, Petersgasse 12, A-8010 Graz, Austria

Abstract

(S)-Hydroxynitrile lyase from Hevea brasiliensis (HbHnl) (EC 4.1.2.39) catalyzes the reversible synthesis of chiral cyanohydrins from aldehydes or ketones and HCN. The enzymatic formation of (S)-mandelonitrile (MN) from benzaldehyde (BA) and HCN was studied in two-phase systems of buffer and organic solvents (diisopropyl ether, methyl-t-butyl ether) using a Lewis cell to investigate the interaction between mass transfer and the biocatalytic reaction. The enzymatic reaction rate in the aqueous phase saturated with organic solvents is drastically reduced in comparison to pure buffer due to the increase of the Michaelis-Menten constants of the substrates. Mass transfer of the substrates from the organic to the aqueous phase and mass transfer of the product in the opposite direction could be described by the two-film theory. The formation of (S)-mandelonitrile in the Lewis cell follows an aqueous phase distributed reaction model, which means that the enzymatic reaction takes place in the bulk of the aqueous phase and in the thin film close to the interface and/or directly at the interface. Using the Hatta number it could be shown that the mass transfer of benzaldehyde from the organic to the aqueous phase is enhanced by the biocatalytic reaction of the (S)-hydroxynitrile lyase from Hevea brasiliensis.

Keywords

Lewis cell; two-phase system; mass transfer; enzyme kinetics; hydroxynitrile lyase; Hevea brasiliensis

Hrčak ID:

178384

URI

https://hrcak.srce.hr/178384

Publication date:

14.3.2002.

Article data in other languages: croatian

Visits: 1.145 *