# US 10,865,426 B2 · Sequentially-fed process for enzymatic hydrolysis with additions of pre-treated substrate increasingly spread out over time > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,865,426 B2 - Document ID: US10865426B2 - Status: Granted - Kind: B2 - Application number: 16/143,562 - Filed: 2018-09-27 - Published: 2019-03-28 - Granted: 2020-12-15 - Earliest claimed: 2017-09-28 - Assignees: Ifp Energies Nouvelles - Applicants: IFP Energies Nouvelles - Inventors: Federico Battista; Romain Rousset; Melanie Gomez Almendros - Web page: https://patentgrep.com/patent/US10865426B2 ## CPC - C12P 7/10: substrate containing cellulosic material - 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) - 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)} - 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 present invention relates to a sequentially fed process for enzymatic hydrolysis in which, under agitation, a pre-treated lignocellulosic substrate is brought into contact with water and with enzymes in a reactor, said process being characterised in that the sequential addition to the reactor of the pre-treated lignocellulosic substrate is carried out increasingly spread out over time, in order to obtain a predetermined final content of dry matter. ## Claims ### Claim 1 A process for enzymatic hydrolysis of pre-treated lignocellulosic substrate, comprising bringing into contact, under agitation, at least one pre-treated lignocellulosic substrate, which comes from raw or treated wood, marine biomass or a by-product of agriculture, with water and with enzymes in a reactor, comprising adding the at least one pre-treated lignocellulosic substrate to the reactor in a sequential batch-wise manner such that intervals between additions of batches are longer over time within the same process, and obtaining a predetermined final content of dry matter that is higher than 12% by weight. ### Claim 2 (depends on 1) The process according to claim 1, in which the final content of dry matter is 17 to 30% by weight. ### Claim 3 (depends on 1) The process according to claim 1, in which the final content of dry matter is between 18 and 24% by weight. ### Claim 4 (depends on 1) The process according to claim 1, in which the addition of the enzymes to the reactor is carried out sequentially and increasingly spread out over time. ### Claim 5 (depends on 1) The process according to claim 1, in which the pre-treated lignocellulosic substrate and the enzymes are added at the same time to the reactor. ### Claim 6 (depends on 1) The process according to claim 1, in which, with each addition, the pre-treated lignocellulosic substrate is added in the same amount. ### Claim 7 (depends on 1) The process according to claim 1, in which, with each addition, the enzymes are added in the same amount. ### Claim 8 (depends on 1) The process according to claim 1, in which the reactor comprises a stirrer and the ratio of the diameter of the stirrer to the diameter of the reactor D:T is between 0.3 and 0.75. ### Claim 9 (depends on 1) The process according to claim 1, in which the reactor comprises a stirrer, which is an inclined blade impeller or marine impeller. ### Claim 10 (depends on 1) The process according to claim 1, in which the enzymes have a concentration of between 0.1 and 60 mg of enzymes per gram of cellulose. ### Claim 11 (depends on 1) The process according to claim 1, in which the process takes place at a temperature of between 40 and 60° C., at a pH of between 4 and 6, and at atmospheric pressure. ### Claim 12 (depends on 1) The process according to claim 1, in which various pre-treated lignocellulosic substrates are processed, on their own or as a mixture. ### Claim 13 (depends on 1) The process according to claim 1, which is followed by fermentation in the presence of an alcohol-producing microorganism. ### Claim 14 (depends on 1) The process according to claim 1, which is carried out in the presence of an alcohol-producing microorganism according to a process of simultaneous saccharification and fermentation. ### Claim 15 A process for enzymatic hydrolysis of pre-treated lignocellulosic substrate, comprising bringing into contact, under agitation, at least one pre-treated lignocellulosic substrate with water and with enzymes in a reactor, comprising adding the at least one pre-treated lignocellulosic substrate to the reactor in a sequential batch-wise manner such that intervals between additions of batches are longer over time within the same process, and obtaining a predetermined final content of dry matter that is higher than 12% by weight, wherein the at least one pre-treated lignocellulosic substrate comes from raw or treated wood, or a by-product of agriculture. ### Claim 16 A process for enzymatic hydrolysis of pre-treated lignocellulosic substrate, comprising bringing into contact, under agitation, at least one pre-treated lignocellulosic substrate, which is not from paper industry, with water and with enzymes in a reactor, comprising adding the at least one pre-treated lignocellulosic substrate to the reactor in a sequential batch-wise manner such that intervals between additions of batches are longer over time within the same process, and obtaining a predetermined final content of dry matter that is higher than 12% by weight. ### Claim 17 (depends on 16) The process according to claim 16, in which the final content of dry matter is 17 to 30% by weight.