Design and application of a bi-functional redox biocatalyst through covalent co-immobilization of ene-reductase and glucose dehydrogenase.

Autor: Nagy F; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, Hungary., Gyujto I; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, Hungary., Tasnádi G; Austrian Centre of Industrial Biotechnology, Austria; Department of Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria., Barna B; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, Hungary., Balogh-Weiser D; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, Hungary., Faber K; Department of Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria., Poppe L; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, Hungary; Biocatalysis and Biotransformation Research Center, Faculty of Chemistry and Chemical Engineering Babes-Bolyai University of Cluj-Napoca, Arany János str. 11, 400028 Cluj-Napoca, Romania. Electronic address: poppe@mail.bme.hu., Hall M; Department of Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria; Field of Excellence BioHealth, University of Graz, Austria. Electronic address: melanie.hall@uni-graz.at.
Jazyk: angličtina
Zdroj: Journal of biotechnology [J Biotechnol] 2020 Nov 10; Vol. 323, pp. 246-253. Date of Electronic Publication: 2020 Sep 03.
DOI: 10.1016/j.jbiotec.2020.08.005
Abstrakt: An immobilized bi-functional redox biocatalyst was designed for the asymmetric reduction of alkenes by nicotinamide-dependent ene-reductases. The biocatalyst, which consists of co-immobilized ene-reductase and glucose dehydrogenase, was implemented in biotransformations in the presence of glucose as source of reducing equivalents and catalytic amounts of the cofactor. Enzyme co-immobilization employing glutaraldehyde activated Relizyme HA403/M as support material was performed directly from the crude cell-free extract obtained after protein overexpression in E. coli and cell lysis, avoiding enzyme purification steps. The resulting optimum catalyst showed excellent level of activity and stereoselectivity in asymmetric reduction reactions using either OYE3 from Saccharomyces cerevisiae or NCR from Zymomonas mobilis in the presence of organic cosolvents in up to 20 vol%. The bi-functional redox biocatalyst, which demonstrated remarkable reusability over several cycles, was applied in preparative-scale synthesis at 50 mM substrate concentration and provided access to three industrially relevant chiral compounds in high enantiopurity (ee up to 97 %) and in up to 42 % isolated yield. The present method highlights the potential of (co-)immobilization of ene-reductases, notorious for their poor scalability, and complements the few existing methods available for increasing productivity in asymmetric bioreduction reactions.
(Copyright © 2020 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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