Asymmetric reductions using novel ene-reductases from cyanobacteria

Yilei Fu, doctoral thesis Technische Universität München, 2013

The asymmetric reduction of alkenes is attractive for producing chiral molecules, as up to two stereogenic centers can be created. Ten novel ene-reductases from various cyanobacterial strains were identified, expressed in Escherichia coli and purified. These enzymes were characterized regarding their substrate spectra, cofactor specificities and stereoselectivities. Biphasic whole-cell batch biotransformations of (R)-carvone using ionic liquids or adsorbent resins were developed up to the L-scale achieving up to 99.8 % conversion and 99.4 % diastereomeric excess (2R,5R-dihydrocarvone).

Publications

• Castiglione K, Fu Y, Polte I, Leupold S, Meo A, Weuster-Botz D (2017): Asymmetric whole-cell bioreduction of (R)-carvone by recombinant Escherichia coli with in situ substrate supply and product removal. Biochem Eng J 117: 102-111.
• Fu Y, Castiglione K, Weuster-Botz D (2015): Ene-reductases from cyanobacteria for industrial biocatalysis. Industrial Biocatalysis (Ed. Grunwald P), Pan Stanford Publishing Pte. Ltd., 631-661.
• Fu Y, Castiglione K, Weuster-Botz D (2013): Comparative characterization of novel ene-reductases from cyanobacteria. Biotechnol Bioeng 110: 1293-1301.
• Fu Y, Hölsch K, Weuster-Botz D (2012): A novel ene-reductase from Synechococcus sp. PCC 7942 for the asymmetric reduction of alkenes. Proc Biochem 47: 1988–1997