Process for enzymatic hydrolysis of lignocellulosic material and fermentation of sugars

• US 10,337,040 B2
• Filed: 04/29/2015
• Issued: 07/02/2019
• Est. Priority Date: 04/30/2014
• 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, Bevers, Loes Elizabeth, Parton, Rudy Francois Maria Jozef, Berkhout, Michael Petrus Jozef
• Primary Examiner(s): Holland, Paul J
• Application Number:
  US15/307,241
• Publication Number:
  US 20170145455A1
• Time in Patent Office: 1,525 Days
• Field of Search: None
• 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: C13K 1/02 obtained by saccharification of cellulosic materials manufacture of fodder A23K10/32
• CPC Class Codes: C13K 13/00 Sugars not otherwise provided for in this class
• CPC Class Codes: C13K 13/002 Xylose
• CPC Class Codes: D21C 3/26 Multistage processes
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

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

• • a) optionally, pretreatment of the lignocellulosic material,
  • b) optionally, washing of the optionally pretreated lignocellulosic material,
  • c) enzymatic hydrolysis of the optionally washed and/or optionally pretreated lignocellulosic material using an enzyme composition comprising at least two cellulases and whereby the enzyme composition at least comprises LPMO, and optionally purifying the hydrolysed lignocellulosic material,
  • d) fermentation of the hydrolysed lignocellulosic material to produce a fermentation product, and
  • e) optionally, recovery of a fermentation product,
    • wherein the amounts of formed hydrolysed oxidation products at the end of the enzymatic hydrolysis by the oxidation by LPMO of the lignocellulosic material containing cellulose and/or cello-oligosaccharides is kept between 3 to 80 g/kg glucan present in the lignocellulosic material by adding a suitable amount of oxygen after the pre-treatment and before and/or during the enzymatic hydrolysis to the lignocellulosic material, preferably the formed hydrolysed oxidation product is gluconic acid, an aldonic acid and/or geminal diol, more preferably the hydrolysed oxidation product is gluconic acid.

Claims

1. A process for the preparation of a sugar product from lignocellulosic material, comprising:

enzymatic hydrolysis and oxidation of the lignocellulosic material in the presence of an enzyme composition comprising at least two cellulases and a lytic polysaccharide monooxygenase (LPMO);

adding 20-10,000 mmol of oxygen per kg glucan to the lignocellulosic material during the enzymatic hydrolysis;

wherein the enzymatic hydrolysis is conducted until at least 70% of available sugar in the lignocellulosic material is released; and

wherein at the end of the enzymatic hydrolysis the amount of gluconic acid formed during oxidation of the lignocellulosic material is kept between 3 to 10 g/kg glucan present in the lignocellulosic material.

5. The process of claim 1, wherein the oxygen is added in the form of bubbles.

6. The process of claim 1, wherein the enzymatic hydrolysis is in a reactor with a volume of 1 m³ or more.

7. The process of claim 1, wherein the enzymatic hydryolysis time is 5 to 150 hours.

8. The process of claim 1, wherein the enzyme composition retains activity for 30 hours or more.

9. The process of claim 1, wherein the hydrolysis is conducted at a temperature of 45°

C. or more.

10. The process of claim 1, wherein the enzyme composition comprises a fungal enzyme.

11. The process of claim 1, wherein the dry matter content in the enzymatic hydrolysis step is 10 wt % or more.

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

13. The process of claim 1, wherein the enzyme composition is in the form of a whole fermentation broth of a fungus.

14. The process of claim 1, wherein oxygen is introduced as air.

15. The process of claim 1, wherein the enzyme composition comprises a Rasamsonia enzyme.

16. The process of claim 9, wherein the enzymatic hydrolysis is conducted at a temperature of 55°

C. or more.

17. The process of claim 11, wherein the dry matter content in the hydrolysis step is 14 to 33 wt %.

19. The process of claim 1, wherein the enzymatic hydrolysis is conducted until at least 80% of available sugar in the lignocellulosic material is released.

20. The process of claim 1, wherein 30-5,000 mmol oxygen per kg glucan is added to the lignocellulosic material during the enzymatic hydrolysis.

21. The process of claim 1, wherein 40-4,000 mmol oxygen per kg glucan is added to the lignocellulosic material during the enzymatic hydrolysis.

22. The process of claim 1, wherein 50-3,500 mmol oxygen per kg glucan is added to the lignocellulosic material during the enzymatic hydrolysis.

2. A process for the preparation of a fermentation product from lignocellulosic material, comprising:

a) enzymatic hydrolysis and oxidation of the lignocellulosic material in the presence of an enzyme composition comprising at least two cellulases and and a lytic polysaccharide monooxygenase (LPMO) to produce a hydrolysed lignocellulosic material;

adding 20-10,000 mmol of oxygen per kg glucan to the lignocellulosic material before and/or during the enzymatic hydrolysis

wherein the enzymatic hydrolysis is conducted until at least 70% of available sugar in the lignocellulosic material is released; and

b) fermentation of the hydrolysed lignocellulosic material to produce a fermentation product,

wherein at the end of the enzymatic hydrolysis the amount of gluconic acid formed during LPMO oxidation of the lignocellulosic material is kept between 3 to 10 g/kg glucan present in the lignocellulosic material.

3. The process of claim 2, wherein the fermentation is conducted with a microorganism that is able to ferment at least one C5 sugar.

4. The process of claim 3, wherein the microorganism is a genetically modified Saccharomyces cerevisiae.

18. The process of claim 2, wherein the enzymatic hydrolysis is conducted until at least 80% of available sugar in the lignocellulosic material is released.

23. The process of claim 2, wherein 30-5,000 mmol oxygen per kg glucan is added to the lignocellulosic material during the enzymatic hydrolysis.

24. The process of claim 2, wherein 40-4,000 mmol oxygen per kg glucan is added to the lignocellulosic material during the enzymatic hydrolysis.

25. The process of claim 2, wherein 50-3,500 mmol oxygen per kg glucan is added to the lignocellulosic material during the enzymatic hydrolysis.