	US 11,965,199 B2
	21-hydroxylation of steroids
Claus Lattemann, Frankfurt am Main (DE); Thomas Stillger, Frankfurt am Main (DE); Bernd Janocha, Frankfurt am Main (DE); Hans-Falk Rasser, Frankfurt am Main (DE); Sebastian Rissom, Frankfurt am Main (DE); Simone Anderko, Saarbrücken (DE); Rita Bernhardt, Saarbrücken (DE); and Frank Hannemann, Saarbrücken (DE)
Assigned to SANOFI, Paris (FR)
Filed by EUROAPI, Paris (FR)
Filed on Jan. 31, 2022, as Appl. No. 17/589,667.
Application 16/453,651 is a division of application No. 15/523,107, granted, now 10,385,376, issued on Aug. 20, 2019, previously published as PCT/EP2015/075096, filed on Oct. 29, 2015.
Application 17/589,667 is a continuation of application No. 16/453,651, filed on Jun. 26, 2019, granted, now 11,608,514.
Claims priority of application No. 14306740 (EP), filed on Oct. 30, 2014.
Prior Publication US 2022/0228189 A1, Jul. 21, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 33/06 (2006.01)

CPC C12P 33/06 (2013.01)

20 Claims

1. A process for the hydroxylation of the carbon atom 21 of a steroid, comprising the steps of:

(a) providing a cell expressing:

(i) a heterologous CYP21A2 protein or a functional variant thereof;

(ii) at least one heterologous electron transfer system capable of transferring electrons to CYP21A2, wherein the at least one electron transfer system is selected from the group consisting of:

the combination of a flavodoxin reductase (Fpr) and an adrenodoxin;

the combination of a Fpr and the ferredoxin domain of an electron transfer domain (etp^fd);

the combination of an adrenodoxin reductase homolog (Arh) and an adrenodoxin; and

the combination of an Arh and etp^fd;and

(iii) one or more chaperones facilitating folding of CYP21A2; and

(b) adding the steroid to the cell.