21-hydroxylation of steroids
AssigneeEuroapi
InventorsClaus Lattemann, Thomas Stillger, Bernd Janocha, Hans-Falk Rasser, Sebastian Rissom, Simone Anderko, Rita Bernhardt, Frank Hannemann
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Abstract
Generally, the present invention relates to the field of steroid hydroxylation. More specifically, the present invention relates to a method for the 21-hydroxylation of steroids in cells. It also relates to cells expressing a steroid 21-hydroxylating enzyme or steroid 21-hydroxylase, expression vectors comprising a nucleic acid encoding for a steroid 21-hydroxylase and a kit for carrying out the method for the 21-hydroxylation of steroids in cells.
Claims18
A process for the hydroxylation of the carbon atom 21 of a steroid, comprising the steps of:
- (a) providing a prokaryotic 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;
- and
- (iii) one or more chaperones facilitating folding of CYP21A2;
- and
- (b) adding the steroid to the prokaryotic cell.
A prokaryotic 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, and
- (iii) one or more chaperones facilitating folding of CYP21A2.
A kit for the hydroxylation of the carbon atom 21 of a steroid, comprising a prokaryotic cell expressing:
- (a) a heterologous CYP21A2 protein or a functional variant thereof;
- (b) at least one heterologous electron transfer system capable of transferring electrons to CYP21A2;
- and
- (c) one or more chaperones facilitating folding of CYP21A2;
- wherein the prokaryotic cell is an E. coli cell;
- and
- wherein the prokaryotic cell further expresses a heterologous gene encoding for an enzyme catalyzing a step in the heme biosynthesis pathway.
Dependencies are inferred from the claim text.