Heme-containing cell culture media and uses thereof

Patent metadata

Field	Value
Publication number	US-2025034603-A1
Application number	US-202418915934-A
Country	US
Kind code	A1
Filing date	Oct 15, 2024
Priority date	Oct 20, 2020
Publication date	Jan 30, 2025
Grant date	—

Abstract

The present disclosure provides culture or fermentation media including a biomass or derivative thereof of a hemoprotein-producing C 1 metabolizing non-photosynthetic bacterium, methods of culturing cells or tissue with the culture or fermentation medium, and products produced by the culturing methods including food products, food ingredients, phytoprotective bacterial cell products, and other products of interest such as vitamins, fatty acids, amino acid, carotenoids, etc.

First claim

That which is claimed is:

1 . A method of cell or tissue culture or fermentation comprising culturing cells or tissue in a culture or fermentation medium that comprises a biomass or derivative thereof of a hemoprotein-producing C 1 metabolizing non-photosynthetic bacterium,
wherein the amount of heme in the biomass or derivative thereof is in the range of from 0.01 to 10 mg/g protein of the biomass or derivative thereof, and/or wherein the amount of the biomass or derivative thereof in the culture or fermentation medium is in the range of 0.1 to 50 g/l.

2 . The method of claim 1 , wherein the cells or tissue comprises bacterial cells.

3 . The method of claim 2 , wherein the bacterial cells comprise Bacillus subtilis, Bacillus licheniformis, Escherichia coli, Lactobacillus reuteri, Corynebacterium glutamicum, Pseudomonas putida or Xanthomonas sp.

4 . The method of claim 2 , wherein the bacterial cells comprise Bifidobacterium lactis, Bf. bifidum, Bf. longum, Bf. infantis, Bf. animalis, Bf. breve, Saccharomyces boulardii, Streptococcus Thermophilus, or Bacillus Coagulans.

5 . The method of claim 1 , wherein the bacterial cells comprise marine dwelling bacterial cells.

6 . The method of claim 1 , wherein the cells or tissue comprises non-bacterial cells or tissue.

7 . The method of claim 1 , wherein the cells or tissue comprise algal cells, and optionally the culture or fermentation medium further comprises seawater base.

8 . The method of claim 7 , wherein the algal cells comprise microalgae.

9 . The method of claim 7 , wherein the algal cells comprise macroalgae.

10 . The method of claim 1 , wherein the cells or tissue comprise fungal cells or tissue.

11 . The method of claim 10 , wherein the fungal cells or tissue comprise yeast cells.

12 . The method of claim 10 , wherein the fungal cells or tissue comprise mushroom cells or tissue, and optionally the culture or fermentation medium further comprises potato extract, grain, and/or fruiting substrate.

13 . The method of claim 1 , wherein the cells or tissue comprise animal cells or tissue, and optionally the culture or fermentation medium further comprises serum, a growth hormone, a growth factor, an antibacterial agent and/or an antifungal agent.

14 . The method of claim 12 , wherein the animal cells or tissue comprise fish or shellfish cells or tissue, and optionally the culture or fermentation medium further comprises fish serum.

15 . The method of claim 12 , wherein the animal cells or tissue comprise avian cells or tissue.

16 . The method of claim 1 , wherein the animal cells comprise insect cells or tissue.

17 . The method of claim 12 , wherein the animal cells or tissue comprise mammalian cells or tissue.

18 . The method of claim 17 , wherein the animal cells comprise embryonic stem cells, adult stem cells, myosatellite cells, myoblasts, myocytes, and/or muscle cells.

19 . The method of any one of claims 1 to 18 , wherein the C 1 metabolizing non-photosynthetic bacterium comprises a methylotrophic bacterium.

20 . The method of any one of claims 1 to 18 , wherein the C 1 metabolizing non-photosynthetic bacterium comprises a methanotrophic bacterium.

21 . The method of any claim 18 , wherein the methanotrophic bacterium is selected from the group consisting of Methylomonas, Methylobacter, Methylococcus, Methylosinus, Methylocystis, Methylomicrobium, Methanomonas, and Methylocella.

