Fungal production system
The present invention provides a new fungal production system comprising a fungal host strain of Chrysosporium lucknowense wherein the endogenous cellulase secretion is less than 20% of the endogenous cellulase secretion of Chrysosporium lucknowense strain UV 18-25. Preferably, also the secretion of endogenous protease, endogenous β-glucanase and endogenous cellobiohydrolase is less than 20% of the secretion of Chrysosporium lucknowense strain UV 18-25. Furthermore, fungal host strains are provided wherein several genes have been disrupted. According to another aspect of the invention a method for homologous and/or heterologous production of a pure protein with a purity of higher than 75%, comprising expressing a gene encoding said protein in the strains according to the invention have been described. Furthermore, a method for production of artificial protein mixes comprising expressing a gene encoding each of said proteins in a strain according to the invention have been disclosed. Finally a method for simplified screening of strains functionally expressing a desired enzyme by application of said strains have been provided.
1. A chimeric gene comprising:
(i) the nucleotide sequence of SEQ ID NO:26, or
(ii) a transcriptionally active part of the nucleotide sequence of SEQ ID NO:26 that comprises the nucleotide sequence of SEQ ID NO:25.
2. A fungal host cell comprising the chimeric gene of claim 1 .
3. The host cell of claim 2 , wherein said host cell is a Chrysosporium lucknowense C1 cell or a derivative thereof.
4. The host cell of claim 3 , wherein said host cell is selected from the group consisting of:
(a) Chrysosporium lucknowense C1 strain UV18-25 deposited in the All Russian Collection Microorganisms (VKM) as Deposit No. VKM F3631 D, and
(b) a derivative of said Chrysosporium lucknowense C1 strain of (a).
5. The host cell of claim 4 , wherein said host cell is selected from the group consisting of:
(a) W1L, deposited at CBS under accession number 122189, and
(b) W1L #100.1, deposited at CBS under accession number 122190.
6. A method for homologous or heterologous production of a protein comprising expressing the chimeric gene of claim 1 in a fungal host cell, thereby producing said protein.
7. The method of claim 6 , wherein the said host cell is a Chrysosporium lucknowense C1 cell or a derivative thereof.
8. The method of claim 7 , wherein the said host cell is selected from the group consisting of:
(a) W1L, deposited at CBS under accession number 122189, and
(b) W1L #100.1, deposited at CBS under accession number 122190.
9. The chimeric gene of claim 1 , wherein said chimeric gene comprises the nucleotide sequence of SEQ ID NO:26.