Global Patent Index - EP 1012301 A1

EP 1012301 A1 2000-06-28 - TOTAL SYNTHESIS AND FUNCTIONAL OVEREXPRESSION OF A $i(CANDIDA RUGOSA) LIP1 GENE CODING FOR A MAJOR INDUSTRIAL LIPASE

Title (en)

TOTAL SYNTHESIS AND FUNCTIONAL OVEREXPRESSION OF A $i(CANDIDA RUGOSA) LIP1 GENE CODING FOR A MAJOR INDUSTRIAL LIPASE

Title (de)

TOTALSYNTHESE UND FUNKTIONELLE UBEREXPRESSION EINES CANDIDA RUGOSA LIP1 GENS, DAS FÜR DIE, IN DER INDUSTRIE AM MEISTEN VERWENDETE LIPASE KODIERT

Title (fr)

SYNTHESE TOTALE ET SUREXPRESSION FONCTIONNELLE D'UN GENE CANDIDA RUGOSA LIP1 CODANT POUR UNE LIPASE INDUSTRIELLE MAJEURE

Publication

EP 1012301 A1 (EN)

Application

EP 97940483 A

Priority

NL 9700524 W

Abstract (en)

[origin: WO9914338A1] The dimorphic yeast Candida rugosa has an unusual codon-usage which hampers the functional expression of genes derived from this yeast in a conventional heterologous host. Lipases produced by this yeast are extensively used in industrial bioconversions, but commercial lipase samples contain several different isoforms encoded by the LIP genes family. In a first laborious attempt the LIP1 gene, encoding the major isoform of the C. rugosa lipases (CRLs), was systematically modified by site-directed mutagenesis to gain functional expression in S. cerevisiae. As alternative approach, the gene (1688 bp) was completely synthesised with an optimised nucleotide sequence in terms of heterologous expression in yeast and simplified genetic manipulation. The synthetic gene was functionally overexpressed in Pichia pastoris. The recombinant CRL was produced at high level and purity, accounting for 90-95 % of the secreted proteins. The physical-chemical and catalytic properties of the recombinant lipase were compared with those of a commercial, non-recombinant C. rugosa lipase preparation.

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

C12N 15/55; C12N 1/19; C12N 9/20

IPC 8 full level (invention and additional information)

C12N 9/20 (2006.01); C12N 15/55 (2006.01); C12N 15/67 (2006.01)

CPC (invention and additional information)

C12N 9/20 (2013.01); C12N 15/67 (2013.01)

Citation (search report)

See references of WO 9914338A1

Designated contracting state (EPC)

BE DE DK FR GB NL SE

EPO simple patent family

WO 9914338 A1 19990325; AU 4224997 A 19990405; EP 1012301 A1 20000628

INPADOC legal status


2004-03-03 [18D] DEEMED TO BE WITHDRAWN

- Effective date: 20030818

2003-07-09 [RAP1] TRANSFER OF RIGHTS OF AN EP PUBLISHED APPLICATION

- Owner name: LODERS CROKLAAN B.V.

2003-05-21 [17Q] FIRST EXAMINATION REPORT

- Effective date: 20030407

2000-06-28 [17P] REQUEST FOR EXAMINATION FILED

- Effective date: 20000309

2000-06-28 [AK] DESIGNATED CONTRACTING STATES:

- Kind Code of Ref Document: A1

- Designated State(s): BE DE DK FR GB NL SE