# US 10,865,427 B2 · Process for enzymatic hydrolysis of lignocellulosic material and fermentation of sugars

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- Number: US 10,865,427 B2
- Document ID: US10865427B2
- Status: Granted
- Kind: B2
- Application number: 16/546,651
- Filed: 2019-08-21
- Published: 2019-12-19
- Granted: 2020-12-15
- Earliest claimed: 2014-10-21
- Assignees: Dsm IP Assets B.V.
- Applicants: Dsm IP Assets B.V.
- Inventors: Johannes Petrus Smits; Elisabeth Maria Gierveld
- Web page: https://patentgrep.com/patent/US10865427B2

## CPC

- C12P 7/10: substrate containing cellulosic material
- C02F 3/28: Anaerobic digestion processes
- C02F 3/34: characterised by the microorganisms used
- C12P 7/14: Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms
- C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60)
- C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase}
- C13K 1/02: obtained by saccharification of cellulosic materials (manufacture of fodder A23K10/32)
- C13K 13/00: Sugars not otherwise provided for in this class
- C13K 13/002: {Xylose}
- C12P 2201/00: Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
- C12P 2203/00: Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C02F: TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE (separation in general B01D; special arrangements on waterborne vessels of installations for treating water, waste water or sewage, e.g. for producing fresh water, B63J; adding materials to water to prevent corrosion C23F; treating radioactively-contaminated liquids G21F9/04; regeneration of reactants for recirculation into processes, see the relevant places for the processes)
- C13K: SACCHARIDES OBTAINED FROM NATURAL SOURCES OR BY HYDROLYSIS OF NATURALLY OCCURRING DISACCHARIDES, OLIGOSACCHARIDES OR POLYSACCHARIDES (chemically synthesised sugars or sugar derivatives C07H; polysaccharides, e.g. starch, derivatives thereof C08B; malt C12C; fermentation or enzyme-using processes for preparing compounds containing saccharide radicals C12P19/00; production of sucrose C13B)
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

The invention relates to a process for the preparation of a sugar and/or fermentation product from lignocellulosic material.

## Claims

### Claim 1

A process for preparation of a sugar product from lignocellulosic material, comprising:
a) optionally, pretreatment of the lignocellulosic material;
b) optionally, washing of the optionally pretreated lignocellulosic material;
c) fed-batch addition of the optionally washed and/or optionally pretreated lignocellulosic material to a first container that comprises an enzyme composition comprising at least two cellulases;
d) enzymatic hydrolysis of the optionally washed and/or optionally pretreated lignocellulosic material in the first container using the enzyme composition comprising at least two cellulases to liquefy the lignocellulosic material;
e) addition of the liquefied lignocellulosic material to a second container;
f) enzymatic hydrolysis of the liquefied lignocellulosic material in the second container using an enzyme composition comprising at least two cellulases to obtain a sugar product; and
g) optionally, recovery of the sugar product;
wherein oxygen is added to the first and/or second container.

### Claim 2

A process for preparation of a fermentation product from lignocellulosic material, comprising:
a) optionally, pretreatment of the lignocellulosic material;
b) optionally, washing of the optionally pretreated lignocellulosic material;
c) fed-batch addition of the optionally washed and/or optionally pretreated lignocellulosic material to a first container that comprises an enzyme composition comprising at least two cellulases;
d) enzymatic hydrolysis of the optionally washed and/or optionally pretreated lignocellulosic material in the first container using the enzyme composition comprising at least two cellulases to liquefy the lignocellulosic material;
e) addition of the liquefied lignocellulosic material to a second container;
f) enzymatic hydrolysis of the liquefied lignocellulosic material in the second container using an enzyme composition comprising at least two cellulases to obtain a hydrolysed lignocellulosic material;
g) optionally, recovery of the hydrolysed lignocellulosic material;
h) fermentation of the hydrolysed lignocellulosic material to produce a fermentation product; and
i) optionally, recovery of the fermentation product;
wherein oxygen is added to the first and/or second container.

### Claim 3 (depends on 1)

The process according to claim 1, wherein additional enzymes are added during (d) and/or (f).

### Claim 4 (depends on 1)

The process according to claim 1, wherein the viscosity of the lignocellulosic material in the first container during (d) is controlled by adjusting the addition rate of the optionally washed and/or optionally pretreated lignocellulosic material.

### Claim 5 (depends on 4)

The process according to claim 4, wherein the viscosity of the lignocellulosic material in the first container during (d) is kept below 1000 cP.

### Claim 6 (depends on 1)

The process according to claim 1, wherein an aqueous composition, optionally an aqueous enzyme composition comprising at least two cellulases, is added to the first container.

### Claim 7 (depends on 1)

The process according to claim 1, wherein the oxygen is added to a headspace of the container.

### Claim 8 (depends on 1)

The process according to claim 1, wherein the first container and/or second container has a volume of at least 1 m3.

### Claim 9 (depends on 1)

The process according to claim 1, wherein the enzymatic hydrolysis time in (d) is 3 to 24 hours.

### Claim 10 (depends on 1)

The process according to claim 1, wherein the enzymatic hydrolysis time in (f) is 3 to 120 hours.

### Claim 11 (depends on 1)

The process according to claim 1, wherein the enzymatic hydrolysis in (d) and/or (f) is conducted at a temperature of 45° C. or more, optionally 50° C. or more and optionally at a temperature of 55° C. or more.

### Claim 12 (depends on 1)

The process according to claim 1, wherein the enzyme composition is derived from a fungus, optionally a microorganism of the genus Rasamsonia, or the enzyme composition comprises a fungal enzyme, optionally a Rasamsonia enzyme.

### Claim 13 (depends on 2)

The process according to claim 2, wherein the fermentation in (h) is performed in the second container.

### Claim 14 (depends on 1)

The process according to claim 1, wherein the dry matter content at the end of hydrolysis of (f) is 5 wt % or higher.

### Claim 15 (depends on 1)

Process according to claim 1, wherein the enzyme composition is a whole fermentation broth.

### Claim 16 (depends on 2)

The process according to claim 2, wherein the fermentation in (h) is conducted with a microorganism that is able to ferment at least one C5 sugar.