Abstract: The present invention relates to isolated fungal promoter DNA sequences, to DNA constructs, vectors, and fungal host cells comprising these promoters in operative association with coding sequences encoding polypeptides. The present invention also relates to methods for expressing a gene and/or producing a polypeptide using the new promoters isolated. The present invention also relates to methods for altering the transcription level and/or regulation of an endogenous gene using the new promoter of the invention.

Abstract: The present invention relates to isolated fungal promoter DNA sequences, to DNA constructs, vectors, and fungal host cells comprising these promoters in operative association with coding sequences encoding polypeptides. The present invention also relates to methods for expressing a gene and/or producing a polypeptide using the new promoters isolated. The present invention also relates to methods for altering the transcription level and/or regulation of an endogenous gene using the new promoter of the invention.

Abstract: The present invention relates to a process for the production of a recombinant polypeptide of interest, a polypeptide obtained by said process, a recombinant polynucleotide, an expression vector, an expression construct and to the use of a specific signal peptide and of a polynucleotide encoding said specific signal peptide for the production of a recombinant polypeptide of interest.

Abstract: The present invention relates to a method for improving protein yield. The method comprises modifying the value of a set of relevant protein features to fall within an optimal range or to become more close to an optimal value for one or more protein features in the eukaryotic host.

Abstract: The present invention relates to functional cDNA and genomic sequences encoding PrtT proteins, which have transcriptional activity on a protease promoter, to PrtT proteins and to their use. The invention further relates to two distinct types of filamentous fungal cells. Filamentous fungal cells are transformed to over-express these PrtT proteins: this type of filamentous fungus will be highly suited as protease producer. Alternatively, the endogenous prtT genes of filamentous fungal cells are inactivated: this type of filamentous fungus is highly suited for the production of any polypeptide native or not which is highly sensitive for protease degradation. The PrtT proteins of the invention provide means for identification of functional homologues in other species.

Abstract: The present invention relates to polypeptides that have an activity corresponding to at least one activity of the SEC61 polypeptide, polynucleotides encoding these polypeptides and the use thereof in the preparation of host cells suitable for production of a polypeptide of interest. Such host cells may have an increased capacity to secrete a polypeptide of interest.

Abstract: The invention relates to a nucleotide sequence comprising; a synonymous nucleotide coding sequence with optimized codon frequency such that a native codon has been exchanged with a synonymous codon, the synonymous codon encoding the same amino acid as the native codon and having a higher frequency in codon usage as defined in Table 1 than the native codon; and optionally the nucleotide sequence comprises control sequences such as; one translational termination sequence orientated in 5? towards 3? direction selected from the following list of sequences; TAAG, TAGA and TAAA, preferably TAAA, and/or one translational initiator coding sequence orientated in 5? towards 3? direction selected from the following list of sequences; gctnccyyc (SEQ ID NO:20), using ambiguity codes for nucleotides; v (A/C/G); n (A/C/G/T), preferably 5?-GCT TCC TTC-3? (SEQ ID NO:21).

Abstract: The present invention relates to functional cDNA and genomic sequences encoding PrtT proteins, which have transcriptional activity on a protease promoter, to PrtT proteins and to their use. The invention further relates to two distinct types of filamentous fungal cells. Filamentous fungal cells are transformed to over-express these PrtT proteins: this type of filamentous fungus will be highly suited as protease producer. Alternatively, the endogenous prtT genes of filamentous fungal cells are inactivated: this type of filamentous fungus is highly suited for the production of any polypeptide native or not which is highly sensitive for protease degradation. The PrtT proteins of the invention provide means for identification of functional homologues in other species.

Abstract: We describe a recombinant host cell for the production of a compound of interest as well as isolated fungal promoter DNA sequences, to DNA constructs, vectors, and fungal host cells comprising these promoters in operative association with coding sequences encoding a compound of interest. We also describe methods for expressing a gene of interest and/or producing compounds of interest using a promoter according to the invention.

