US2006234320A1PendingUtilityA1

Pyroglutamyl peptidase and its gene

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Assignee: MACHIDA MASAYUKIPriority: Dec 27, 2001Filed: Dec 26, 2002Published: Oct 19, 2006
Est. expiryDec 27, 2021(expired)· nominal 20-yr term from priority
Inventors:Masayuki MachidaKeietsu AbeKatsuya GomiKiyoshi AsaiMotoaki SanoTaishin KinHideki NagasakiAkira HosoyamaOsamu AkitaNaotake OgasawaraSatoru KuharaChikara TokunagaItaru TodaChiaki SaitohAkihiro Senoh
C12P 21/06C12N 9/80C12N 9/0022C12N 9/48
48
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Claims

Abstract

The present invention provides DNA encoding novel pyroglutamyl peptidase derived from Aspergillus oryzae, pyroglutamyl peptidase which is produced by using the DNA, and a method for producing a protein lysate with a good flavor at a high hydrolysis rate.

Claims

exact text as granted — not AI-modified
1 . A polypeptide selected from the group consisting of the following (a) to (c): 
 (a) a polypeptide comprising an amino acid sequence shown in SEQ ID NO: 2,    (b) a polypeptide comprising an amino acid sequence substantially identical to the amino acid sequence shown in SEQ ID NO: 2, and having a pyroglutamyl peptidase activity, and    (c) a polypeptide comprising an amino acid sequence wherein one or more amino acid residues are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 2, and having a pyroglutamyl peptidase activity.    
     
     
         2 . DNA comprising a nucleotide sequence encoding the polypeptide according to  claim 1 .  
     
     
         3 . DNA selected from the group consisting of the following (a) to (c): 
 (a) DNA comprising a nucleotide sequence shown in SEQ ID NO: 1,    (b) DNA comprising a nucleotide sequence shown in SEQ ID NO: 5, and    (c) DNA which hybridizes with DNA comprising a nucleotide sequence complementary to the nucleotide sequence shown in SEQ ID NO: 1 or 5 under stringent conditions, and comprises a nucleotide sequence encoding a polypeptide having a pyroglutamyl peptidase activity.    
     
     
         4 . DNA selected from the group consisting of the following (a) to (c): 
 (a) DNA comprising a nucleotide sequence shown in SEQ ID NO: 3,    (b) DNA which comprises a partial sequence of the nucleotide sequence shown in SEQ ID NO: 3 consisting of 100 or more nucleotides and functions as a promoter, and    (c) DNA which hybridizes with DNA comprising a nucleotide sequence complementary to the nucleotide sequence shown in SEQ ID NO: 3 under stringent conditions and functions as a promoter.    
     
     
         5 . DNA selected from the group consisting of the following (a) to (c): 
 (a) DNA comprising a nucleotide sequence complementary to a nucleotide sequence shown in SEQ ID NO: 4,    (b) DNA which comprises a partial sequence of a nucleotide sequence complementary to the nucleotide sequence shown in SEQ ID NO: 4 consisting of 15 or more nucleotides, and    (c) DNA which hybridizes with DNA comprising the nucleotide sequence shown in SEQ ID NO: 4 under stringent conditions.    
     
     
         6 . The DNA according to any one of  claims 2  to  5 , wherein DNA is genomic DNA.  
     
     
         7 . An oligonucleotide comprising a nucleotide sequence consisting of 15 or more contiguous nucleotides of the nucleotide sequence of the DNA according to any one of  claims 2  to  5  or a nucleotide sequence complementary thereto.  
     
     
         8 . A recombinant DNA comprising the DNA according to claims  2  or  3 .  
     
     
         9 . A transformant comprising the recombinant DNA according to  claim 8 .  
     
     
         10 . A process for producing the polypeptide according to  claim 1 , which comprises culturing a microorganism having an ability to produce the polypeptide in a medium, so as to produce and accumulate the polypeptide in a culture, and recovering the polypeptide from the culture.  
     
     
         11 . The process according to  claim 10 , wherein the microorganism is a transformant comprising a recombinant DNA comprising DNA comprising a nucleotide sequence encoding a polypeptide selected from the group consisting of the following (a) to (c): 
 (a) a polypeptide comprising an amino acid sequence shown in SEQ ID NO: 2,    (b) a polypeptide comprising an amino acid sequence substantially identical to the amino acid sequence shown in SEQ ID NO: 2, and having a pyroglutamyl peptidase activity, and    (c) a polypeptide comprising an amino acid sequence wherein one or more amino acid residues are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 2, and having a pyroglutamyl peptidase activity.    
     
