Process for enzymatic hydrolysis of lignocellulosic material and fermentation of sugars

• US 10,717,995 B2
• Filed: 10/04/2018
• Issued: 07/21/2020
• Est. Priority Date: 11/09/2012
• Status: Active Grant

• Current Assignee: Versalis S.P.A, (Eni Petroleum Company Incorporated)
• Original Assignee: DSM IP Assets BV (DSM-Firmenich AG)
• Inventors: Noordam, Bertus, Berkhout, Michael Petrus Jozef
• Primary Examiner(s): Harward, Soren
• Assistant Examiner(s): Pyla, Paul D.
• Application Number:
  US16/152,197
• Publication Number:
  US 20190032093A1
• Time in Patent Office: 656 Days
• Field of Search: None
• CPC Class Codes: C12P 17/10 Nitrogen as only ring hetero atom
• CPC Class Codes: C12P 19/02 Monosaccharides 2-ketogulonic acid C12P7/60
• CPC Class Codes: C12P 19/14 produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase , e.g. by cellulase, hemicellulase
• CPC Class Codes: C12P 2203/00 Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source ethanol C12P7/10
• CPC Class Codes: C12P 35/00 Preparation of compounds having a 5-thia-1-azabicyclo [4.2.0] octane ring system, e.g. cephalosporin
• CPC Class Codes: C12P 5/026 Unsaturated compounds, i.e. alkenes, alkynes or allenes
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• CPC Class Codes: C12P 7/16 Butanols
• CPC Class Codes: C12P 7/18 polyhydric
• CPC Class Codes: C12P 7/20 Glycerol
• CPC Class Codes: C12P 7/40 containing a carboxyl group including Peroxycarboxylic acids
• CPC Class Codes: C12P 7/42 Hydroxy-carboxylic acids
• CPC Class Codes: C12P 7/46 Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
• CPC Class Codes: C12P 7/48 Tricarboxylic acids, e.g. citric acid
• CPC Class Codes: C12P 7/54 Acetic acid vinegar C12J
• CPC Class Codes: C12P 7/56 Lactic acid
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel
• CPC Class Codes: Y02E 50/30 Fuel from waste, e.g. synthetic alcohol or diesel

Abstract

The invention relates to a process for the preparation of a fermentation product from ligno-cellulosic material, comprising the following steps:

• a) optionally pre-treatment of the ligno-cellulosic material;
• b) optionally washing of the optionally pre-treated ligno-cellulosic material;
• c) enzymatic hydrolysis of the optionally washed and/or optionally pre-treated ligno-cellulosic material using an enzyme composition comprising at least two cellulase and whereby the enzyme composition at least comprises GH61;
• d) whereby less than 7.5 mg enzyme composition/g glucan (on dry matter and enzyme as protein) or less than 3.0 mg enzyme composition/g feedstock (on dry matter and enzyme as protein) is used; and
• e) fermentation of the hydrolysed ligno-cellulosic material to produce a fermentation product; and
• f) optionally recovery of a fermentation product;
  wherein before and/or during the enzymatic hydrolysis oxygen is added to the ligno-cellulosic material.

Claims

1. A process for preparation of a sugar product from ligno-cellulosic material, comprising:

enzymatic hydrolysis of ligno-cellulosic material using an enzyme composition comprising at least two cellulases and whereby the enzyme composition at least comprises GH61;

whereby between 0.1 and 7.5 mg enzyme composition/g glucan (on dry matter and enzyme as protein) or between 0.05 and 3.0 mg enzyme composition/g feedstock (on dry matter and enzyme as protein) is used; and

wherein;

the enzymatic hydrolysis occurs for a first part of time and a subsequent second part of time;

during the first part of time dissolved oxygen in the ligno-cellulosic material is at a concentration that is at least 50% less than during the second part of time, during the second part of time oxygen is added to the ligno-cellulosic material continuously, and the second part of time is 2% to 80% of the total enzymatic hydrolysis time; and

the ligno-cellulosic material has a dry matter content of 10 wt % or more.

3. The process according to claim 1, wherein the second part of time is 12% to 50% of the total enzymatic hydrolysis time.

4. The process according to claim 1, wherein the enzymatic hydrolysis is performed in a reactor having a volume of 1 m³ or more.

5. The process according to claim 1, wherein the enzyme composition used retains activity for 30 hours or more.

6. The process according to claim 1, wherein the enzymatic hydrolysis is conducted at a temperature of 45°

C. or more.

7. The process according to claim 1, wherein the enzyme composition is from a fungus, or the enzyme composition comprises a fungal enzyme.

8. The process according to claim 1, in which the enzymatic hydrolysis takes place in a batch, fed batch, and/or continuous culture reactor.

9. The process according to claim 1, in which oxygen is introduced as an oxygen-containing gas in the form of bubbles.

17. The process according to claim 1, wherein the second part of time is 20% to 40% of the total enzymatic hydrolysis time.

18. The process according to claim 1, wherein the dissolved oxygen concentration is maintained in the range of 0.02 mol/m³ to 0.045 mol/m³, and wherein dissolved oxygen is measured under normal atmospheric pressure and at about 62°

C.

19. The process according to claim 1, wherein the dry matter content is 14 to 33 wt %.

20. The process according to claim 7, wherein the fungus is of the genus Rasamsonia, or the fungal enzyme is a Rasamsonia enzyme.

2. A process for preparation of a fermentation product from ligno-cellulosic material, comprising:

enzymatic hydrolysis of ligno-cellulosic material using an enzyme composition comprising at least two cellulases and whereby the enzyme composition at least comprises GH61;

whereby between 0.1 and 7.5 mg enzyme composition/g glucan (on dry matter and enzyme as protein) or between 0.05 and 3.0 mg enzyme composition/g feedstock (on dry matter and enzyme as protein) is used; and

wherein;

the enzymatic hydrolysis occurs for a first part of time and a subsequent second part of time;

during the first part of time dissolved oxygen in the ligno-cellulosic material is at a concentration that is at least 50% less than during the second part of time, during the second part of time oxygen is added to the ligno-cellulosic material discontinuously, and the second part of time is 2% to 80% of the total enzymatic hydrolysis time; and

the ligno-cellulosic material has a dry matter content of 10 wt % or more.

10. The process according to claim 2, wherein the second part of time is 12% to 50% of the total enzymatic hydrolysis time.

11. The process according to claim 2, wherein the enzymatic hydrolysis time is 5 to 150 hours.

12. The process according to claim 2, wherein the enzyme composition used retains activity for 30 hours or more.

13. The process according to claim 2, wherein the enzymatic hydrolysis is conducted at a temperature of 45°

C. or more.

14. The process according to claim 2, wherein the enzyme composition is from a fungus, or the enzyme composition comprises a fungal enzyme.

15. The process according to claim 2, in which the enzymatic hydrolysis takes place in a batch, fed batch, and/or continuous culture reactor.

16. The process according to claim 2, in which oxygen is introduced as an oxygen-containing gas, in the form of bubbles.

21. The process according to claim 2, wherein the second part of time is 20% to 40% of the total enzymatic hydrolysis time.

22. The process according to claim 2, wherein the dissolved oxygen concentration is maintained in the range of 0.02 mol/m³ to 0.045 mol/m³, and wherein dissolved oxygen is measured under normal atmospheric pressure and at about 62°

C.

23. The process according to claim 2, wherein the dry matter content is 14 to 33 wt %.

24. The process according to claim 14, wherein the fungus is of the genus Rasamsonia, or the fungal enzyme is a Rasamsonia enzyme.