Combination enzyme therapy for digestion of dietary gluten
A combination enzyme product consisting of a glutamine specific endoprotease and a prolyl endopeptidase is provided. Both enzymes are active and stable in the stomach and can therefore be administered as lyophilized powders or simple capsules/tablets. A ratio of the two enzymes is used to maximize their synergy.
1. A method of hydrolyzing gluten proteins to non-toxic metabolites, the method comprising
contacting gluten-containing foodstuff with an enzyme composition comprising: a combination of Sphingomonas capsulata prolyl endopeptidase (SC-PEP) and cysteine endoprotease B, isoform 2 (EP-B2), wherein the ratio of EP-B2:SC-PEP is 1:1 by weight; in a dose of from 100 mg to 2000 mg total enzyme,
wherein the effective dose hydrolyzes gluten proteins to yield non-toxic metabolites.
2. The method of claim 1 , wherein EP-B2 is provided as a proenzyme form of EP-B2.
3. The method of claim 1 , wherein the contacting is performed in vivo.
4. The method of claim 3 , wherein the effective dose is administered in vivo prior to or concurrently with a gluten-containing foodstuff.
5. The method of claim 4 , wherein the effective dose is orally administered.
6. The method of claim 1 , wherein the contacting is performed in vitro.
7. The method of claim 1 , wherein the enzymes are provided separately.
8. The method of claim 1 , wherein the enzymes are provided in a combined formulation.
9. The method of claim 1 , wherein the specific activity of SC-PEP is at least 500 U/mg, where 1 unit is defined as 1 μM p-nitroaniline released per minute from a chromogenic substrate Cbz-Gly-Pro-pNA at room temperature; and the specific activity of EP-B2 is at least 500 U/mg, wherein the specific activity of EP-B2 is defined by the activity of the mature enzyme form, where 1 unit is defined as 1 μM p-nitroaniline released per minute from a chromogenic substrate CBz-Phe-Arg-pNA at room temperature.
10. The method of claim 9 , wherein SC-PEP has a specific activity of at least 1000 U/g, and EP-B2 has a specific activity of at least 1000 U/mg.
11. The method of claim 9 , wherein SC-PEP has a specific activity of about 2500 Units/mg, and proEP-B2 has a specific activity of about 4000 Units/mg.
12. An enzyme composition comprising:
a combination of Sphingomonas capsulata prolyl endopeptidase (SC-PEP) and cysteine endoprotease B, isoform 2 (EP-B2), wherein the ratio of EP-62:SC-PEP is 1:1 by weight,
in a dose of from 100 mg to 2000 mg enzyme.
13. The enzyme combination of claim 12 , wherein EP-B2 is provided as a proenzyme form of EP-B2.
14. The enzyme composition of claim 13 , wherein the enzymes are provided separately.
15. The enzyme composition of claim 13 , wherein the enzymes are provided in a combined formulation.
16. The enzyme composition of claim 13 , wherein the specific activity of SC-PEP is at least 500 U/mg, where 1 unit is defined as 1 μM p-nitroaniline released per minute from a chromogenic substrate Cbz-Gly-Pro-pNA at room temperature; and the specific activity of EP-B2 is at least 500 U/mg, wherein the specific activity of EP-B2 is defined by the activity of the mature enzyme form, where 1 unit is defined as 1 μM p-nitroaniline released per minute from a chromogenic substrate CBz-Phe-Arg-pNA at room temperature.
17. The enzyme combination of claim 16 , wherein SC-PEP has a specific activity of at least 1000 U/g, and EP-B2 has a specific activity of at least 1000 U/mg.
18. The enzyme combination of claim 16 , wherein SC-PEP has a specific activity of about 2500 Units/mg, and proEP-B2 has a specific activity of about 4000 Units/mg.