NOVEL EXPRESSION-REGULATING SEQUENCES AND EXPRESSION PRODUCTS IN THE FIELD OF FILAMENTOUS FUNGI

• US 20130143271A1
• Filed: 03/14/2011
• Published: 06/06/2013
• Est. Priority Date: 10/06/1998
• Status: Abandoned Application

• Current Assignee: Dyadic International (Usa), Inc.
• Original Assignee: Dyadic International (Usa), Inc.
• Inventors: Emalfarb, Mark Aaron, Punt, Peter Jan, Van Zeijl, Cornelia Maria Johanna
• Application Number:
  US13/046,772
• Publication Number:
  US 20130143271A1
• Time in Patent Office: Days
• US Class Current: 435/72
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12N 9/24 acting on glycosyl compounds (3.2)
• 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 9/248 Xylanases
• 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

The invention pertains to novel proteins corresponding to Chrysosporium glycosyl hydrolases of families 7 and 10, exhibiting a minimum aminoacid identity of 70 and 75%, respectively, with the amino acid sequence of SEQ ID No'"'"'s 2 and 4, and to a protein corresponding to a Chrysosporium glyceraldehyde phosphate dehydrogenase, exhibiting at least 86% amino acid identity with the partial amino acid sequence of SEQ ID No. 6. The invention further relates to nucleic acid sequences encoding these proteins, and especially to promoter sequences regulating the expression of the corresponding genes. The preferred host for expressing these genes is a fungus, especially a Chrysosporium strain.

Claims

1. A protein corresponding to a Chrysosporium glycosyl hydrolase family 7, exhibiting at least 75% amino acid identity [as determined by BLAST algorithm] with the amino acid sequence of SEQ ID No.2 or a part thereof having at least 20 contiguous amino acids which are identical to the corresponding part of the amino acid sequence 1-246 or 394-526 of SEQ ID No.2.

3. A process of hydrolysing β

-glucosidic bonds, comprising the use of an enzyme according to claim 1.

4. A nucleic acid sequence encoding a protein according to claim 1.

2. A protein corresponding to a Chrysosporium glyceraldehyde phosphate dehydrogenase, exhibiting at least 86% amino acid identity [as determined by BLAST algorithm] with the partial amino acid sequence of SEQ ID No. 6 or a part thereof having at least 20 contiguous amino acids which are identical to the corresponding part of the amino acid sequence 1-277 of SEQ ID No. 6.

5. A nucleic acid sequence encoding a protein according to claim 2.

6. A nucleic acid sequence comprising at least 70% of the nucleotides contained in the 5′

-non-coding region of the nucleic acid sequence of any one of SEQ ID No'"'"'s 1 and 5.

7. A nucleic acid construct comprising a nucleic acid expression-regulatory region derived from Chrysosporium, contained in the 5′

-noncoding region of the nucleic acid sequence of any one of SEQ ID No'"'"'s 1 and 5, operationally linked to a nucleic acid sequence encoding a polypeptide of interest.

8. A recombinant microbial strain, preferably a fungal strain, containing a nucleic acid sequence according to claim 7, and capable of expressing the polypeptide encoded by the coding nucleic acid sequence.

9. A recombinant microbial strain, preferably a fungal strain, containing a nucleic acid sequence according to claim 8, and capable of expressing the polypeptide encoded by the coding nucleic acid sequence.

10. A recombinant microbial strain, preferably a fungal strain, containing a nucleic acid sequence according to claim 9, and capable of expressing the polypeptide encoded by the coding nucleic acid sequence.

11. A recombinant microbial strain, preferably a fungal strain, containing a nucleic acid sequence according to claim 10, and capable of expressing the polypeptide encoded by the coding nucleic acid sequence.

12. A process of producing a polypeptide using a construct according to claim 10.

13. A process of producing a polypeptide using a microbial strain according to claim 11.

14. An oligonucleotide probe comprising at least 15 contiguous nucleotides of the nucleic acid sequence of any one of SEQ ID No'"'"'s 1 and 5, or its complement.

15. An oligonucleotide probe according to claim 14, wherein the probe is 20-50 nucleotides in length.

16. An oligonucleotide probe according to claim 14 wherein the probe is labeled with a detectable label.