Rothia species gluten-degrading enzymes and uses thereof

Patent metadata

Field	Value
Publication number	US-9598684-B2
Application number	US-201113808685-A
Country	US
Kind code	B2
Filing date	Jul 7, 2011
Priority date	Jul 7, 2010
Publication date	Mar 21, 2017
Grant date	Mar 21, 2017

Abstract

There are gluten-degrading enzymes found in Rothia species bacteria that are capable of breaking peptide bonds in -XPQ-, -QQP-, -PPF-, -LYP- and/or -PFP-containing peptides. These enzymes are acidic enzymes; they retain protease activity at acidic pH, eg., at 3.0, and have isoelectric points in a pH range of 2.0-7.0. Embodiments herein provide isolated enzyme compositions and probiotic compositions comprising at least one acidic gluten-degrading enzyme or a Rothia species bacteria. Also provided herein are methods of treatment of celiac disease or a related disorder, treatment of gluten-containing foodstuff, degrading and/or detoxifying gluten comprising the acidic gluten-degrading enzyme and/or compositions.

First claim

The invention claimed is:

1. A lyophilized probiotic composition comprising an effective amount of Rothia species bacteria and a pharmaceutically acceptable carrier, wherein the bacteria comprises a gluten-degrading enzyme that retains protease activity at acidic pH of 3.0 as measured in an in vitro gliadin degradation assay for 3 hours using a synthetic substrate Z-YPQ-pNA, wherein the enzyme, when purified, comprises an iso-electric point within a pH range of 2.0-7.0 inclusive, and wherein the composition is formulated into granules.

2. The lyophilized composition of claim 1 , wherein the Rothia species is Rothia species of 188, also known as Rothia sp. HOT-188 and Rothia aeria (strain WSA-8).

3. The lyophilized composition of claim 2 , wherein the iso-electric point of the purified enzyme of the Rothia species bacteria is within a pH range of 2.0-4.0 inclusive.

4. The lyophilized composition of claim 3 , wherein the enzyme comprises a molecular weight between 120-150 kDa inclusive.

5. The lyophilized composition of claim 1 , wherein the pH in which the enzyme retains activity can vary between 2.5 and 5.0, inclusive.

6. The lyophilized composition of claim 1 , wherein the enzyme degrades a gliadin protein, a fragment thereof, or a gluten-containing foodstuff or ingredient thereof.

7. The lyophilized composition of claim 6 , wherein the degradation is partial.

8. The lyophilized composition of claim 6 , wherein the fragment thereof is a 33-mer peptide of α2-gliadin or a 26-mer domain derived from γ-gliadin.

9. The lyophilized composition of claim 8 , wherein the 33-mer peptide is SEQ. ID. NO: 1.

10. The lyophilized composition of claim 8 , wherein the 26-mer domain is SEQ. ID. NO: 2.

11. The lyophilized composition of claim 6 , wherein degradation occurs by cleaving the peptide bond after an amino acid sequence selected from the group consisting of -XPQ- -QQP-, -PPF-, -LYP- and/or -PFP-.

12. The lyophilized composition of claim 1 , wherein the lyophilized composition is admixed with a gluten-containing foodstuff.

13. The lyophilized composition of claim 1 , wherein the granules are packaged into a capsule.

14. The lyophilized composition of claim 1 , wherein the composition is formulated to comprise an enteric coating.

Assignees

• Helmerhorst Eva J
• Oppenheim Frank G
• Univ Boston

Inventors

• Helmerhorst Eva J
• Oppenheim Frank G

Classifications

• A61K38/482

  Serine endopeptidases (3.4.21) · CPC title

• A61P37/08

  Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14) · CPC title

• A23L33/135

  Bacteria or derivatives thereof, e.g. probiotics · CPC title

• C12N9/52 Primary

  derived from bacteria {or Archaea} · CPC title

• A23L33/195

  Proteins from microorganisms · CPC title