Use of hydrophobins to increase gas transfer in aerobic fermentation processes

Patent metadata

Field	Value
Publication number	US-9982229-B2
Application number	US-201415102486-A
Country	US
Kind code	B2
Filing date	Nov 14, 2014
Priority date	Dec 19, 2013
Publication date	May 29, 2018
Grant date	May 29, 2018

Abstract

The present disclosure provides methods, compositions and apparatuses for increasing gas transfer in fermentation processes.

First claim

The invention claimed is:

1. A method of increasing gas transfer in an aerobic fermentation process comprising adding one or more Class II hydrophobins to a fermentation medium to cause the increase in gas transfer.

2. The method of claim 1 , where the gas is oxygen.

3. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin having a hydrophobin core between 40 and 120 amino acids.

4. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin fusion protein.

5. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is obtained or obtainable from a filamentous fungus.

6. The method according to claim 5 , wherein at least one of the one or more Class II hydrophobins is obtained or obtainable from a fungus of genus selected from the group consisting of Cladosporium, Ophistoma, Cryphonectria, Trichoderma, Gibberella, Neurospora, Maganaporthe, Hypocrea, Xanthoria, Emericella, Aspergillus, Paracoccioides, Metarhizium, Pleurotus, Coprinus, Dicotyonema, Flammulina, Schizophyllum, Agaricus, Pisolithus, Tricholoma, Pholioka, Talaromyces and Agrocybe.

7. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is generated in situ in the fermentation process.

8. The method of claim 1 , wherein said fermentation process has an equilibrium surface tension at a water/air interface below 70 mN/m, or below 50 mN/m, or below 40 mN/m, or below 30 mN/m.

9. The method of claim 1 , wherein said fermentation process has a surface shear elasticity at a water/air interface of at least 0.5-0.7 N/m.

10. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin that causes at least 1 to 2 fold increase in stability of bubbles in the fermentation medium compared to stability of bubbles in the absence of said hydrophobin.

11. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin that causes at least a 10% increase in the number of bubbles in the fermentation medium compared to the number of bubbles in the fermentation medium in the absence of said hydrophobin.

12. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin that provides a decrease in an equilibrium surface tension of said fermentation medium below 50 mN/m when compared to the equilibrium surface tension of said fermentation in the absence of said hydrophobin.

13. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin that causes a surface shear elasticity of said fermentation medium to increase to 0.5-0.7 N/m or higher when compared to the surface shear elasticity of said fermentation medium in the absence of said hydrophobin.

14. The method of claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin that causes a viscous modulus (G″s) of said fermentation medium to increase to 0.02-0.05 N/m when compared to viscous modulus (G″s) of said fermentation medium in the absence of said hydrophobin.

15. The method according to claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin having the general formula:
(Y1) n -B1-(X1) a -B2-B3-(X3) c -B4-(X4) d -B5-(X5) e -B6-B7-(X7) g -B8-(Y2) m
wherein:
m and n are independently an integer between 0 to 200;
B1, B2, B3, B4, B5, B6, B7 and B8 are each an amino acid selected from the group consisting of Cys, Leu, Ala, Ser, Thr, Met or Gly, wherein at least 6 of the residues B1 through B8 being Cys;
a is an integer between 6 to 12;
c is an integer between 8 to 16;
d is an integer between 2 to 20;
e is an integer between 4 to 12; and
g is an integer between 5 to 15.

16. The method according to claim 1 , wherein at least one of the one or more Class II hydrophobins is a hydrophobin having the general formula:
(Y1) n -B1-(X1) a -B2-B3-(X3) c -B4-(X4) d -B5-(X5) e -B6-B7-(X7) g -B8-(Y2) m
wherein:
m and n are independently an integer between 0 to 10;
B1, B2, B3, B4, B5, B6, B7 and B8 are each an amino acid selected from the group consisting of Cys, Leu or Ser, wherein at least 7 of the residues B1 through B8 being Cys;
a is an integer between 7 to 11;
c is 11;
d is an integer between 4 to 18;
e is an integer between 6 to 10; and
g is an integer between 7 to 10.

17. The method of claim 15 , wherein all 8 of the residues B1 through B8 are Cys.

18. The method of claim 15 , wherein the group (X3)c comprises the sequence motif ZZXZ, wherein Z is an aliphatic amino acid; and X is any amino acid.

19. The method of claim 15 , wherein the hydrophobin is present in a concentration of 0.1-400 μM, or 5-125 μM, or 14-69 μM.

20. The method of claim 15 , wherein the hydrophobin is present in a concentration of 50-1000 mg/kg by weight of the total weight of the fermentation medium.

21. The method of claim 15 , wherein the hydrophobin is present in a concentration of 0.72-2900 mg/L, or 36-900 mg/L, or 100-500 mg/L.

22. The method of claim 1 , wherein said hydrophobin is “HFBII” (SEQ ID NO: 2), or a protein having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, or at least 99% sequence identity in the hydrophobin core thereof.

Assignees

• Danisco Us Inc

Inventors

• Sims Evan
• Schelle Michael W
• Chotani Gopal

Classifications

• Y02E50/17

  Cross-Sectional Technologies · mapped topic

• C07K14/37

  from fungi · CPC title

• C12N1/14

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

• C12N1/12

  Unicellular algae; Culture media therefor (as new plants A01H13/00) · CPC title

• C12N9/00

  Enzymes; Proenzymes; Compositions thereof (preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; {enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113}); Processes for preparing, activating, inhibiting, separating or purifying enzymes (preparation of malt C12C1/00) · CPC title