# US 10,479,984 B2 · Polypeptides having cellobiohydrolase activity and polynucleotides encoding same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,479,984 B2 - Document ID: US10479984B2 - Status: Granted - Kind: B2 - Application number: 15/926,792 - Filed: 2016-09-22 - Published: 2018-08-09 - Granted: 2019-11-19 - Earliest claimed: 2015-09-22 - Assignees: Novozymes A/S - Applicants: Novozymes A/S - Inventors: Lan Tang; Xianzhi Jiang; Bucong Han; Christine Kai; Ye Liu - Web page: https://patentgrep.com/patent/US10479984B2 ## CPC - C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)} - C12N 15/8257: {for the production of primary gene products, e.g. pharmaceutical products, interferon} - C12Y 302/01091: Cellulose 1,4-beta-cellobiosidase (3.2.1.91) - C12Y 302/01176: Cellulose 1,4-beta-cellobiosidase (reducing end)(3.2.1.176) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Y: ENZYMES ## Abstract Disclosed herein are polypeptides having cellobiohydrolase activity, catalytic domains, carbohydrate binding modules and polynucleotides encoding the polypeptides, catalytic domains or carbohydrate binding modules. Disclosed herein are nucleic acid constructs, vectors and host cells comprise the polynucleotides as well as their uses, and methods of producing the polypeptides, catalytic domains, carbohydrate binding modules. ## Claims ### Claim 1 A polypeptide comprising a carbohydrate binding module operably linked to a catalytic domain, wherein the carbohydrate binding module is heterologous to the polypeptide, and wherein the carbohydrate binding module is selected from the group consisting of: (a) a carbohydrate binding module having at least 95% sequence identity to amino acids 517 to 545 of SEQ ID NO: 2; (b) a carbohydrate binding module comprising amino acids 517 to 545 of SEQ ID NO: 2; and (c) a carbohydrate binding module encoded by a polynucleotide having at least 98% sequence identity to nucleotides 1549 to 1635 of SEQ ID NO: 1. ### Claim 2 A nucleic acid construct or expression vector comprising a polynucleotide encoding a polypeptide having cellobiohydrolase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct the production of the polypeptide in a recombinant host cell, and wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of: (a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2; (b) a polypeptide encoded by a polynucleotide having at least 98% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1; (c) a polypeptide comprising a catalytic domain having at least 95% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2; and (d) a polypeptide comprising a catalytic domain encoded by a polynucleotide having at least 98% sequence identity to nucleotides 76 to 1392 of SEQ ID NO: 1. ### Claim 3 (depends on 2) A recombinant host cell transformed with the nucleic acid construct or expression vector of claim 2. ### Claim 4 (depends on 3) A method of producing a polypeptide having cellobiohydrolase activity, comprising cultivating the recombinant host cell of claim 3 under conditions conducive for production of the polypeptide, and optionally recovering the polypeptide. ### Claim 5 (depends on 2) A transgenic plant, plant part or plant cell transformed with the nucleic acid construct or expression vector of claim 2. ### Claim 6 (depends on 5) A method of producing a polypeptide having cellobiohydrolase activity, comprising cultivating the transgenic plant or plant cell of claim 5 under conditions conducive for production of the polypeptide, and optionally recovering the polypeptide. ### Claim 7 A process for degrading a cellulosic or hemicellulosic material, comprising: treating the cellulosic or hemicellulosic material with an enzyme composition comprising a polypeptide having cellobiohydrolase activity, wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of: (a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2; (b) a polypeptide encoded by a polynucleotide having at least 98% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1; (c) a polypeptide comprising a catalytic domain having at least 95% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2; and (d) a polypeptide comprising a catalytic domain encoded by a polynucleotide having at least 98% sequence identity to nucleotides 76 to 1392 of SEQ ID NO: 1. ### Claim 8 (depends on 7) The process of claim 7, wherein the enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, an AA9 polypeptide, a hemicellulase, a cellulose inducible protein (CIP) an esterase, an expansin, a ligninolytic enzyme, an oxidoreductase, a pectinase, a protease, and a swollenin. ### Claim 9 (depends on 7) The process of claim 7, further comprising recovering the degraded cellulosic or hemicellulosic material. ### Claim 10 A process for producing a fermentation product, comprising: (a) saccharifying a cellulosic or hemicellulosic material with an enzyme composition comprising a polypeptide having cellobiohydrolase activity, wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of: (i) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2; (ii) a polypeptide encoded by a polynucleotide having at least 98% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1; (iii) a polypeptide comprising a catalytic domain having at least 95% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2; and (iv) a polypeptide comprising a catalytic domain encoded by a polynucleotide having at least 98% sequence identity to nucleotides 76 to 1392 of SEQ ID NO: 1; (b) fermenting the saccharified cellulosic or hemicellulosic material with one or more fermenting microorganisms to produce the fermentation product; and (c) recovering the fermentation product from the fermentation. ### Claim 11 (depends on 10) The process of claim 10, wherein the enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, an AA9 polypeptide, a hemicellulase, a CIP, an esterase, an expansin, a ligninolytic enzyme, an