Treatment of gluten intolerance and related conditions
Provided herein are compositions, foods comprising nepenthesin or a derivative thereof and methods of using nepenthesin or a derivative thereof for modulating gluten intolerance and related conditions, such as celiac disease. Further provided herein are pharmaceutical compositions comprising nepenthesin or a derivative thereof and methods of using nepenthesin or a derivative thereof to treat bacterial infections of the gastrointestinal tract, such as C. difficile or H. pylori . Further provided herein are compositions comprising recombinant nepenthesin I or nepenthesin II, or homologous proteins, and methods for making the same.
1. A composition comprising a recombinant nepenthesin and a transgenic organism wherein said transgenic organisms expresses said nepenthesin and comprises an expression vector, said expression vector comprising a nucleic acid sequence encoding said nepenthesin, wherein said nepenthesin encoded by said nucleic acid sequence comprises an amino acid sequence that is at least 95% identical to the amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 without a plant signal sequence and having aspartic protease activity, wherein said organism is Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, Lactobacillus , or Aspergilli.
2. The composition of claim 1 , wherein said recombinant nepenthesin comprises the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.
3. The composition of claim 1 , wherein said recombinant nepenthesin comprises the amino acid sequence of SEQ ID NO: 6.
4. The composition of claim 1 , wherein said recombinant nepenthesin comprises the amino acid sequence of SEQ ID NO: 7.
5. The composition of claim 1 , wherein said transgenic organism is Escherichia coli.
6. The composition of claim 1 , wherein said nucleic acid sequence encoding said nepenthesin has at least 95% sequence identity with a cDNA that encodes the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.
7. The composition of claim 1 , wherein said nucleic acid sequence encoding said nepenthesin has at least 95% sequence identity with a cDNA that encodes the amino acid sequence of SEQ ID NO: 6 or SEQ ID NO: 7.
8. An expression vector comprising a nucleic acid sequence encoding a nepenthesin, wherein said nepenthesin encoded by said nucleic acid sequence comprises an amino acid sequence that is at least 95% identical to the amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 without a plant signal sequence and having aspartic protease activity.
9. The expression vector of claim 8 , wherein said nepenthesin comprises the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.
10. The expression vector of claim 8 , wherein said nepenthesin comprises the amino acid sequence of SEQ ID NO: 6.
11. The expression vector of claim 8 , wherein said nepenthesin comprises the amino acid sequence of SEQ ID NO: 7.