Abstract: The present invention relates to a method to improve the secretion of a protein of interest by a filamentous fungal cell comprising inducing a phenotype in the cell selected from the group consisting of a lowered ERAD, an elevated UPR that does not induce an elevated ERAD, wherein ERAD preferably is lowered. The invention further relates to the filamentous fungal cell comprising the phenotype described above. The invention also relates to polynucleotides and polypeptides whose expression can be modulated in the filamentous fungal cell to obtain the above-described phenotype.

Abstract: The present invention relates to methods of optimization of a protein coding sequences for expression in a given host cell. The methods apply genetic algorithms to optimise single codon fitness and/or codon pair fitness sequences coding for a predetermined amino acid sequence. In the algorithm generation of new sequence variants and subsequent selection of fitter variants is reiterated until the variant coding sequences reach a minimum value for single codon fitness and/or codon pair fitness. The invention also relates to a computer comprising a processor and memory, the processor being arranged to read from and write into the memory, the memory comprising data and instructions arranged to provide the processor with the capacity to perform the genetic algorithms for optimisation of single codon fitness and/or codon pair fitness.

Abstract: We have discovered DNA binding sites which are specifically recognized by PrtT, a transcriptional activator for protease genes. The DNA binding site can be defined structurally by a consensus nucleotide sequence and functionally by PrtT's ability to regulate transcriptional activation through that sequence. Both PrtT and its cognate DNA binding site (i.e., the nucleotide sequence in each promoter that is recognized by PrtT) can be used in a gene expression system. Possession of only a PrtT transcriptional activator is insufficient, its cognate DNA binding site is necessary for recognition by PrtT (i.e., binding to the site and activating transcription under appropriate conditions). A functional site, such as one obtained from a wild-type fungal gene, will confer PrtT-dependent transcriptional activation on 3?-downstream sequences. A mutation of a wild-type promoter that results in a non-functional site will abolish PrtT-dependent transcriptional activation of 3?-downstream sequences.

Abstract: The present invention relates to isolated fungal promoter DNA sequences, to DNA constructs, vectors, and fungal host cells comprising these promoters in operative association with coding sequences encoding polypeptides. The present invention also relates to methods for expressing a gene and/or producing a polypeptide using the new promoters isolated. The present invention also relates to methods for altering the transcription level and/or regulation of an endogenous gene using the new promoter of the invention.

Abstract: The present invention relates to isolated promoter DNA sequences, to DNA constructs, vectors, and host cells comprising these promoters in operative association with coding sequences. The present invention also relates to methods for expressing a gene and/or producing a biological compound using the new promoters isolated. The present invention also relates to methods for altering the transcription level and/or regulation of an endogenous gene using the new promoter of the invention.

Abstract: The invention relates to a nucleotide sequence comprising: a synonymous nucleotide coding sequence with optimized codon frequency such that a native codon has been exchanged with a synonymous codon, said synonymous codon encoding the same amino acid as the native codon and having a higher frequency in codon usage as defined in Table 1 than the native codon; and optionally said nucleotide sequence comprises control sequences such as: one translational termination sequence orientated in 5? towards 3? direction selected from the following list of sequences: TAAG, TAGA and TAAA, preferably TAAA, and/or one translational initiator coding sequence orientated in 5? towards 3? direction selected from the following list of sequences: gctnccyyc, using ambiguity codes for nucleotides: v (A/C/G); n (A/C/G/T), preferably 5?-GCT TCC TTC-3?.

Abstract: A nucleotide sequence is provided which encodes a peptide having &bgr;-xylosidase activity and exhibits at least 30% amino acid identity with the amino acid sequence shown in SEQ ID NO. 1 and described in SEQ ID NO. 3 or hybridises under stringent conditions with a nucleotide sequence shown in SEQ ID NO. 1, or a part thereof having at least 15 nucleotides encoding an amino acid sequence shown in SEQ ID NO. 1 and described in SEQ ID NO. 3. Also provided is a peptide having &bgr;-xylosidase activity and exhibiting at least 30% amino acid identity with the amino acid sequence shown in SEQ ID NO. 1 and described in SEQ ID NO. 3 or a part thereof having at least 8 amino acids shown in SEQ ID NO. 1 and described in SEQ ID NO. 3.