Preparative Asymmetric Synthesis of 4,4-Dimethoxytetrahydro-2H-pyran-3-ol with a Ketone Reductase and in Situ Cofactor Recycling using Glucose Dehydrogenase

Abstract

The asymmetric enzymatic ketone reduction of 4,4-dimethoxytetrahydro-2H-pyran-3-one provided the (R)-r-hydroxyketal, an important chiral precursor for a pharmaceutical intermediate, with high enantioselectivity (>99% ee). An economical process including in situ NADPH-cofactor regeneration using glucose dehydrogenase has been developed to produce the desired material in high yield (96-98%). The two-enzyme process was employed at pilotplant scale to produce 80 kg of (R)-4,4-dimethoxytetrahydro-2Hpyran-3-ol. Critical factors for scale-up were found to be pH control and agitation speed.

Identification of ketone reductase ChKRED20 from the genome of Chryseobacterium sp. CA49 for highly efficient anti-Prelog reduction of 3,5-bis(trifluoromethyl)acetophenone

“…Requirements [2] Table S4) [7][8][9][10][11][12]. In addition, ChKRED20 could use 2-propanol as a cheap co-substrate for cofactor regeneration, and the crude enzyme could easily tolerate 40% (v/v) of 2-propanol in the reaction system.…”

“…In the last decade, successful application of ketone reductases (KRED) either as whole-cells or isolated enzymes has risen rapidly. In particular, several processes using isolated/crude recombinant enzymes coupled with efficient recycling systems have been developed into industrial-scale production with excellent stereoselectivity and economic advantage [7][8][9][10][11][12]. The use of recombinant enzymes greatly facilitates the scale-up with high volumetric productivity and the absence of side reactions [2,13,14].…”

Bioreductive production of enantiopure (S)-duloxetine intermediates catalyzed with ketoreductase ChKRED15

“…The bioreduction serves as a powerful approach for the production of optically active alcohols, which are widely used as fundamental intermediates for pharmaceuticals, agrochemicals and other products with chiral centers [1][2][3][4][5][6]. With the increased availability of recombinant ketoreductases, industrial-scale processes using cell-free enzymes coupled with efficient recycling systems have been developed rapidly in the past decade, which provides high volumetric productivity, excellent stereoselectivity, absence of side reactions and other economic advantages [7][8][9][10][11][12].…”

Rapid asymmetric reduction of ethyl 4-chloro-3-oxobutanoate using a thermostabilized mutant of ketoreductase ChKRED20

“…In addition, when significant improvement of catalytic properties is demanded, the recombinant enzymes can always be engineered via directed evolution to meet the process requirements. Hence, the application spectrum of the KREDs has been greatly expanded (Gooding et al 2010;Huisman et al 2010;Kosjek et al 2008;Liang et al 2010;Ma et al 2010;Solano et al 2012).…”