Proteases for Degrading Gluten
Gluten-degrading proteases derived from insects, including flour beetles, are isolated, and the purified, and recombinant forms can be used to make gluten-containing food safe for patients suffering from gluten intolerance.
1 . A gluten-degrading protease from an insect that feeds on gluten containing dried grain products, in isolated, purified, or recombinant form.
2 . The protease of claim 1 , wherein said insect is a flour beetle.
3 . The protease of claim 1 , wherein said flour beetle is Tribolium castaneum.
4 . The protease of claim 1 wherein said protease is a protease set forth in Table 1, or a homolog, ortholog or variant thereof.
5 . The protease of claim 4 , wherein said homolog, ortholog or variant has at least 80% sequence identity to a protease set forth in Table 1.
6 . The protease of claim 4 , wherein said protease degrades said gluten in a foodstuff to fragments shorter than 8 amino acids.
7 . The protease of claim 4 , wherein said protease digests gluten fragments that are resistant to normal digestive enzymes.
8 . The protease of claim 1 , wherein said protease is formulated with a pharmaceutically acceptable excipient.
9 . The protease of claim 1 , wherein said protease is admixed with food.
10 . The protease according to claim 1 , wherein said protease is stable to acid conditions.
11 . A recombinant expression vector for a gluten-degrading protease, comprising a coding sequence for a protease set forth in claim 1 and a promoter that drives expression of said protease in a suitable host cell,
12 . A method for degrading gluten in food, said method comprising contacting gluten-containing food with one or more proteases of claim 1 .
13 . A pharmaceutical formulation suitable for oral administration that contains a protease of claim 1 admixed with one or more pharmaceutically acceptable excipients.
14 . A pharmaceutical formulation of claim 13 , further comprising one or more non-insect proteases.
15 . The pharmaceutical formulation of claim 14 , wherein said non-insect protease is one or more of Hordeum vulgare endoprotease (Genbank accession U19384); X-Pro dipeptidase from Aspergillus oryzae (GenBank ID#BD191984); carboxypeptidase from Aspergillus saitoi (GenBank ID#D25288); Flavobacterium meningosepticum PEP (Genbank ID #D10980); Sphingomonas capsulata PEP (Genbank ID#AB010298); Penicillium citrinum PEP (Genbank ID#D25535); Lactobacillus helveticus PEP (Genbank ID#321529); and Myxococcus xanthus PEP (Genbank ID#AF127082)
16 . A method for treating gluten intolerance in a patient in need of such treatment, wherein said treatment reduces exposure of said patient to immunogenic gluten peptides, said method comprising the step of orally administering to said patient a therapeutically effective dose of one or more proteases of claim 1 or a pharmaceutical formulation thereof contemporaneously with the ingestion of a food that may contain gluten.
17 . The method of claim 16 , further comprising orally administering to said patient a therapeutically effective dose of one or more non-insect proteases.
18 . The method of claim 17 , wherein said non-insect protease is one or more of Hordeum vulgare endoprotease (Genbank accession U19384); X-Pro dipeptidase from Aspergillus oryzae (GenBank ID#BD191984); carboxypeptidase from Aspergillus saitoi (GenBank ID#D25288); Flavobacterium meningosepticum PEP (Genbank ID #D10980); Sphingomonas capsulata PEP (Genbank ID#AB010298); Penicillium citrinum PEP (Genbank ID#D25535); Lactobacillus helveticus PEP (Genbank ID#321529); and Myxococcus xanthus PEP (Genbank ID#AF127082)