Title:
METHOD OF GENERATING DIVERSITY INTO LIPOLYTIC ENZYMES AND LIPOLYTIC ENZYME GENES
Document Type and Number:
WIPO Patent Application WO2002066622
Kind Code:
A3
Abstract:
The inventors have isolated novel genes with a high homology to the T. lanuginosus lipase gene and are thus well suited for use in gene shuffling. Accordingly, the invention provides a method of generating genetic diversity into lipolytic enzymes by family shuffling of two or more homologous genes which encode lipolytic enzymes. The DNA shuffling technique is used to create a library of shuffled genes, and this is expressed in a suitable expression system and the expressed proteins are screened for lipolytic enzyme activity. The expressed proteins may further be screened to identify lipolytic enzymes with improved properties. The invention also provides a polynucleotide comprising a nucleotide sequence encoding a lipolytic enzyme and a lipolytic enzyme (a polypeptide with lipolytic enzyme activity).
Inventors:
TSUTSUMI NORIKO (JP)
VIND JESPER (DK)
PATKAR SHAMKANT ANANT (DK)
VIND JESPER (DK)
PATKAR SHAMKANT ANANT (DK)
Application Number:
PCT/DK2002/000124
Publication Date:
January 08, 2004
Filing Date:
February 25, 2002
Export Citation:
Assignee:
NOVOZYMES AS (DK)
TSUTSUMI NORIKO (JP)
VIND JESPER (DK)
PATKAR SHAMKANT ANANT (DK)
TSUTSUMI NORIKO (JP)
VIND JESPER (DK)
PATKAR SHAMKANT ANANT (DK)
International Classes:
C12N15/09; A21D8/04; A23L7/104; C11D3/386; C12N1/15; C12N1/19; C12N1/21; C12N5/10; C12N9/20; C12N15/10; (IPC1-7): C12N9/20; C12N15/10
Domestic Patent References:
| WO1997007205A1 | 1997-02-27 | |||
| WO1997007206A1 | 1997-02-27 | |||
| WO1998041622A1 | 1998-09-24 | |||
| WO2000032758A1 | 2000-06-08 |
Foreign References:
| US6159687A | 2000-12-12 |
Other References:
DATABASE EMBL [online] XP002958619, retrieved from EMBL Database accession no. A32008
DATABASE EMBL [online] 6 April 1998 (1998-04-06), MULLER S ET AL, XP002958618, retrieved from EMBL Database accession no. AF054513
DATABASE EMBL [online] XP002958620, retrieved from EMBL Database accession no. AR083396
DATABASE EBI [online] XP002958621, retrieved from GPOP Database accession no. AAE05236
DATABASE EBI [online] XP002958622, retrieved from GSP Database accession no. AAR81990
KIM MYUNG HEE ET AL: "Substitution of glycine 275 by glutamate (G275E) in lipase of Bacillus Stearothermophilus affects its catalytic activity and enantio-and chain length specificity.", JOURNAL OF MICROBIOLOGY AND BIOTECNOLOGY, vol. 10, no. 5, 2000, pages 764 - 769, XP002902706
CRAMERI A ET AL: "DNA shuffling of a family of genes from diverse species accelerates directed evolution", NATURE, MACMILLAN JOURNALS LTD. LONDON, GB, vol. 391, 15 January 1998 (1998-01-15), pages 288 - 291, XP002178865, ISSN: 0028-0836
DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1978, OSO B A: "THE LIPASE ACTIVITY OF TALAROMYCES-EMERSONII", XP002958623, Database accession no. PREV197967018106
DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; December 1996 (1996-12-01), ADEKUNLE A A ET AL: "Lipase activity of fourteen fungi on cucumeropsis mannii seeds.", XP002958624, Database accession no. PREV199900188922
DATABASE EMBL [online] 6 April 1998 (1998-04-06), MULLER S ET AL, XP002958618, retrieved from EMBL Database accession no. AF054513
DATABASE EMBL [online] XP002958620, retrieved from EMBL Database accession no. AR083396
DATABASE EBI [online] XP002958621, retrieved from GPOP Database accession no. AAE05236
DATABASE EBI [online] XP002958622, retrieved from GSP Database accession no. AAR81990
KIM MYUNG HEE ET AL: "Substitution of glycine 275 by glutamate (G275E) in lipase of Bacillus Stearothermophilus affects its catalytic activity and enantio-and chain length specificity.", JOURNAL OF MICROBIOLOGY AND BIOTECNOLOGY, vol. 10, no. 5, 2000, pages 764 - 769, XP002902706
CRAMERI A ET AL: "DNA shuffling of a family of genes from diverse species accelerates directed evolution", NATURE, MACMILLAN JOURNALS LTD. LONDON, GB, vol. 391, 15 January 1998 (1998-01-15), pages 288 - 291, XP002178865, ISSN: 0028-0836
DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1978, OSO B A: "THE LIPASE ACTIVITY OF TALAROMYCES-EMERSONII", XP002958623, Database accession no. PREV197967018106
DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; December 1996 (1996-12-01), ADEKUNLE A A ET AL: "Lipase activity of fourteen fungi on cucumeropsis mannii seeds.", XP002958624, Database accession no. PREV199900188922
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