Global Patent Index - EP 1002097 A2

EP 1002097 A2 2000-05-24 - DEHYDROGENASES WITH IMPROVED NAD-DEPENDENCE, THE PRODUCTION THEREOF, AND THEIR USE

Title (en)

DEHYDROGENASES WITH IMPROVED NAD-DEPENDENCE, THE PRODUCTION THEREOF, AND THEIR USE

Title (de)

DEHYDROGENASEN MIT VERBESSERTER NAD-ABHÄNGIGKEIT, DEREN HERSTELLUNG UND VERWENDUNG

Title (fr)

DESHYDROGENASES A DEPENDANCE NAD AMELIOREE, LEUR PRODUCTION ET LEUR UTILISATION

Publication

EP 1002097 A2 (DE)

Application

EP 99923384 A

Priority

  • DE 9900848 W
  • DE 19812004 A

Abstract (en)

[origin: DE19812004A1] The NADH specificity of preferred NADPH-dependent dehydrogenases can be improved by basicity reduction in the coenzyme docking area through corresponding genetically engineered modification of the relevant amino acid sequence. Dehydrogenases with NADH-dependence suitable for preparative purposes which correspond to a kcat/KM value for NAD<+>>/=20 can be microbially obtained. To this end, a microorganism is used which codes for the enzyme and which has a gene sequence in the genetic code. The gene sequence codes for the enzyme with an amino acid sequence whose basic amino acid(s) are at least partially replaced on the coenzyme binding site or sites by uncharged amino acids. Alternatively or additionally, basic or positively charged or uncharged amino acid(s) can be replaced by negatively charged amino acid(s). The inventive method is especially used for obtaining short-chain dehydrogenases with coenzyme binding sites on the N-terminus. According to the example, alcohol-dehydrogenases were obtained with the aid of E.coli HB101+ (pUBS 520) which contains the mutated gene in an overexpressed manner. Said gene codes for the dehydrogenase and is cloned in the expression plasmi pKK-117-3HB.

IPC 1-7 (main, further and additional classification)

C12N 15/53; C12N 9/04; C12P 7/02

IPC 8 full level (invention and additional information)

C12N 15/09 (2006.01); C12N 9/04 (2006.01); C12N 9/14 (2006.01); C12N 15/53 (2006.01); C12P 7/02 (2006.01); C12P 41/00 (2006.01); C12R 1/19 (2006.01)

CPC (invention and additional information)

C12N 9/0006 (2013.01); C12P 7/02 (2013.01)

Designated contracting state (EPC)

AT CH DE DK ES FR GB IT LI NL SE

EPO simple patent family

DE 19812004 A1 19990930; EP 1002097 A2 20000524; JP 2001526547 A 20011218; US 6413750 B1 20020702; WO 9947684 A2 19990923; WO 9947684 A3 20000323

INPADOC legal status


2007-07-11 [18D] DEEMED TO BE WITHDRAWN

- Effective date: 20070117

2006-10-11 [17Q] FIRST EXAMINATION REPORT

- Effective date: 20050203

2005-03-23 [17Q] FIRST EXAMINATION REPORT

- Effective date: 20050203

2002-07-10 [RAP1] TRANSFER OF RIGHTS OF AN EP PUBLISHED APPLICATION

- Owner name: FORSCHUNGSZENTRUM JUELICH GMBH

2000-05-24 [17P] REQUEST FOR EXAMINATION FILED

- Effective date: 19991211

2000-05-24 [AK] DESIGNATED CONTRACTING STATES:

- Kind Code of Ref Document: A2

- Designated State(s): AT CH DE DK ES FR GB IT LI NL SE