Transformation system in the field of filamentous fungal hosts

• US 7,399,627 B2
• Filed: 03/21/2003
• Issued: 07/15/2008
• Est. Priority Date: 10/06/1998
• Status: Expired due to Fees

• Current Assignee: Danisco USA Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Dyadic International (Usa), Inc.
• Inventors: Bousson, Jean Christophe, Pynnonen, Christine Marie, Van Zeijl, Cornelia Marie Johanna, Parriche, Martine, Burlingame, Richard Paul, Olson, Philip Terry, Sinitsyn, Arkady Panteleimonovich, Punt, Peter Jan, Emalfarb, Mark Aaron
• Primary Examiner(s): Ketter; James S
• Application Number:
  US10/394,568
• Publication Number:
  US 20040002136A1
• Time in Patent Office: 1,943 Days
• Field of Search: None
• US Class Current: 435/254.11
• CPC Class Codes: C12N 15/80 for fungi
• CPC Class Codes: C12N 9/2437 Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
• CPC Class Codes: C12N 9/2462 Lysozyme (3.2.1.17)
• CPC Class Codes: C12N 9/2482 Endo-1,4-beta-xylanase (3.2.1.8)
• CPC Class Codes: C12N 9/58 derived from fungi
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: C12Y 302/01008 Endo-1,4-beta-xylanase (3.2.1.8)
• CPC Class Codes: C12Y 302/01091 Cellulose 1,4-beta-cellobiosidase (3.2.1.91)

Abstract

A novel transformation system in the field of filamentous fungal hosts for expressing and secreting heterologous proteins or polypeptides is described. The invention also covers a process for producing large amounts of polypeptide or protein in an economical manner. The system comprises a transformed or transfected fungal strain of the genus Chrysosporium, more particularly of Chrysosporium lucknowense and mutants or derivatives thereof. It also covers transformants containing Chrysosporium coding sequences, as well expression-regulating sequences of Chrysosporium genes. Also provided are novel fungal enzymes and their encoding sequences and expression-regulating sequences.

Claims

1. A mutant Chrysosporium strain comprising a nucleic acid sequence encoding a polypeptide of interest, said nucleic acid sequence being operably linked to a promoter sequence and optionally a secretion signal sequence, said mutant strain expressing said polypeptide of interest at a higher level than the corresponding non-mutant strain under the same conditions, wherein said mutant derived from Chrysosporium lucknowense is UV13-6 (VKM F-3632 D), NG7C-19 (VKM F-3633 D), or UV18-25 (VKM F-3631 D) or is derived from a Chrysosporium lucknowense strain of C. lucknowense strain C1 (VKM F-3500 D), UV13-6 (VKM F-3632 D), NG7C-19 (VKM F-3633 D), or UV18-25 (VKM F-3631 D).

2. A mutant Chrysosporium strain wherein said mutant is or is derived from a Chrysosporium lucknowense mutant strain selected from the group consisting of UV13-6 (VKM F-3632 D), NG7C-19 (VKM F-3633 D), and UV18-25 (VKM F-3631 D).

3. The mutant Chrysosporium strain of claim 2 wherein said mutant is or is derived from a Chrysosporium lucknowense mutant strain UV13-6 (VKM F-3632 D).

4. The mutant Chrysosporium strain of claim 2 wherein said mutant is or is derived from a Chrysosporium lucknowense mutant strain NG7C-19 (VKM F-3633 D).

5. The mutant Chrysosporium strain of claim 2 wherein said mutant is or is derived from a Chrysosporium lucknowense mutant strain UV18-25 (VKM F-3631 D).

6. The mutant Chrysosporium strain of claim 2, 3, 4 or 5 wherein said mutant strain expresses a polypeptide of interest at a higher level than the corresponding original mutant strain under the same conditions.