PROCESS OF EXTRACTING OIL FROM THIN STILLAGE

• US 20160152922A1
• Filed: 06/20/2014
• Published: 06/02/2016
• Est. Priority Date: 06/24/2013
• Status: Active Grant

• Current Assignee: Novozymes A/S (Novozymes)
• Original Assignee: Novozymes A/S (Novozymes)
• Inventors: Kreel, Nathaniel, Jump, Joseph
• Granted Patent: US 10,035,973 B2
• Time in Patent Office: Days
• CPC Class Codes: B01D 3/002 by continuous methods
• CPC Class Codes: C11B 13/00 Recovery of fats, fatty oils or fatty acids from waste materials
• CPC Class Codes: C11B 3/003 by enzymes or microorganisms, living or dead
• CPC Class Codes: C12N 9/2417 from microbiological source
• CPC Class Codes: C12N 9/2428 Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
• CPC Class Codes: C12N 9/50 Proteinases , e.g. Endopeptidases (3.4.21-3.4.25)
• CPC Class Codes: C12N 9/52 derived from bacteria or Archaea
• CPC Class Codes: C12N 9/58 derived from fungi
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• CPC Class Codes: C12P 7/64 Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
• CPC Class Codes: C12Y 302/01001 Alpha-amylase (3.2.1.1)
• CPC Class Codes: C12Y 302/01003 Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
• CPC Class Codes: C12Y 304/00 Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel
• CPC Class Codes: Y02W 30/74 Recovery of fats, fatty oils, fatty acids or other fatty substances, e.g. lanolin or waxes

Abstract

The present invention relates to processes of recovering oil after liquefaction and/or from thin stillage and/or syrup/evaporated centrate from a fermentation product production process by adding a thermostable protease to the whole stillage, thin stillage and/or syrup.

Claims

1. A process of recovering oil, comprising

(a) converting a starch-containing material into dextrins with an alpha-amylase;

optionally recovering oil during and/or after step (a)

(b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar;

(c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism;

(d) recovering the fermentation product to form a whole stillage;

(e) separating the whole stillage into thin stillage and wet cake;

(e′

) optionally concentrating the thin stillage into syrup;

(f) recovering oil from the thin stillage and/or optionally the syrup, wherein a protease having a thermostability value of more than 20% determined as Relative Activity at 80°

C./70°

C. is present and/or added during step (a) or steps (d)-(e′

).

2. The process of claim 1, wherein the protease is present in and/or added in starch-containing material converting step (a).

3. The process of claim 1, wherein the temperature in step (a) is above the initial gelatinization temperature, such as at a temperature between 80-90°

C., such as around 85°

C.

4. The process of recovering oil of claim 1, comprising

(a) converting a starch-containing material into dextrins with an alpha-amylase at a temperature above the initial gelatinization temperature;

(b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar;

(c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism;

(d) recovering the fermentation product to form a whole stillage;

(e) separating the whole stillage into thin stillage and wet cake;

(e′

) optionally concentrating the thin stillage into syrup;

(f) recovering oil from the thin stillage and/or optionally the syrup, wherein a protease having a thermostability value of more than 20% determined as Relative Activity at 80°

C./70°

C. is present and/or added during step (a) or any of claims (d)-(e′

).

5. The process of recovering oil of claim 1, comprising

(a) converting a starch-containing material into dextrins with an alpha-amylase at a temperature below the initial gelatinization temperature;

(b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar;

(c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism;

(d) recovering the fermentation product to form a whole stillage;

(e) separating the whole stillage into thin stillage and wet cake;

(e′

) optionally concentrating the thin stillage into syrup;

(f) recovering oil from the thin stillage and/or optionally the syrup, wherein a protease having a thermostability value of more than 20% determined as Relative Activity at 80°

C./70°

C. is present and/or added during any of step (d)-(e′

).

6. The process of claim 1, wherein the protease has a thermostability of more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90% more than 100%, such as more that 105%, such as more than 110%, such as more than 115%, such as more than 120% determined as Relative Activity at 80°

C./70°

C.

