Tassano, Erika, Moore, Charles, Dussauge, Solene, Vargas, Alexandra and Snajdrova, Radka (2022) Discovery of novel Fe(II)/α-ketoglutarate-dependent dioxygenases for oxidation of L-proline. Organic Process Research & Development. ISSN 1083-61601520-586X

Abstract

Genome-mining for novel Fe(II)/α-ketoglutarate-dependent dioxygenases (αKGDs) to expand the enzymatic repertoire in the oxidation of L-proline is reported. Through clustering of genes, we predicted regio- and stereoselectivity in hydroxylation reaction and validated this hypothesis experimentally. Two novel by-products in reactions with BcePH and Ssp5PH were observed, isolated and structure was determined as an epoxide and a 3,4-diol, respectively. Mechanism for formation of epoxide is suggested and validated by using 18O-labelling experiment, that proceeds via cis-3-hydroxylation step first, followed by ring closure. A Biocatalytic step was performed on sub-gram quantities of starting material without any significant condition optimization. The substrate concentration, however, is already up to 40-fold higher than the usually reported titers for P450-mediated hydroxylations, showing the synthetic potential of αKGDs on preparative scale

Item Type:	Article
Keywords:	Biocatalysis, chiral building blocks, C-H functionalization, hydroxylation of amino acids, -ketoglutarate dependent dioxygenases
Date Deposited:	10 Mar 2022 00:45
Last Modified:	10 Mar 2022 00:45
URI:	https://oak.novartis.com/id/eprint/46050