Granted Patent B2

US 12,391,935 · App. 18/191,395 · Granted Aug 19, 2025

Inventors: Halle Redfearn (Daly City, CA); Harshal Kshirsagar (Daly City, CA); Kritika Mahadevan (Daly City, CA); Alexandre Chapeaux (Daly City, CA); Wesley Rutherford-Jenkins (Daly City, CA); Joel Andrew Kreps (Daly City, CA); Isha Joshi (Daly City, CA)

Assignee: CLARA FOODS CO.

C12N9/50C12N9/6481C12P21/00C12Y304/23001

Abstract

Claims (19)

1. A method for producing a consumable composition comprising combining a high-activity stable pepsin powdered composition with one or more consumable ingredients comprising steps of:

(a) providing a microorganism expressing a recombinant pepsinogen, wherein the expressed pepsinogen is secreted by the microorganism into its growth media;

(b) harvesting the growth media and removing the microorganismal cells therefrom to obtain a liquid starting material;

(c) lowering the pH of the liquid starting material to less than pH 4.0 to obtain an activated pepsin composition;

(d) raising the activated pepsin composition to a pH of 5.4 or higher to obtain a high-activity stable pepsin composition; and

(e) isolating the stable pepsin polypeptide from protein and small molecules in the liquid starting material to obtain the high-activity stable pepsin powdered composition having a specific activity from 500 FCC units/mg total protein to 50,000 FCC units/mg total protein;

(f) combining the high-activity stable pepsin powdered composition with the one or more consumable ingredients to produce a consumable composition; and

wherein step (e) is performed after steps (c) and (d).

2. The method of claim 1 , wherein the high-activity stable pepsin composition comprises a pepsin polypeptide having an amino acid sequence of a sheep, pig, cow, human, zebu, yak, Central European red deer, or goat pepsin.

3. The method of claim 2 , wherein the recombinant pepsinogen comprises any one of SEQ ID NO: 1 to SEQ ID NO: 9, or an amino acid sequence with at least 90% identity thereto.

4. The method of claim 2 , wherein the high-activity stable pepsin powdered composition comprises at least 5000 FCC units/mg total protein.

5. The method of claim 1 , wherein the microorganism is a Pichia species, a yeast, a filamentous fungus, a Saccharomyces species, a bacterium, a Trichoderma species, or an Aspergillus species.

6. The method of claim 1 , wherein the high-activity stable pepsin powdered composition comprises less than 5% of contaminating proteins.

7. The method of claim 6 , wherein the contaminating protein is a non-pepsin protein.

8. The method of claim 1 , wherein the consumable composition is a food composition.

9. The method of claim 8 , wherein the one or more consumable ingredients comprise a food additive.

10. The method of claim 1 , wherein the consumable composition is a pharmaceutical composition.

11. The method of claim 10 , wherein the one or more consumable ingredients is a pharmaceutically acceptable excipient.

12. The method of claim 1 , further comprising purifying the stable pepsin polypeptide before step (f) wherein after purifying, the high-activity stable pepsin powdered composition comprises less than 5 % of contaminating proteins.

Assignments (1)

Continuity (6)

References Cited (314)