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AssigneeGenencor International, Inc.
InventorVirgil Bryan Lawlis
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The invention includes novel fusion DNA sequences encoding fusion polypeptides which when expressed in a filamentous fungus result in increased levels of secretion of the desired polypeptide as compared to the expression and secretion of such polypeptides from filamentous fungi transformed with previously used DNA sequences. The fusion DNA sequences comprise from the 5' terminus four DNA sequences which encode a fusion polypeptide comprising, from the amino to carbonyl-terminus, first, second, third and fourth amino acid sequences. The first DNA sequence encodes a signal peptide functional as a secretory sequence in a first filamentous fungus. The second DNA sequence encodes a secreted polypeptide or portion thereof which is normally secreted from the same filamentous fungus or a second filamentous fungus. The third DNA sequence encodes a cleavable linker polypeptide while the fourth DNA sequence encodes a desired polypeptide. When the fusion DNA sequence is expressed either in the first or second filamentous fungus, increased secretion of the desired polypeptide is obtained as compared to that which is obtained when the desired polypeptide is expressed from DNA sequences encoding a fusion polypeptide which does not contain the second polypeptide normally secreted from either of the filamentous fungi.
A fusion DNA sequence encoding a fusion polypeptide comprising, from the 5' end of said fusion DNA sequence, first, second, third and fourth DNA sequences encoding, from the amino- to carboxy-terminus of said fusion polypeptide, corresponding first, second, third and fourth amino acid sequences, said first DNA sequence encoding a signal peptide functional as a secretory sequence in a first filamentous fungus, said second DNA sequence encoding a mature form of a secreted polypeptide normally secreted from said first or a second filamentous fungus or portion thereof comprising greater than 50% of the amino terminal sequence of said secreted polypeptide, said third DNA sequence encoding a cleavable linker polypeptide and said fourth DNA sequence encoding a desired polypeptide, wherein said first and said second filamentous fungi are selected from the group consisting of Aspergillus, Trichoderma and Neurospora and the expression of said fusion DNA sequence in said first or said second filamentous fungus results in increased secretion of said desired polypeptide as compared to the secretion of said desired polypeptide from said first or said second filamentous fungus when expressed as a second fusion polypeptide encoded by a second fusion DNA sequence comprising only said first, third and fourth DNA sequences.
A fusion polypeptide comprising, from the amino-to carboxy-terminus, first, second, third and fourth amino acid sequences, said first amino acid sequence comprising a signal peptide functional as a secretory sequence in a first filamentous fungus, said second amino acid sequence comprising a mature form of a secreted polypeptide normally secreted from said first or a second filamentous fungus or portion thereof comprising greater than 50% of the amino terminal sequence of said secreted polypeptide, said third amino acid sequence comprising a cleavable linker polypeptide and said fourth amino acid sequence comprising a desired polypeptide, wherein said first and said second filamentous fungi are selected from the group consisting of Aspergillus, Trichoderma and Neurospora and the expression of the fusion DNA sequence encoding said fusion polypeptide in said first or said second filamentous fungus results in increased secretion of said desired polypeptide as compared to the secretion of said desired polypeptide from said first or said second filamentous fungus when expressed from a second fusion DNA sequence encoding a second fusion polypeptide comprising said first, third and fourth amino acid sequences.
A fusion DNA sequence encoding a fusion polypeptide comprising, from the 5' end of said fusion DNA sequence, first, second, third and fourth DNA sequences encoding, from the amino- to carboxy-terminus of said fusion polypeptide, corresponding first, second, third and fourth amino acid sequences, wherein
A fusion DNA sequence according to claim 44 herein said third DNA sequence is selected from the group consisting of the prosequence of bovine chymosin, the prosequence of subtilisin, the prosequence of human immunodeficiency virus protease, and sequences recognized and cleaved by trypsin, Factor Xa, collagenase, clostripin, subtilisin, chymosin, and yeast KEX2 protease.
A fusion polypeptide comprising, from the amino-to carboxy-terminus, first, second, third and fourth amino acid sequences, wherein:
Dependencies are inferred from the claim text.