Industrial Application of 2-Oxoglutarate-Dependent Oxygenases

Abstract

C–H functionalization is a chemically challenging but highly desirable transformation. 2-oxoglutarate-dependent oxygenases (2OGXs) are remarkably versatile biocatalysts for the activation of C–H bonds. In nature, they have been shown to accept both small and large molecules carrying out a plethora of reactions, including hydroxylations, demethylations, ring formations, rearrangements, desaturations, and halogenations, making them promising candidates for industrial manufacture. In this review, we describe the current status of 2OGX use in biocatalytic applications concentrating on 2OGX-catalyzed oxyfunctionalization of amino acids and synthesis of antibiotics. Looking forward, continued bioinformatic sourcing will help identify additional, practical useful members of this intriguing enzyme family, while enzyme engineering will pave the way to enhance 2OGX reactivity for non-native substrates.

Structural basis for divergent C–H hydroxylation selectivity in two Rieske oxygenases

“…nzymes that perform C-H hydroxylation reactions often demonstrate impressive control over site-and stereoselectivity on complex scaffolds [1][2][3] . This precision is emulated in numerous natural product biosynthetic pathways by several classes of metalloenzymes including cytochrome P450 monooxygenases 4 , non-heme α-ketoglutarate-dependent oxygenases 5 , and Rieske oxygenases 6 .…”

“…1), displays divergent selectivity. GxtA hydroxylates the C11 β-position of β-STOH (1) and STX (2) to afford 11-β-hydroxy-β-saxitoxinol (3) and 11-β- (Pfam PF00355) generated by the EFI-EST 59 server at an E-value of 5 and alignment score of 65, visualized in Cytoscape v3.7.1 60 . The network shown was trimmed to include only Rieske proteins that contain the non-heme iron catalytic domain (IPR017941, IPR005805, and IPR domains listed in the figure).…”

Overview on Multienzymatic Cascades for the Production of Non-canonical α-Amino Acids

“…Enzymes from biosynthetic pathways to NcAAs ( Hedges and Ryan, 2020 ) are clear candidates for MECs expansion. On the other hand, information of specific enzymes for functionalization of AAs or AA transformation into other important pharmaceutical compounds is also accumulating in the literature (e.g., Smirnov et al, 2012 ; Busto et al, 2014 ; Hönig et al, 2017 ; Hyslop et al, 2018 ; Parthasarathy et al, 2018 ; Peters and Buller, 2019 ; McDonald et al, 2019 ; Hedges and Ryan, 2020 ; Mindt et al, 2020 ; Song et al, 2020 ; Wendisch, 2020 ; Zhang et al, 2020 ; Zhao et al, 2020 ). These enzymes are, in principle, complementary to well stablished NcAA-production MECs.…”

“…was produced starting from racemic N -acetyl-Met. Hydroxylation of AAs is one of the numerous theoretical MEC expansion possibilities for the production of oxo-functionalized AAs ( Hibi et al, 2012 ; Smirnov et al, 2012 ; Busto et al, 2014 ; Peters and Buller, 2019 ).…”

“…Other multienzymatic or chemoenzymatic strategies have been proposed for the synthesis of NcAAs, although they have not found yet such a biotechnological interest as those reported above. On the other hand, it is important to bear in mind that besides the more than 20 different enzymes comprised in the different MECs presented in this paper, many enantioselective or stereospecific “free” enzymes are useful for NcAAs synthesis [e.g., proteases ( Nakao et al, 1996 ; Miyazawa, 1999 ), 2-oxoglutarate-dependent oxygenases ( Peters and Buller, 2019 ), aldolases ( Xue et al, 2018 ; Fesko, 2019 ) or other different PLP-enzymes ( Di Salvo et al, 2020 )].…”