22 . The method of any of claims 20 , wherein the methanotrophic bacterium comprises Methylococcus capsulatus.

23 . The method of claim 20 , wherein the methanotrophic bacterium comprises Methylococcus capsulatus (Bath).

24 . The method of any one of claims 1 to 23 , wherein the biomass of the hemoprotein-producing C 1 metabolizing non-photosynthetic bacterium or derivative thereof comprises a suspension, an isolate, an autolysate, an homogenate, a digestate, a hydrolysate, an isolate, an extract, or a combination thereof of the biomass.

25 . The method of any one of claims 1 to 24 , wherein the biomass or derivative thereof comprises essential amino acids at an amount within the range of 1-100 mg/g each.

26 . The method of any one of claims 1 to 25 , wherein the biomass or derivative thereof comprises copper at an amount within the range of 50-500 mg/kg.

27 . The method of any one of claims 1 to 26 , wherein the biomass or derivative thereof comprises iron at an amount within the range of 0.05 to 0.6 mg/g.

28 . The method of any one of claims 1 to 27 , further comprising separating the cultured cells from the growth medium to produce isolated culture cells and/or an isolated supernatant.

29 . The method of any of one of claims 1 to 28 , further comprising isolating, concentrating, separating or purifying a desired product from the fermented or cultured cells or tissue.

30 . The method of claim 29 , wherein the desired product is selected from vitamins, fatty acids, amino acids, nucleosides, peptides, proteins, enzymes, pigments, flavors, fragrances, organic acids, preservatives, small molecule metabolites, ferment, culture, probiotics, carotenoids, and meat alternatives.

31 . The method of claim 29 , wherein the desired product is a small molecule metabolite selected from citric acid and lactic acid.

32 . The method of claim 29 , wherein the desired product is an enzyme selected from chymosin, protease, lipase, amylase, cellulase, amylase, and carbohydrase.

33 . The method of claim 29 , wherein the desired product is an omega-3 fatty acid selected from eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA).

34 . The method of claim 29 , wherein the desired product comprises a biomass of cultured cells or tissue.

35 . The method of claim 34 , wherein the biomass of the cultured cells or tissue comprises a probiotic, a dairy culture, or a meat-curing culture.

36 . The method of any of claims 1 to 35 , wherein the biomass of the cultured cells or tissue comprises an elevated level of iron, heme or hemoprotein as compared to a biomass of cells or tissue cultured in a reference culture medium that does not comprise a biomass or derivative thereof of a hemoprotein-producing C 1 metabolizing non-photosynthetic bacterium.

37 . The method of claim 36 , wherein the biomass or derivative thereof of the hemoprotein-producing C 1 metabolizing non-photosynthetic bacterium comprises an autolysate of the hemoprotein-producing C 1 metabolizing non-photosynthetic bacterium.

38 . The method of claim 37 , wherein the cultured cells or tissue, biomass thereof, or derivative of the biomass is suitable for preparing a cell-based meat product or a meat alternative product.

39 . The method of any of claims 1 to 38 , wherein the culturing results in an improved growth rate, yield or productivity of the cells or tissue or the final target ingredient of the culture, as compared to culturing the cells or tissue in a reference culture medium that does not comprise a biomass or derivative thereof of a hemoprotein-producing C 1 metabolizing non-photosynthetic bacterium.

40 . A cell or tissue culture or fermentation m

Assignees

• Calysta Inc

Inventors

• Schiff-Deb Celine
• Giver Lorraine Joan
• Saville Renee Michelle
• Newman Lisa Marie
• Kwan Warren
• Hwang Chien-Seng
• Luning Eric

Classifications

• C12N2511/00

  Cells for large scale production · CPC title

• C12N5/0601

  Invertebrate cells or tissues, e.g. insect cells; Culture media therefor · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N1/16

  Yeasts; Culture media therefor · CPC title

• C12N1/14

  Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor · CPC title