Acidophilic fusarium oxysporum strains, methods of their production and methods of their use

• US 9,796,989 B2
• Filed: 07/02/2015
• Issued: 10/24/2017
• Est. Priority Date: 07/03/2014
• Status: Active Grant

• Current Assignee: Fynder Group Incorporated
• Original Assignee: Sustainable Bioproducts Holdings, LLC
• Inventors: Kozubal, Mark A., Macur, Richard E., Inskeep, William P.
• Primary Examiner(s): DAVIS, RUTH A
• Application Number:
  US14/790,948
• Publication Number:
  US 20160002680A1
• Time in Patent Office: 845 Days
• 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: C12N 1/22 Processes using, or culture media containing, cellulose or hydrolysates thereof
• CPC Class Codes: C12P 2203/00 Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
• CPC Class Codes: C12P 3/00 Preparation of elements or inorganic compounds except carbon dioxide recovery of carbon dioxides as by-products C12F3/02
• CPC Class Codes: C12P 7/065 with microorganisms other than yeasts
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• CPC Class Codes: C12P 7/6458 by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12P 7/649 Biodiesel, i.e. fatty acid alkyl esters
• CPC Class Codes: C12R 2001/77 Fusarium
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The present invention provides isolated acidophilic Fusarium oxysporum strains, such as MK7, and their progeny, compositions comprising such strains and their progeny, methods of producing such strains and their progeny, and methods of using such strains and their progeny.

Claims

1. A method of producing an energy-rich substrate, said method comprising contacting a carbon source with isolated Fusarium oxysporum strain MK7 (ATCC Accession Deposit No. PTA-10698) to form a mixture, incubating said mixture at pH 1.0-4.5 for a period of time under aerobic conditions, and obtaining the energy-rich substrate following such contact, wherein the energy-rich substrate is a total lipid fraction comprising at least 95% C16:

0, C18;

0, C18;

1, and C18;

2 triacylglycerides.

2. The method of claim 1, wherein the carbon source is a cellulosic biomass product.

3. The method of claim 1, wherein the carbon source is a lignocellulosic feedstock or lignocellulosic waste.

4. The method of claim 1, wherein the carbon source is a carbohydrate.

5. The method of claim 4, wherein the carbohydrate is selected from the group consisting of glucose, dextrose, maltose, sucrose, monosaccharides, polysaccharides, oligosaccharides, and starch.

6. The method of claim 1, wherein the Fusarium oxysporum strain and the carbon source are incubated in microaerobic conditions.

7. The method of claim 1, comprising a pretreatment step, wherein said pretreatment step is selected from the group consisting of:

reducing the pH of the carbon source and Fusarium oxysporum mixture, adding manganese to the carbon source and Fusarium oxysporum mixture and adding a nutrient to the carbon source and Fusarium oxysporum mixture before the incubation step.

8. The method of claim 1, wherein the pH is 2.5.

9. The method of claim 1, wherein about 44% of the total lipid content comprises oleic acid (18:

1).

10. The method of claim 1, wherein the energy-rich substrate further comprises a total lipid content of about 46% palmitic (16:

0) and stearic (18;

0) acids.

11. The method of claim 1, wherein the C16:

0 triglyceride comprises at least hexadecanoic acid, methyl ester.

12. The method of claim 1, wherein the C16:

0 triglyceride comprises about 29% palmitic acid.

13. The method of claim 1, wherein the total lipid fraction further comprises about 50% C18:

1-3 triglycerides.

14. The method of claim 13, wherein the C18:

1-3 triglycerides comprise a combination of oleic acid, linoleic acid, conjugated linoleic acids, and linolenic acid.

15. The method of claim 1, wherein the energy-rich substrate further comprises at least one fatty acid methyl ester selected from the group consisting of methyl tetradecanoate;

methyl hexadec-9-enoate;

hexadecanoic acid, methyl ester;

oleic acid;

linoleic acid;

conjugated linoleics;

linolenic acid;

methyl 11-octadecenoate;

octadecanoic acid, methyl ester;

eicosanoic acid, methyl ester;

docosanoic acid, methyl ester; and

tetracosanoic acid, methyl ester.

16. The method of claim 15, wherein the energy-rich substrate further comprises at least one fatty acid selected from the group consisting of methyl tetradecanoate;

eicosanoic acid, methyl ester;

docosanoid acid, methyl ester; and

tetracosanoic acid, methyl ester.

17. The method of claim 1, further comprising extracting lipids from the energy-rich substrate.

18. The method of claim 17, wherein the incubation time is at about 10 days and the total yield in grams of the extracted lipids is about 5% of the initial mass in grams of the carbon source.

19. The method of claim 18, wherein the total yield in grams of the extracted lipids is about 12% of the initial mass in grams of the carbon source.

20. The method of claim 1, wherein the total lipid fraction produced is about 0.05 g/g feedstock.