EP 2268803 A2 20110105 - NOVEL 12 ALPHA-HYDROXYSTEROID DEHYDROGENASES, PRODUCTION AND USE THEREOF
Title (en)
NOVEL 12 ALPHA-HYDROXYSTEROID DEHYDROGENASES, PRODUCTION AND USE THEREOF
Title (de)
NEUE 12ALPHA-HYDROXYSTEROIDDEHYDROGENASEN, DEREN HERSTELLUNG UND VERWENDUNG
Title (fr)
NOUVELLES 12ALPHA-HYDROXYSTÉROÏDE-DÉSHYDROGÉNASES, ET PRODUCTION ET LEURS APPLICATIONS
Publication
Application
Priority
- EP 2009002190 W 20090325
- EP 08153330 A 20080326
- EP 09724285 A 20090325
Abstract (en)
[origin: EP2105500A1] 12alpha -Hydroxysteroid dehydrogenase (12alpha -HSDH), obtained from Clostridium sp.and having a molecular weight of 27-30 kD, is new. Independent claims are included for: (1) 12alpha -HSDH-mutant with a modified cosubstrate-utilization; (2) a nucleic acid sequence comprising (a) a coding nucleic acid sequence that encodes a protein having (a1) a fully defined 270 amino acid (SEQ ID NO. 2) sequence given in the specification or a fully defined 257 amino acid (SEQ ID NO. 4) sequence given in the specification, each beginning at position +1 or +2 and a protein having (a2) an amino acid sequence derived from a sequence according to (a1) with a percentage identity of at least 60%, (b) a coding nucleic acid sequence according to SEQ ID NO. 1 (comprising a fully defined 813 base pair sequence given in the specification) or SEQ ID NO. 3 (comprising a fully defined 774 base pair sequence given in the specification), or (c) a coding nucleic acid sequence derived from the nucleic acid sequence according to SEQ ID NO. 1 or 3 with a percentage identity of at least 60%; (3) an expression cassette comprising the nucleic acid sequence under the genetic control of at least one regulative nucleic acid sequence; (4) a vector comprising at least one expression cassette; (5) recombinant microorganisms that carry at least one nucleic acid sequence or at least one expression cassette, transformed with at least one vector; (6) the preparation of 12alpha -HSDH; (7) a method for enzymatic oxidation of 12alpha -hydroxysteroid, comprising converting the hydroxysteroid in the presence of the above 12alpha -HSDH and optionally isolating at least one of the formed oxidation product from the reaction mixture; (8) a method for enzymatic reduction of 12-ketosteroid, comprising converting the ketosteroid in the presence of the above 12alpha -HSDH and optionally isolating at least one of the formed reduction product from the reaction mixture; (9) a bioreactor comprising 12alpha -HSDH in immobilized form; (10) a method for qualitative or quantitative measurement of 12-ketosteroid and/or 12alpha -hydroxysteroid, comprising subjecting the steroids to a redox reaction that is catalyzed by the 12alpha -HSDH in the presence of redox equivalent, measuring the change in the concentration of the redox equivalent and qualitatively or quantitatively measuring the 12-ketosteroid and/or 12alpha -hydroxy steroid; and (11) a method for preparing ursodeoxy cholic acid compounds of formula (I), comprising (aa1) oxidizing a cholic acid compound of formula (II), in the presence of 12alpha -HSDH, to its corresponding 12-keto chenodeoxycholic acid of formula (III), (aa2) subsequently converting (III), by deoxygenation, to chenodeoxycholic acid of formula (IV), (aa3) chemically oxidizing (IV), in position 7, to 7-keto-lithocholic acid of formula (V), (aa4) reducing (V) and (aa5) optionally further purifying the reaction product. R : alkyl, NR 1>R 2>, H, alkali metal ion or N(R 3>) 4 +>; R 1>, R 2> : not defined; R 3> : H or alkyl; and R 1> 1> : H or acyl. [Image] [Image] [Image].
IPC 8 full level
C12N 9/04 (2006.01)
CPC (source: EP US)
C12N 9/0006 (2013.01 - EP US); C12P 33/02 (2013.01 - EP US); C12Y 101/01176 (2013.01 - EP US)
Citation (search report)
See references of WO 2009118176A2
Designated contracting state (EPC)
AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO SE SI SK TR
Designated extension state (EPC)
AL BA RS
DOCDB simple family (publication)
EP 2105500 A1 20090930; AU 2009230728 A1 20091001; AU 2009230728 B2 20140724; CN 102007210 A 20110406; CN 102007210 B 20140730; EP 2268803 A2 20110105; JP 2011526481 A 20111013; KR 20110014980 A 20110214; NZ 587858 A 20120831; US 2011091921 A1 20110421; US 2014147887 A1 20140529; WO 2009118176 A2 20091001; WO 2009118176 A3 20100107; WO 2009118176 A8 20101028
DOCDB simple family (application)
EP 08153330 A 20080326; AU 2009230728 A 20090325; CN 200980110831 A 20090325; EP 09724285 A 20090325; EP 2009002190 W 20090325; JP 2011501147 A 20090325; KR 20107023790 A 20090325; NZ 58785809 A 20090325; US 201313969069 A 20130816; US 93425909 A 20090325