7. The process of claim 1, wherein the protease has a thermostability between 50 and 115%, such as between 50 and 70%, such as between 50 and 60%, such as between 100 and 120%, such as between 105 and 115% determined as Relative Activity at 80°

C./70°

C.

8. The process of claim 1, wherein the protease has a thermostability of more than 12%, more than 14%, more than 16%, more than 18%, more than 20%, more than 25%, more than 30%, more than 40%, more that 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 100%, more than 110% determined as Relative Activity at 85°

C./70°

C.

9. The process of claim 1, wherein the protease is derived from Rhizomucor, such as Rhizomucor miehei, such as the protease shown in SEQ ID NO:

9 or one having a sequence identity thereto of at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%.

10. The process of claim 1, wherein the protease is derived from a strain of Pyrococcus, preferably a strain of Pyrococcus furiosus.

11. The process of claim 1, wherein the protease is the one shown SEQ ID NO:

4 herein, or wherein the protease has at least 60%, such as at least 70%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% identity SEQ ID NO;

4 herein.

12. The process of claim 1, wherein the protease is a bacterial serine protease, such as derived from a strain of Thermobifida, such as Thermobifida fusca, such as the protease shown in SEQ ID NO:

10 or one having a sequence identity thereto of at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%.

13. The process of claim 1, wherein the bacterial alpha-amylase is derived from the genus Bacillus, such as a strain of Bacillus stearothermophilus, in particular a variant of a Bacillus stearothermophilus alpha-amylase, such as the one shown in SEQ ID NO:

3 in WO 99/019467 or SEQ ID NO;

1 herein, in particular the Bacillus stearothermophilus alpha-amylase is truncated, preferably to have from 485-495 amino acids, such as around 491 amino acids.

14. The process of recovering oil of claim 1, comprising

(a) converting a starch-containing material into dextrins with an alpha-amylase;

recovering oil during and/or after step (a)

(b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar;

(c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism;

wherein a protease having a thermostability value of more than 20% determined as Relative Activity at 80°

C./70°

C. is present and/or added in step (a).

15. The process of recovering oil of claim 1, comprising

(a) converting a starch-containing material into dextrins with an alpha-amylase;

(b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar;

(c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism;

(d) recovering the fermentation product to form a whole stillage;

(e) separating the whole stillage into thin stillage and wet cake;

(e′

) concentrating the thin stillage into syrup;

(f) recovering oil from the syrup, wherein a protease having a thermostability value of more than 20% determined as Relative Activity at 80°

C./70°

C. is present and/or added in step (e′

).

16. The process of claim 15, wherein the protease is derived from a strain of Pyrococcus, preferably a strain of Pyrococcus furiosus, such as the one shown in SEQ ID NO:

4 herein.

17. The process of claim 15, wherein the protease is a bacterial serine protease, such as derived from a strain of Thermobifida, such as Thermobifida fusca, such as the protease shown in SEQ ID NO:

10 or one having a sequence identity thereto of at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%.

18. The process of claim 15, wherein the protease is a variant of the metallo protease derived from a strain of the genus Thermoascus, preferably a strain of Thermoascus aurantiacus, especially Thermoascus aurantiacus CGMCC No. 0670, such as a protease variant of the Thermoascus aurantiacus protease shown in SEQ ID NO:

3 herein with the mutations selected from the group consisting of;

A27K+D79L+Y82F+S87G+D104P+A112P+A126V+D142L;

D79L+Y82F+S87G+A112P+D142L;

Y82F+S87G+S70V+D79L+D104P+A112P+D142L;

Y82F+S87G+D79L+D104P+A112P+A126V+D142L.

19. The process of claim 15, wherein the protease is derived from Rhizomucor, such as Rhizomucor miehei, such as the protease shown in SEQ ID NO:

9 or one having a sequence identity thereto of at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%.

20. (canceled)