Method for improved protein production in filamentous fungi

Abstract

The present invention relates to a method for genetically modifying a filamentous fungus host for improved protein production. The method comprises that a filamentous fungus host is genetically modified to overexpress or to be deficient of specific genes. The invention relates also to the modified hosts. Furthermore, the invention relates to a method for improved production or for producing an improved composition of proteins, such as cellulases, hemicellulases, other proteins involved in the degradation of lignocellulosic material, or other proteins, in a filamentous fungus host.

1 . A method to genetically modify a filamentous fungus host for improved protein production, said method comprising genetically modifying a filamentous fungus host to overexpress with increased amount or activity, or to be deficient with reduced or lacking amount or activity of one or more genes selected from the group consisting of genes tre66966 (SEQ ID NO:1), tre112524 SEQ ID NO:2), tre123668 (SEQ ID NO:3) and tre120120 (SEQ ID NO:5); or of the closest homologue of at least one of said genes in  Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium  or  Penicillium;  or of a fragment or derivative of any of said genes or other nucleotide sequence hybridizing under stringent conditions to at least one of said genes or said homologues, said host being capable of increased or decreased production of cellulase, hemicellulase, other proteins involved in degradation of lignocellulosic material and/or other proteins as compared to the parental strain. 
     
     
         2 . The method according to  claim 1 , wherein the filamentous fungus host is genetically modified to overexpress one or more of the genes selected from the group consisting of genes tre66966 (SEQ ID NO:1), tre112524 SEQ ID NO:2), tre123668 (SEQ ID NO:3) and tre120120 (SEQ ID NO:5); or the closest homologue of at least one of said genes in  Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium  or  Penicillium;  or of a fragment or derivative of any of said genes or other nucleotide sequence hybridizing under stringent conditions to at least one of said genes or said homologues, said host being capable of increased production of cellulase, hemicellulase, other proteins involved in degradation of lignocellulosic material and/or other proteins as compared to the parental strain. 
     
     
         3 . The method according to  claim 1 , wherein the filamentous fungus host is genetically modified to be deficient of one or more of the genes selected from the group comprising genes tre66966 (SEQ ID NO:1), tre112524 SEQ ID NO:2), tre123668 (SEQ ID NO:3) and tre120120 (SEQ ID NO:5); or of the closest homologue of at least one of said genes in  Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium  or  Penicillium;  or of a fragment or derivative of any of said genes or other nucleotide sequence hybridizing under stringent conditions to at least one of said genes or said homologues, said host being capable of decreased production of cellulase, hemicellulase, other proteins involved in degradation of lignocellulosic material and/or other proteins as compared to the parental strain. 
     
     
         4 . The method according to  claim 1 , wherein the host is selected from the group consisting of  Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium  and  Penicillium.    
     
     
         5 . A filamentous fungus host genetically modified to overexpress or to be deficient of one or more genes selected from the group consisting of genes tre66966 (SEQ ID NO:1), tre112524 SEQ ID NO:2), tre123668 (SEQ ID NO:3) and tre120120 (SEQ ID NO:5); or of the closest homologue of at least one of said genes in  Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium  or  Penicillium;  or of a fragment or derivative of any of said genes or other nucleotide sequence hybridizing under stringent conditions to at least one of said genes or said homologues, said host being capable of increased or decreased production of cellulase, hemicellulase, other proteins involved in degradation of lignocellulosic material and/or other proteins as compared to the parental strain. 
     
     
         6 . The host according to  claim 5 , wherein the filamentous fungus host is genetically modified to overexpress one or more genes selected from the group consisting of genes tre66966 (SEQ ID NO:1), tre112524 SEQ ID NO:2), tre123668 (SEQ ID NO:3) and tre120120 (SEQ ID NO:5); or of the closest homologue of at least one of said genes in  Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium  or  Penicillium;  or of a fragment or derivative of any of said genes or other nucleotide sequence hybridizing under stringent conditions to at least one of said genes or said homologues, said host being capable of increased production of cellulase, hemicellulase, other proteins involved in degradation of lignocellulosic material and/or other proteins as compared to the parental strain. 
     
     
         7 . The host according to  claim 5 , wherein the filamentous fungus host is genetically modified to be deficient of one or more genes selected from the group consisting of genes tre66966 (SEQ ID NO:1), tre112524 SEQ ID NO:2), tre123668 (SEQ ID NO:3) and tre120120 (SEQ ID NO:5); or of the closest homologue of at least one of said genes in  Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium  or  Penicillium;  or of a fragment or derivative of any of said genes or other nucleotide sequence hybridizing under stringent conditions to at least one of said genes or said homologues, said host being capable of decreased production of cellulase, hemicellulase, other proteins involved in degradation of lignocellulosic material and/or other proteins as compared to the parental strain. 
     
     
         8 . The host according to  claim 5 , wherein the host is selected from the group consisting of  Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium  and  Penicillium.    
     
     
         9 . The host according to  claim 5 , wherein the host is a filamentous fungus production host. 
     
     
         10 . A method for improved production or for producing an improved composition of proteins in a filamentous fungus host, said method comprising a step of cultivating the modified filamentous fungus host of  claim 5  under suitable culture conditions for protein production. 
     
     
         11 . The method according to  claim 10 , wherein the protein is selected from the group consisting of cellulases, hemicellulases, side chain cleaving enzymes, lignocellulose degrading enzymes, pectinases, ligninases; amylolytic enzymes; proteases; invertases; phytases, phosphatases, and hydrophobins. 
     
     
         12 . The method according to  claim 10 , wherein the protein is a heterologous or recombinant protein and expressed under a promoter of a gene encoding any one of the proteins whose production is effected by genetical modification of the host. 
     
     
         13 . The method according to  claim 12 , wherein the the promoter is a promoter of a gene encoding a cellulase, a hemicellulase, other protein involved in the degradation of lignocellulosic material or other secreted protein. 
     
     
         14 . The method according to  claim 12 , wherein t the promoter is a promoter of a gene encoding a protein selected from the group consisting of cellulases, hemicellulases, side chain cleaving enzymes, lignocellulose degrading enzymes, pectinases, ligninases; amylolytic enzymes; proteases; invertases; phytases, phosphatases and hydrophobins.