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In vitro characterization of an enzymatic redox cascade composed of an alcohol dehydrogenase, an enoate reductases and a Baeyer-Villiger monooxygenase.

Oberleitner N, Peters C, Rudroff F, Bornscheuer UT, Mihovilovic MD - J. Biotechnol. (2014)

Bottom Line: An artificial enzyme cascade composed of an alcohol dehydrogenase, an enoate reductase and a Baeyer-Villiger monooxygenase was investigated in vitro to gain deeper mechanistic insights and understand the assets and drawbacks of this multi-step biocatalysis.Several substrates composed of different structural motifs were examined and provided access to functionalized chiral compounds in high yields (up to >99%) and optical purities (up to >99%).Hence, the applicability of the presented enzymatic cascade was exploited for the synthesis of biorenewable polyesters.

View Article: PubMed Central - PubMed

Affiliation: Institute of Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163-OC, 1060 Vienna, Austria.

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Scheme of the interplay of enzymes, substrates and cofactors.
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fig0005: Scheme of the interplay of enzymes, substrates and cofactors.

Mentions: As previously reported we implemented an in vivo enzymatic toolbox for redox cascade reactions (Oberleitner et al., 2013) composed of three enzymes. Within this study we demonstrated the flexibility and applicability of the cascade in asymmetric synthesis. In particular we investigated a three step cascade (Scheme 1) whereas the first reaction from an unsaturated cyclic alcohol to the corresponding ketone is catalyzed by two different alcohol dehydrogenases either LK-ADH from Lactobacillus kefir (Weckbecker and Hummel, 2006) or RR-ADH from Rhodococcus ruber (Stampfer et al., 2003). For the subsequent reduction of the unsaturated double bond we chose the well-known OYE1 from Saccharomyces carlsbergensis (Padhi et al., 2009) or XenB from Pseudomonas sp. (Oberleitner et al., 2013). For the last step, a Baeyer–Villiger oxidation, we employed CHMO from Acinetobacter sp. (Donoghue et al., 1976).


In vitro characterization of an enzymatic redox cascade composed of an alcohol dehydrogenase, an enoate reductases and a Baeyer-Villiger monooxygenase.

Oberleitner N, Peters C, Rudroff F, Bornscheuer UT, Mihovilovic MD - J. Biotechnol. (2014)

Scheme of the interplay of enzymes, substrates and cofactors.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263279&req=5

fig0005: Scheme of the interplay of enzymes, substrates and cofactors.
Mentions: As previously reported we implemented an in vivo enzymatic toolbox for redox cascade reactions (Oberleitner et al., 2013) composed of three enzymes. Within this study we demonstrated the flexibility and applicability of the cascade in asymmetric synthesis. In particular we investigated a three step cascade (Scheme 1) whereas the first reaction from an unsaturated cyclic alcohol to the corresponding ketone is catalyzed by two different alcohol dehydrogenases either LK-ADH from Lactobacillus kefir (Weckbecker and Hummel, 2006) or RR-ADH from Rhodococcus ruber (Stampfer et al., 2003). For the subsequent reduction of the unsaturated double bond we chose the well-known OYE1 from Saccharomyces carlsbergensis (Padhi et al., 2009) or XenB from Pseudomonas sp. (Oberleitner et al., 2013). For the last step, a Baeyer–Villiger oxidation, we employed CHMO from Acinetobacter sp. (Donoghue et al., 1976).

Bottom Line: An artificial enzyme cascade composed of an alcohol dehydrogenase, an enoate reductase and a Baeyer-Villiger monooxygenase was investigated in vitro to gain deeper mechanistic insights and understand the assets and drawbacks of this multi-step biocatalysis.Several substrates composed of different structural motifs were examined and provided access to functionalized chiral compounds in high yields (up to >99%) and optical purities (up to >99%).Hence, the applicability of the presented enzymatic cascade was exploited for the synthesis of biorenewable polyesters.

View Article: PubMed Central - PubMed

Affiliation: Institute of Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163-OC, 1060 Vienna, Austria.

Show MeSH