Stairway to Stereoisomers: Engineering Short‐ and Medium‐Chain Ketoreductases To Produce Chiral Alcohols

Abstract

The short-and medium-chain dehydrogenase/reductase superfamilies are responsible for most chiral alcohol production in laboratories and industries. In nature, they participate in diverse roles such as detoxification, housekeeping, secondary metabolite production, and catalysis of several chemicals with commercial and environmental significance. As a result, they are used in industries to create biopolymers, active pharmaceutical intermediates (APIs), and are also used as components of modular enzymes like polyketide synthases for fabricating bioactive molecules. Consequently, random, semi-rational and rational engineering have helped transform these enzymes into product-oriented efficient catalysts. The rise of newer synthetic chemicals and their enantiopure counterparts has proved challenging, and engineering them has been the subject of numerous studies. However, they are frequently limited to the synthesis of a single chiral alcohol. The study attempts to defragment and describe hotspots of engineering short-and medium-chain dehydrogenases/reductases for the production of chiral synthons.

Microbial alcohol dehydrogenases: recent developments and applications in asymmetric synthesis

“…16 Shanbhag thoroughly described the engineering of short- and medium-chain dehydrogenases/reductases and their application in the production of chiral synthons. 17 Recently, Qiao et al reviewed protein engineering and the immobilization of keto reductases in the synthesis of chiral alcohols. 105 Here we shine a spotlight on the past five years of the engineering of microbial dehydrogenases towards the synthesis of chiral precursors of drugs and bioactive molecules with industrial applications.…”

“…Some recent reviews have updated the application of ketoreductases in asymmetric synthesis. [15][16][17]35,105 Li et al described the use of ketoreductases including engineered enzymes that were reported between 2018 and 2020, in the asymmetric synthesis of pharmaceuticals. 15 Another recent review elaborated on the use of wild type and engineered anti-Prelog ADHs in the synthesis of optically active alcohols.…”

“…Numerous short-chain ADHs or CRs have been engineered to produce evolved variants tailored for industrial applications. [14][15][16][17] Integrating computation knowledge, protein crystallography and an understanding of molecular insights into catalytic mechanisms have helped to generate enzyme variants with improved catalytic efficiency, stereoselectivity, substrate specificity and stability in the production of a significant number of chiral active pharmaceutical ingredients (APIs). Many interesting examples are presently available, which will be discussed in the light of scale-up possibilities in the manufacture of molecules of commercial importance.…”

Two anti‐Prelog NAD‐Dependent Alcohol Dehydrogenases with Broad Substrate Scope and Excellent Enantioselectivity

“…As mentioned above, another important distinction between ADHs is the preference for the coenzyme that can be NADH or NADPH. In general, most of the known natural ADHs are classified as Prelog and comprise both NAD‐ and NADP‐dependent enzymes [8c–e] . In contrast, anti‐Prelog ADHs are far less ubiquitous in nature and almost all of them are NADP(H) dependent [8d,10] .…”

The Molecular Basis of Catalysis by SDR Family Members Ketoacyl‐ACP Reductase FabG and Enoyl‐ACP Reductase FabI in Type‐II Fatty Acid Biosynthesis

“…Therefore, they are widely applied in the pharmaceutical industry as key chemical catalysts. [3,4] So far, only two SDR superfamily members have been confirmed to be involved in the molecular machineries for the analogous biosynthesis of fatty acids (FAS), polyketides (PKS) and non-ribosomal peptides (NRPS): the 3-ketoacyl reductase (KR) and enoyl reductase (ER). They are indispensable and conserved core enzymatic scaffolds in these metabolic routes for controlling energy supply, transcription and signaling.…”