# US 9,434,972 B2 · Processes for enzymatic refining of pretreated cellulosic material for saccharification > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,434,972 B2 - Document ID: US9434972B2 - Status: Granted - Kind: B2 - Application number: 13/976,785 - Filed: 2012-01-31 - Published: 2013-12-19 - Granted: 2016-09-06 - Earliest claimed: 2011-01-31 - Assignees: Novozymes North America, Inc. - Applicants: Armindo Ribiero Gaspar; Donald Higgins; Ye Chen; Hui Xu - Inventors: Armindo Ribiero Gaspar; Donald Higgins; Ye Chen; Hui Xu - Web page: https://patentgrep.com/patent/US9434972B2 ## CPC - C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} - C08H 8/00: Macromolecular compounds derived from lignocellulosic materials {(pretreatment thereof B27N)} - C12P 7/10: substrate containing cellulosic material - C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60) - C13K 1/02: obtained by saccharification of cellulosic materials (manufacture of fodder A23K10/32) - D21C 5/005: {Treatment of cellulose-containing material with microorganisms or enzymes} - 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 - C12Y 301/01073: Feruloyl esterase (3.1.1.73) - Y02E 50/10: Biofuels, e.g. bio-diesel - Y02P 20/52: using catalysts, e.g. selective catalysts - 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)} - C08H: DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS (polysaccharides C08B; natural rubber C08C; natural resins or their derivatives C09F; bituminous materials C10) - 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) - D21C: PRODUCTION OF CELLULOSE BY REMOVING NON-CELLULOSE SUBSTANCES FROM CELLULOSE-CONTAINING MATERIALS; REGENERATION OF PULPING LIQUORS; APPARATUS THEREFOR - C12Y: ENZYMES - Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION - Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS ## Abstract The present invention relates to processes for processes for enzymatic refining of a pretreated cellulosic material for saccharification. ## Claims ### Claim 1 A process for enzymatic refining of a pretreated cellulosic material, comprising: separating a liquor from the pretreated cellulosic material, contacting the liquor with feruloyl esterase to generate a treated liquor and recycling the treated liquor so that it is contacted with pretreated cellulosic material or refined pretreated cellulosic material. ### Claim 2 (depends on 1) The process of claim 1, where the feruloyl esterase comprises an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 136 or SEQ ID NO: 137, or fragments thereof with feruloyl esterase activity. ### Claim 3 (depends on 1) The process of claim 1, wherein the feruloyl esterase comprises or consists of SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 136 or SEQ ID NO: 137. ### Claim 4 (depends on 1) The process of claim 1, wherein the feruloyl esterase is one or more enzymes selected from the group consisting of FAE(1), FAE-A, FAE-C, and FAE-D. ### Claim 5 (depends on 1) The process of claim 1, further comprising recycling the liquor to a new pretreatment of a pretreated cellulosic material with the feruloyl esterase composition. ### Claim 6 (depends on 1) The process of claim 1 further comprising post-treating the refined pretreated cellulosic material with an enzymatic pre-treatment, chemical pre-treatment, mechanical pre-treatment and/or a physical pretreatment. ### Claim 7 (depends on 1) The process of claim 1, further comprising recovering the refined pretreated cellulosic material. ### Claim 8 (depends on 1) The process of claim 1, wherein the contacting with the esterase is performed with about 0.0005 to about 5 mg, about 0.001 to about 5 mg, about 0.0025 to about 5 mg, about 0.005 to about 5 mg, about 0.005 to about 4.5 mg, about 0.005 to about 4 mg, about 0.005 to about 3.5 mg, about 0.005 to about 3 mg, about 0.005 to about 2 mg, about 0.005 to about 1 mg, about 0.075 to about 1 mg, or about 0.1 to about 1 mg of esterase per g of pretreated cellulosic material. ### Claim 9 (depends on 1) The process of claim 1, wherein the contacting with esterase is performed with a total solids (TS) of about 1% to about 50%, about 2% to about 40%, about 2% to about 35%, about 3% to about 30%, about 3% to about 25%, about 4% to about 20%, or about 5% to about 10%. ### Claim 10 (depends on 1) The process of claim 1, wherein the contacting with esterase is performed at a pH of about 2 to about 9, about 3 to about 7.5, about 3.5 to about 7, about 4 to about 6.5, about 4.5 to about 6.5, about 4.5 to about 6.0, about 5 to about 6.0, or about 5 to about 5.5. ### Claim 11 (depends on 1) The process of claim 1, wherein the contacting is performed at a temperature in the range of about 20° C. to about 70° C., about 25° C. to about 65° C., about 30° C. to about 65° C., about 35° C. to about 65° C., about 40° C. to about 60° C., about 45° C. to about 55° C., or about 45° C. to about 50° C. ### Claim 12 (depends on 1) The process of claim 1, wherein the contacting with esterase is performed for a period of time of 5 minutes to 35 hours, 10 minutes to 15 hours, 10 hours to 15 hours, 10 hours to 20 hours, 20 hours to 24 hours, 24 hours to 30 hours, or 1 hour to 72 hours. ### Claim 13 A process for enzymatic refining of a pretreated cellulosic material, comprising: (a) processing the pretreated cellulosic material to form a solid/liquid mixture of pretreated cellulosic material; (b) separating a liquor from the solid/liquid mixture of pretreated cellulosic material; and (c) contacting the liquor with feruloyl esterase to generate a treated liquor.