     
         12 . The process according to  claim 10 , wherein the microorganism is filamentous fungus.  
     
     
         13 . The process according to  claim 12 , wherein the filamentous fungus belongs to one genus selected from a group consisting of  Aspergillus, Penicillium, Humicola, Trichoderma, Mucor,  and  Fusarium.    
     
     
         14 . The process according to  claim 13 , wherein the filamentous fungus belonging to  Aspergillus  belongs to one species selected from a group consisting of  Aspergillus oryzae, Aspergillus sojae, Aspergillus niger, Aspergillus awamori, Aspergillus kawachii, Aspergillus parasiticus, Aspergillus flavus, Aspergillus nomius, Aspergillus fumigatus,  and  Aspergillus nidulans.    
     
     
         15 . A process for producing a protein hydrolysate, which comprises adding the polypeptide according to  claim 1  and a protein hydrolase to a raw material containing a protein, and degrading the protein.  
     
     
         16 . A process for producing a protein hydrolysate, which comprises adding a culture containing the polypeptide according to  claim 1  which is obtained by culturing a microorganism having an ability to produce the polypeptide according to  claim 1  in a medium, or a treated product thereof, and a protein hydrolase, to a raw material containing a protein, and degrading the protein.  
     
     
         17 . The process according to  claim 16 , wherein the microorganism is a transformant comprising a recombinant DNA comprising DNA comprising a nucleotide sequence encoding a polypeptide selected from the group consisting of the following (a) to (c): 
 (a) a polypeptide comprising an amino acid sequence shown in SEQ ID NO: 2,    (b) a polypeptide comprising an amino acid sequence substantially identical to the amino acid sequence shown in SEQ ID NO: 2, and having a pyroglutamyl peptidase activity, and    (c) a polypeptide comprising an amino acid sequence wherein one or more amino acid residues are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 2, and having a pyroglutamyl peptidase activity.    
     
     
         18 . The process according to  claim 16 , wherein the microorganism is filamentous fungus.  
     
     
         19 . The process according to  claim 18 , wherein the filamentous fungus belongs to one genus selected from a group consisting of  Aspergillus, Penicillium, Humicola, Trichoderma, Mucor,  and  Fusarium.    
     
     
         20 . The process according to  claim 19 , wherein the filamentous fungus belonging to  Aspergillus  belongs to one species selected from a group consisting of  Aspergillus oryzae, Aspergillus sojae, Aspergillus niger, Aspergillus awamori, Aspergillus kawachii, Aspergillus parasiticus, Aspergillus flavus, Aspergillus nomius, Aspergillus fumigatus,  and  Aspergillus nidulans.    
     
     
         21 . A protein hydrolysate which is produced by the process according to any one of  claims 15  to  20 .  
     
     
         22 . An antibody which specifically binds to the polypeptide according to  claim 1 .  
     
     
         23 . A method of detecting or quantifying the polypeptide according to  claim 1  which comprises using the antibody according to  claim 22 .  
     
     
         24 . An oligonucleotide comprising a nucleotide sequence consisting of 15 or more contiguous nucleotides of the nucleotide sequence of the DNA according to  claim 6  or a nucleotide sequence complementary thereto.  
     
     
         25 . The process according to  claim 10 , wherein the microorganism is a transformant comprising a recombinant DNA comprising DNA selected from the group consisting of the following (a) to (c): 
 (a) DNA comprising a nucleotide sequence shown in SEQ ID NO: 1,    (b) DNA comprising a nucleotide sequence shown in SEQ ID NO: 5, and    (c) DNA which hybridizes with DNA comprising a nucleotide sequence complementary to the nucleotide sequence shown in SEQ ID NO: 1 or 5 under stringent conditions, and comprises a nucleotide sequence encoding a polypeptide having a pyroglutamyl peptidase activity.    
     
     
         26 . The process according to  claim 16 , wherein the microorganism is a transformant comprising a recombinant DNA comprising DNA selected from the group consisting of the following (a) to (c): 
 (a) DNA comprising a nucleotide sequence shown in SEQ ID NO: 1,    (b) DNA comprising a nucleotide sequence shown in SEQ ID NO: 5, and    (c) DNA which hybridizes with DNA comprising a nucleotide sequence complementary to the nucleotide sequence shown in SEQ ID NO: 1 or 5 under stringent conditions, and comprises a nucleotide sequence encoding a polypeptide having a pyroglutamyl peptidase activity.    
     
     
         27 . A protein hydrolysate which is produced by the process of  claim 26.

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