Cytochrome P450 Monooxygenases Catalyse Steroid Nucleus Hydroxylation with Regio‐ and Stereo‐Selectivity

Abstract

Steroids are the second largest class of drugs with a wide range of pharmacological properties. Hydroxylation of steroids seriously affects their biological activities and other properties. However, steroids are mostly sp 3 hybridized carbons with numerous CÀ H bonds far from the functional group that can activate them, and achieving regio-and stereo-selective hydroxylation on steroids is a highly challenging task that is almost impossible to achieve using modern organic synthesis techniques. Interestingly, cytochrome P450 monooxygenases possess the ability to catalyse regio-and stereo-selective oxidations of nonactivated CÀ H bonds in complex organic molecules under mild conditions. This review summarizes the P450s identified and engineered in recent years that can catalyse steroid nucleus hydroxylation stereo-and regio-selectively.1.

Chemoenzymatic Synthesis of Steroidal Products: Recent Advances

“…With the advancements in genomics and transcriptomics, an increasing number of steroid‐modifying enzymes have been discovered. These include the side chain degrading enzymes (P450 SCC ) found in plants and animals, aldoase (Atf1) from Mycobacteria , 3‐Ketosteroid‐Δ1‐dehydrogenases (KstD) found in Mycobacteria and Propionibacterium , and various steroidal hydroxylases, promoting the diversified production of important steroid molecules [9c,32] . It is worth mentioning that multiple steroid‐hydroxylase mutants with new functions were created based on enzyme engineering for the efficiently synthesis of target steroids [7,33] .…”

Choose Your Own Adventure: A Comprehensive Database of Reactions Catalyzed by Cytochrome P450 BM3 Variants

“…Given the attractive features of P450 BM3, the enzyme has garnered significant attention and has been the subject of numerous reviews highlighting its exceptional properties and characteristics. Reviews have been conducted on various aspects of P450 BM3, including structural and mechanistic details, − hydroxylation, ,,− pharmaceutical compound hydroxylation, ,− epoxidation, ,,, and carbene/nitrene transfer. ,,,,,,− Additionally, studies have explored the use of decoy molecules to improve enzymatic activity, ,,,,,,− as well as P450 BM3-inorganic complexes. − Reviews of P450 BM3 frequently place it within a broader context of other P450 enzymes, resulting in a lack of focus. Though some reviews do describe reactions catalyzed by P450 BM3 variants, they are typically limited to discussion of the “best” variants for a given reaction and do not offer a comprehensive review of reported variants.…”

Recent developments in the enzymatic modifications of steroid scaffolds

“…Hydroxylation of steroids stands as one of the most pivotal reactions in modifications of the steroid scaffold. [15][16][17] The addition of hydroxy groups not only influences physiological functions, polarity, solubility, and toxicity but also serves as a synthetic starting point for the preparation of valuable steroid drugs. 18,19 In this section, we discuss recent advances in the discovery and engineering of cytochrome P450 oxygenases (P450s), non-heme iron-and α-ketoglutarate-dependent oxygenases (αKG OXs), Rieske oxygenases, and other enzymes for site-specific hydroxylations.…”

“…18,19 In this section, we discuss recent advances in the discovery and engineering of cytochrome P450 oxygenases (P450s), non-heme iron- and α-ketoglutarate-dependent oxygenases (αKG OXs), Rieske oxygenases, and other enzymes for site-specific hydroxylations. For a comparative summary elucidating the preference sites for enzymatic hydroxylation of steroids, readers are directed to the 2020 review by Zhang et al , the 2022 review by Zhu et al and the 2023 review by Abas et al 15–17…”