US 11,981,935 B2
7β-hydroxysteroid dehydrogenase mutants and process for the preparation of ursodeoxycholic acid
Dirk Weuster-Botz, Munich (DE); Michael Braun, Bad Mergentheim-Edelfingen (DE); Arno Aigner, Tuntenhausen (DE); Boqiao Sun, Munich (DE); Christina Kantzow, Munich (DE); Sven Bresch, Munich (DE); Daniel Bakonyi, Cologne (DE); and Werner Hummel, Titz (DE)
Assigned to Pharmazell GmbH, Raubling (DE)
Filed by PharmaZell GmbH, Raubling (DE)
Filed on Jan. 15, 2021, as Appl. No. 17/150,243.
Application 17/150,243 is a division of application No. 15/093,570, filed on Apr. 7, 2016, granted, now 10,954,494.
Application 15/093,570 is a continuation of application No. 13/993,235, abandoned, previously published as PCT/EP2011/073141, filed on Dec. 16, 2011.
Claims priority of application No. 10015726 (EP), filed on Dec. 16, 2010.
Prior Publication US 2021/0139862 A1, May 13, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 33/00 (2006.01); C12N 9/00 (2006.01); C12N 9/02 (2006.01); C12N 9/04 (2006.01); C12P 33/02 (2006.01)
CPC C12N 9/0006 (2013.01) [C12N 9/0008 (2013.01); C12P 33/00 (2013.01); C12P 33/02 (2013.01); C12Y 101/01201 (2013.01)] 8 Claims
 
1. A nucleic acid molecule comprising a nucleotide sequence encoding a 7β-HSDH mutant, which catalyzes at least the stereospecific enzymatic reduction of a 7-ketosteroid to the corresponding 7-hydroxysteroid, wherein the mutant has at least 90% sequence identity to SEQ ID NO:2 and at least one mutation in the sequence motif VMVGRRE corresponding to positions 36 to 42 of SEQ ID NO:2, wherein the mutation is selected from the group consisting of:
a) the single mutation G39X1;
b) the single mutation R40X2;
c) the double mutation G39X1 R40X2
d) the double mutation R40X2 R41X3;
e) the double mutation G39X1 R41X3; and
f) the triple mutation G39X1 R40X2 R41X3; wherein each of X1, X2 and X3 can be the same or different and stands for mutated amino acid residues.