Enabling Selective and Sustainable P450 Oxygenation Technology. Production of 4-Hydroxy-α-isophorone on Kilogram Scale

Abstract

The development of P450 platform technology has enabled the sustainable production of an oxygenated intermediate, 4-hydroxy-α-isophorone, on kilogram scale. Application of a cytochrome P450 enzyme resulted in an unprecedented product concentration of 10 g/L and a space–time yield of 1.5 g/L/h. These findings are highly relevant for the economical evaluation of cytochrome P450 technology and, additionally, provide access to an alternative and cost-effective route toward 4-hydroxy-α-isophorone.

Ketoisophorone Synthesis with an Immobilized Alcohol Dehydrogenase

“…[10][11][12] Apart from its biomass origin, αisophorone is also produced at large scale as a waste recovery operation from industry. [19,20] The biocatalyst utilized consisted of E. coli whole cells overexpressing a P450 BM3 and a Glucose dehydrogenase, that served as the cofactor regeneration enzyme. [13] Once the KET has been synthesized, it can then be reduced to (4R,6R)-actinol which is an intermediate for the production of zeaxanthin, cryptoxanthin and xanthoxin.…”

Catalytic Aerobic Oxidation of C(sp3)−H Bonds

“…[143] It should be noted however that while the enzymes may be recoverable,the cofactors are often not so,meaning they need to be added in each batch. [144] To avoid superstoichiometric addition of NADPH cofactor in this oxidation reaction, heterologous coexpression of aglucose dehydrogenase (GDH) was used to allow cofactor regeneration with glucose.T his process was limited by O 2 mass transfer and self-deactivation of the catalyst. [144] To avoid superstoichiometric addition of NADPH cofactor in this oxidation reaction, heterologous coexpression of aglucose dehydrogenase (GDH) was used to allow cofactor regeneration with glucose.T his process was limited by O 2 mass transfer and self-deactivation of the catalyst.…”

Whole‐cell biocatalysis platform for gram‐scale oxidative dearomatization of phenols

“…Examples of WC biocatalytic processes. (a) Select scaffolds accessed through WC biocatalytic reactions (Baker Dockrey et al., ; Guo et al., ; Hadi et al., ; Kaluzna et al., ; Matsuyama et al., ; Milker et al., ; Xie & Tang, ). (b) WC biocatalysis utilizing a multi‐enzyme cascade to generate chiral benzylic amines (Both et al., ) [Colour figure can be viewed at wileyonlinelibrary.com]…”

“…On an industrial scale, several processes have been designed to employ WC biocatalysts. Notably, WC biocatalysis has been utilized for the generation of valuable chemical feedstocks such as enantio‐enriched alcohols, (Matsuyama, Yamamoto, & Kobayashi, ) performance material monomers, (Kaluzna et al., ) and pharmaceutically relevant intermediates, (Guo et al., ) as well as the late‐stage functionalization of active pharmaceutical ingredients during route development (Figure a; Guo et al., ; Milker, Fink, Rudroff, & Mihovilovic, ; Xie & Tang, ). Recently, researchers at GlaxoSmithKline demonstrated that a WC biocatalysis platform is also useful for discovery of new synthetic routes on the gram‐scale, developing several biocatalytic routes to chiral 1,3‐substituted cyclohexanones, such as 2 (Hadi et al., ).…”