US2009275079A1PendingUtilityA1

Novel genes encoding novel proteolytic enzymes

52
Assignee: DSM IP ASSETS BVPriority: Dec 21, 2001Filed: Oct 25, 2007Published: Nov 5, 2009
Est. expiryDec 21, 2021(expired)· nominal 20-yr term from priority
Inventors:Luppo EdensAlard Van Albertus DijkPhillip KrubasikKaj AlbermannAlexander StockErik KimpelSabrine KlaubauerChristian WagnerAndreas FritzWilk Von GustedtOliver HenrichDieter MaierFabio SpreaficoUlrike FolkersSylvia HopperWolfram KemmnerPamela TanJosephine StieblerRicahrd Albang
C12N 9/62G01N 2333/38G01N 33/56961C07K 2319/00G01N 33/573C12Q 1/37
52
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

The invention relates to newly identified gene sequences that encode novel proteases obtainable from Aspergillus niger . The invention features the full length gene sequence of the novel genes, their cDNA sequences as well as the full-length functional protein and fragments thereof. The invention also relates to methods of using these enzymes in industrial processes and methods of diagnosing fungal infections. Also included in the invention are cells transformed with DNA according to the invention and cells wherein a protease according to the invention is genetically modified to enhance or reduce its activity and/or level of expression.

Claims

exact text as granted — not AI-modified
1 . An isolated polynucleotide that encodes a tripeptidyl peptidase wherein said polynucleotide hybridizes to the full length complement of a nucleotide sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:8; SEQ ID NO:11; SEQ ID NO:13 to SEQ ID NO:25; SEQ ID NO:27 to SEQ ID NO:34; SEQ ID NO:36 to SEQ ID NO:49; SEQ ID NO:51 to SEQ ID NO:57; SEQ ID NO:58 to SEQ ID NO:65; SEQ ID NO:68; SEQ ID NO:70 to SEQ ID NO:82; SEQ ID NO:84 to SEQ ID NO:91; SEQ ID NO:93 to SEQ ID NO:106; and SEQ ID NO:108 to SEQ ID NO:115 under hybridization conditions of 5×SSC, 5×Denhardt's solution, and 1.0% SDS at 68° C. and wash conditions of 0.2×SSC and 0.1% SDS at room temperature. 
     
     
         2 . The isolated polynucleotide of  claim 1 , which comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:8; SEQ ID NO:11; SEQ ID NO:13 to SEQ ID NO:25; SEQ ID NO:27 to SEQ ID NO:34; SEQ ID NO:36 to SEQ ID NO:49; SEQ ID NO:51 to SEQ ID NO:57; SEQ ID NO:58 to SEQ ID NO:65; SEQ ID NO:68; SEQ ID NO:70 to SEQ ID NO:82; SEQ ID NO:84 to SEQ ID NO:91; SEQ ID NO:93 to SEQ ID NO:106; and SEQ ID NO:108 to SEQ ID NO:115. 
     
     
         3 . The isolated polynucleotide of  claim 1  obtainable from a filamentous fungus. 
     
     
         4 . The isolated polynucleotide of  claim 3  obtainable from  Aspergillus niger.    
     
     
         5 . An isolated polynucleotide encoding a polypeptide having tripeptidyl peptidase-activity, which is at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:115 to SEQ ID NO:122; SEQ ID NO:125; SEQ ID NO:127 to SEQ ID NO:139; SEQ ID NO:141 to SEQ ID NO:148; SEQ ID NO:150 to SEQ ID NO:163; and SEQ ID NO:165 to SEQ ID NO:171. 
     
     
         6 . The isolated polynucleotide of  claim 7  wherein said polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:115 to SEQ ID NO:122; SEQ ID NO:125; SEQ ID NO:127 to SEQ ID NO:139; SEQ ID NO:141 to SEQ ID NO:148; SEQ ID NO:150 to SEQ ID NO:163; and SEQ ID NO:165 to SEQ ID NO:171. 
     
     
         7 . The isolated polynucleotide of  claim 1 , which hybridizes to the full length of a nucleotide sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:8; SEQ ID NO:11; SEQ ID NO:13 to SEQ ID NO:25; SEQ ID NO:27 to SEQ ID NO:34; SEQ ID NO:36 to SEQ ID NO:49; SEQ ID NO:51 to SEQ ID NO:57; SEQ ID NO:58 to SEQ ID NO:65; SEQ ID NO:68; SEQ ID NO:70 to SEQ ID NO:82; SEQ ID NO:84 to SEQ ID NO:91; SEQ ID NO:93 to SEQ ID NO:106; and SEQ ID NO:108 to SEQ ID NO:115. 
     
     
         8 . A vector comprising the polynucleotide sequence of  claim 1 . 
     
     
         9 . The vector of  claim 8  wherein said polynucleotide sequence is operatively linked with regulatory sequences suitable for expression of said polynucleotide sequence in a suitable host cell. 
     
     
         10 . The vector of  claim 9  wherein said suitable host cell is a filamentous fungus. 
     
     
         11 . An isolated recombinant host cell comprising the vector of  claim 9 . 
     
     
         12 . A method to prepare a tripeptidyl peptidase comprising the steps of culturing a host cell comprising the vector of  claim 9  and isolating said tripeptidyl peptidase from said host cell. 
     
     
         13 . A vector comprising the polynucleotide sequence of  claim 5 . 
     
     
         14 . The vector of  claim 13  wherein said polynucleotide sequence is operatively linked with regulatory sequences suitable for expression of said polynucleotide sequence in a suitable host cell. 
     
     
         15 . The vector of  claim 14  wherein said suitable host cell is a filamentous fungus. 
     
     
         16 . An isolated recombinant host cell comprising the vector of  claim 14 . 
     
     
         17 . A method to prepare a tripeptidyl peptidase comprising the steps of culturing a host cell comprising the vector of  claim 14  and isolating said tripeptidyl peptidase from said host cell. 
     
     
         18 . An isolated polypeptide having tripeptidyl peptidase activity, which comprises an amino acid sequence at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:115 to SEQ ID NO:122; SEQ ID NO:125; SEQ ID NO:127 to SEQ ID NO:139; SEQ ID NO:141 to SEQ ID NO:148; SEQ ID NO:150 to SEQ ID NO:163; and SEQ ID NO:165 to SEQ ID NO:171. 
     
     
         19 . The isolated polypeptide of  claim 18  wherein said polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:115 to SEQ ID NO:122; SEQ ID NO:125; SEQ ID NO:127 to SEQ ID NO:139; SEQ ID NO:141 to SEQ ID NO:148; SEQ ID NO:150 to SEQ ID NO:163; and SEQ ID NO:165 to SEQ ID NO:171. 
     
     
         20 . An isolated polypeptide obtainable by expressing the polynucleotide of  claim 1  in a host cell. 
     
     
         21 . An isolated polypeptide obtainable by expressing the polynucleotide of  claim 5  in a host cell. 
     
     
         22 . An isolated recombinant host cell expressing the polypeptide of  claim 18 . 
     
     
         23 . A fusion protein comprising the amino acid sequence according to  claim 18 . 
     
     
         24 . A fusion protein comprising the amino acid sequence of the polypeptide according to  claim 20 . 
     
     
         25 . A fusion protein comprising the amino acid sequence of the polypeptide according to  claim 21 .

Cited by (0)

No later patents cite this yet.

References (0)

No backward citations on record.