FILAMENTOUS FUNGAL BIOMATS, METHODS OF THEIR PRODUCTION AND METHODS OF THEIR USE

• US 20190040352A1
• Filed: 08/30/2018
• Published: 02/07/2019
• Est. Priority Date: 03/01/2016
• Status: Active Grant

• Current Assignee: Fynder Group Incorporated
• Original Assignee: Sustainable Bioproducts, Inc.
• Inventors: KOZUBAL, Mark A., MACUR, Richard E., AVNIEL, Yuval C.
• Granted Patent: US 10,533,155 B2
• Time in Patent Office: Days
• CPC Class Codes: C02F 3/347 Use of yeasts or fungi C02F3/322 takes precedence
• CPC Class Codes: C12N 1/02 Separating microorganisms from their culture media
• CPC Class Codes: C12N 1/14 Fungi culture of mushrooms A01G18/00; as new plants A01H15/00; Culture media therefor
• CPC Class Codes: C12N 1/145 Fungal isolates
• CPC Class Codes: C12R 2001/645 Fungi ; Processes using fungi
• CPC Class Codes: C12R 2001/66 Aspergillus
• CPC Class Codes: C12R 2001/77 Fusarium
• CPC Class Codes: C12R 2001/785 Mucor
• CPC Class Codes: C12R 2001/80 Penicillium
• CPC Class Codes: C12R 2001/84 Pichia
• CPC Class Codes: C12R 2001/845 Rhizopus
• CPC Class Codes: C12R 2001/885 Trichoderma

Abstract

A novel method of growing fungi is disclosed which uses an engineered artificial media and produces high density filamentous fungi biomats that can be harvested with a minimum of processing and from which fungal products such as antibiotics, proteins, and lipids can be isolated, the method resulting in lowered fungus cultivation costs for energy usage, oxygenation, water usage and waste stream production.

Claims

1. A method of producing a filamentous fungi biomat comprising

(a) inoculating an effective amount of planktonic cells of at least one filamentous fungi into an artificial liquid growth media, wherein the liquid growth media'"'"'s carbon source is selected from the group consisting of starch waste, acid whey, corn steep liquor, potato waste, sugar beet waste, glycerol, dextrose, glucose, maltose, galactose, mannose, corn syrup, vegetable scraps, and cattle feedstock water;

(b) incubating the inoculated growth media in an undisturbed state for a period of time;

(c) producing a filamentous fungi biomat; and

(d) optionally harvesting the filamentous fungi biomat.

2. The method of claim 1 wherein 7.5% (volume:

volume) of planktonic cells are inoculated into the artificial growth media.

3. The method according to claim 1, wherein the carbon source is acid whey.

4. The method according to claim 1, wherein the carbon source is starch.

5. The method according to claim 1, wherein the carbon source is corn steep liquor.

6. The method according to claim 1, wherein the carbon source is dextrose, glucose, maltose, galactose, and/or mannose.

7. The method according to claim 1, wherein the carbon source is corn syrup.

8. The method according to claim 1, wherein the at least one filamentous fungi is selected from the group consisting of Fusarium oxysporum strain MK7 (ATCC Accession Deposit No. PTA-10698), Fusarium species, and Rhizopus species.

9. The method according to claim 8, wherein the at least one filamentous fungi is Fusarium oxysporum strain MK7 (ATCC Accession Deposit No. PTA-10698), Fusarium venenatum and/or Rhizopus oligosporus.

10. The method according to claim 8, wherein the at least one filamentous fungi is Fusarium oxysporum strain MK7 (ATCC Accession Deposit No. PTA-10698).

11. The method according to claim 1, wherein the artificial media has an osmotic pressure of about 18.6 atm.

12. The method according to claim 1, wherein the artificial media has an ionic strength of about 0.368.

13. The method according to claim 1, wherein the liquid media comprises ammonium nitrate, urea, potassium dihydrogen phosphate, calcium chloride, magnesium sulfate, and yeast extract.

14. The method according to claim 13, wherein the liquid media further comprises ferric sulfate, zinc sulfate, manganese chloride, cobalt chloride, copper sulfate, and boric acid.

15. A filamentous fungal biomat comprising filamentous fungal mycelia and/or hyphae and a filamentous fungal cell density of at least 25 g/l (dry wt/l media).

16. The filamentous fungal biomat according to claim 15, wherein the cell density is at least 50 g/l (dry wt/l media) or at least 75 g/l (dry wt/l media).

17. The filamentous fungal biomat of claim 15, wherein the cell density is at least 120 g/l (dry wt/l media).

18. The filamentous fungal biomat of claim 15, wherein the cell density is at least 180 g/l (dry wt/l media).

19. The filamentous fungal biomat of claim 15, wherein the filamentous fungal cells are selected from the group consisting of Fusarium oxysporum strain MK7 (ATCC Accession Deposit No. PTA-10698), Fusarium species, and Rhizopus species.

20. The filamentous biomat according to claim 19, wherein the filamentous fungal cells are Fusarium oxysporum strain MK7 (ATCC Accession Deposit No. PTA-10698), Fusarium venenatum and/or Rhizopus oligosporus.

21. The filamentous biomat according to claim 19, wherein the filamentous fungal cells are Fusarium oxysporum strain MK7 (ATCC Accession Deposit No. PTA-10698).

22. The filamentous fungal biomat according to claim 15, wherein the fungal mycelia and/or hyphae are predominantly organized parallel to the air:

biomat interface and/or biomat;

media interface.

23. The filamentous fungal biomat according to claim 15, wherein the filamentous fungal biomat comprises at least two structurally different cell layers, one cell layer in contact with the artificial media and at least one other cell layer.

24. The filamentous fungal biomat according to claim 23, wherein the structural difference between the cell layers is the density of cells within the layer.

25. The filamentous fungal biomat according to claim 23, wherein one cell layer is in contact with the air and at least one other cell layer.

26. The filamentous fungal biomat according to claim 23, wherein the biomat comprises three structurally different cell layers.

27. The filamentous fungal biomat according to claim 15, wherein the tensile strength of the biomat is at least 0.00315 Kg/mm² (4.48 PSI).

28. The filamentous fungal biomat of claim 15, wherein the biomat has a protein content of at least 40%.

29. The filamentous fungal biomat of claim 15, wherein the biomat has a lipid content of at least 39%.

30. A method of removing carbon and nitrogen from cattle feedlot lagoon water comprising

(a) adjusting the lagoon water to pH 2.6;

(b) inoculating the adjusted lagoon water with strain MK7 (ATCC Accession Deposit No. PTA-10698);

(c) producing a floating filamentous fungi biomat on the surface of the lagoon; and

(d) harvesting the filamentous fungi biomat, wherein the biomat contains the carbon and nitrogen removed from the lagoon water.