Ground meat replicas

Patent metadata

Field	Value
Publication number	US-11439166-B2
Application number	US-202117531460-A
Country	US
Kind code	B2
Filing date	Nov 19, 2021
Priority date	Mar 31, 2014
Publication date	Sep 13, 2022
Grant date	Sep 13, 2022

Abstract

This document relates to ground meat replicas, and more particularly to plant-based products that mimic ground meat, including the fibrousness, heterogeneity in texture, beefy flavor, and red-to-brown color transition during cooking of ground meat.

First claim

What is claimed is:

1. A recombinant Pichia pastoris organism comprising:
eight stably integrated genes encoding recombinant heme biosynthesis enzymes; and
a stably integrated nucleic acid encoding a recombinant heme-containing protein selected from leghemoglobin and myoglobin,
wherein the heme biosynthesis enzymes are Pichia pastoris ALA synthase, Pichia pastoris ALA dehydratase, Pichia pastoris porphobilinogen deaminase, Pichia pastoris uroporphyrinogen III synthase, Pichia pastoris uroporphyrinogen III decarboxylase, Pichia pastoris coproporphyrinogen oxidase, Pichia pastoris protoporphyrinogen III oxidase, and Pichia pastoris ferrochelatase.

2. The recombinant Pichia pastoris organism of claim 1 , wherein at least one of the eight genes and/or the recombinant heme-containing protein is expressed by a methanol inducible promoter.

3. The recombinant Pichia pastoris organism of claim 2 , wherein the methanol inducible promoter is pAOXI.

4. The recombinant Pichia pastoris organism of claim 1 , wherein the heme-containing protein is a mammalian myoglobin.

5. The recombinant Pichia pastoris organism of claim 1 , wherein the heme-containing protein is a Glycine max leghemoglobin.

6. A method of producing a consumable heme-containing protein, the method comprising:
fermenting the recombinant Pichia pastoris organism of claim 2 in a fermentation broth, and inducing expression of the heme-containing protein with methanol.

7. The method of claim 6 , comprising centrifuging cells of the Pichia pastoris organism expressing the heme-containing protein.

8. The method of claim 6 , comprising lysing cells of the Pichia pastoris organism expressing the heme-containing protein to produce a cell lysate.

9. The method of claim 8 , comprising filtering the cell lysate to produce a soluble lysate.

10. The method of claim 9 , comprising concentrating the heme-containing protein in the soluble lysate.

11. The method of claim 6 , comprising centrifuging cells of the Pichia pastoris organism expressing the heme-containing protein, lysing cells of the Pichia pastoris organism expressing the heme-containing protein to produce a cell lysate, filtering the cell lysate to produce a soluble lysate, and concentrating the heme-containing protein in the soluble lysate.

12. The method of claim 6 , wherein the heme-containing protein is a mammalian myoglobin.

13. The method of claim 6 , wherein the heme-containing protein is a Glycine max leghemoglobin.

14. A method for preparing a food additive comprising recombinant heme-containing protein, the method comprising:
providing a recombinant Pichia pastoris organism comprising: eight stably integrated genes encoding recombinant heme biosynthesis enzymes; and a stably integrated gene encoding a recombinant heme-containing protein selected from leghemoglobin and myoglobin under the control of a methanol inducible promoter, wherein the recombinant heme biosynthesis enzymes comprise Pichia pastoris ALA synthase, Pichia pastoris ALA dehydratase, Pichia pastoris porphobilinogen deaminase, Pichia pastoris uroporphyrinogen III synthase, Pichia pastoris uroporphyrinogen III decarboxylase, Pichia pastoris coproporphyrinogen oxidase, Pichia pastoris protoporphyrinogen III oxidase, and Pichia pastoris ferrochelatase;
fermenting the recombinant Pichia pastoris organism in a fermentation broth;
inducing expression of the heme-containing protein with methanol;
purifying the recombinant heme-containing protein; and
combining the purified recombinant heme-containing protein with a buffered solution.

Assignees

• Impossible Foods Inc

Inventors

• Varadan Ranjani
• Solomatin Sergey
• Holz-Schietinger Celeste
• Cohn Elysia
• Klapholz-Brown Ariel
• Shiu Jennifer Woan-Yi
• Kale Aniket
• Karr Jessica
• Fraser Rachel

Classifications

• A23L13/43 Primary

  Addition of vegetable fats or oils; Addition of non-meat animal fats or oils; Addition of fatty acids · CPC title

• A23J3/14

  Vegetable proteins · CPC title

• C12N1/165

  Yeast isolates · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · CPC title

• A23J3/04

  Animal proteins · CPC title