oxidoreductase, a pectinase, a protease, and a swollenin. ### Claim 12 A process of fermenting a cellulosic or hemicellulosic material, comprising: fermenting the cellulosic or hemicellulosic material with one or more fermenting microorganisms, wherein the cellulosic or hemicellulosic material is saccharified with an enzyme composition comprising a polypeptide having cellobiohydrolase activity, wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of: (a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2; (b) a polypeptide encoded by a polynucleotide having at least 98% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1; (c) a polypeptide comprising a catalytic domain having at least 95% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2; and (d) a polypeptide comprising a catalytic domain encoded by a polynucleotide having at least 98% sequence identity to nucleotides 76 to 1392 of SEQ ID NO: 1. ### Claim 13 (depends on 12) The process of claim 12, wherein the enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, an AA9 polypeptide, a hemicellulase, a CIP, an esterase, an expansin, a ligninolytic enzyme, an oxidoreductase, a pectinase, a protease, and a swollenin. ### Claim 14 (depends on 12) The process of claim 12, wherein the fermenting of the cellulosic or hemicellulosic material produces a fermentation product. ### Claim 15 (depends on 14) The process of claim 14, further comprising recovering the fermentation product from the fermentation. ### Claim 16 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity comprises SEQ ID NO: 2 or the mature polypeptide of SEQ ID NO: 2. ### Claim 17 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity consists of SEQ ID NO: 2 or the mature polypeptide of SEQ ID NO: 2. ### Claim 18 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the mature polypeptide is amino acids 26 to 549 of SEQ ID NO: 2. ### Claim 19 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity comprises the catalytic domain of amino acids 26 to 464 of SEQ ID NO: 2. ### Claim 20 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity comprising a catalytic domain having at least 95% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2 further comprises a carbohydrate binding module. ### Claim 21 A recombinant host cell transformed with a nucleic acid construct or an expression vector comprising a polynucleotide encoding a polypeptide having cellobiohydrolase activity, wherein the polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide in the recombinant host cell, wherein the polypeptide having cellobiohydrolase activity is heterologous to the recombinant host cell, and wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of: (a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2; (b) a polypeptide encoded by a polynucleotide having at least 98% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1; (c) a polypeptide comprising a catalytic domain having at least 95% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2; and (d) a polypeptide comprising a catalytic domain encoded by a polynucleotide having at least 98% sequence identity to nucleotides 76 to 1392 of SEQ ID NO: 1. ### Claim 22 (depends on 21) A method of producing a polypeptide having cellobiohydrolase activity, comprising cultivating the recombinant host cell of claim 21 under conditions conducive for production of the polypeptide, and optionally recovering the polypeptide. ### Claim 23 (depends on 1) The polypeptide of claim 1, wherein the carbohydrate binding module has at least 96% sequence identity to amino acids 517 to 545 of SEQ ID NO: 2. ### Claim 24 (depends on 1) The polypeptide of claim 1, wherein the carbohydrate binding module has at least 97% sequence identity to amino acids 517 to 545 of SEQ ID NO: 2. ### Claim 25 (depends on 1) The polypeptide of claim 1, wherein the carbohydrate binding module has at least 98% sequence identity to amino acids 517 to 545 of SEQ ID NO: 2. ### Claim 26 (depends on 1) The polypeptide of claim 1, wherein the carbohydrate binding module has at least 99% sequence identity to amino acids 517 to 545 of SEQ ID NO: 2. ### Claim 27 (depends on 1) The polypeptide of claim 1, wherein the carbohydrate binding module comprises amino acids 517 to 545 of SEQ ID NO: 2. ### Claim 28 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2. ### Claim 29 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2. ### Claim 30 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2. ### Claim 31 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2. ### Claim 32 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity comprises a catalytic domain having at least 96% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2. ### Claim 33 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity comprises a catalytic domain having at least 97% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2. ### Claim 34 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity comprises a catalytic domain having at least 98% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2. ### Claim 35 (depends on 2) The nucleic acid construct or expression vector of claim 2, wherein the polypeptide having cellobiohydrolase activity comprises a catalytic domain having at least 99% sequence identity to amino acids 26 to 464 of SEQ ID NO: 2. ### Claim 36 (depends on 7) The process of claim 7, wherein the polypeptide having cellobiohydrolase activity comprises SEQ ID NO: 2 or the mature polypeptide of SEQ ID NO: 2. ### Claim 37 (depends on 7) The process of claim 7, wherein the polypeptide having cellobiohydrolase activity comprises the catalytic domain of amino acids 26 to 464 of SEQ ID NO: 2. ### Claim 38 (depends on 7) The process of claim 7, wherein the cellulosic or hemicellulosic material is pretreated. ### Claim 39 (depends on 10) The process of claim 10, wherein the polypeptide having cellobiohydrolase activity comprises SEQ ID NO: 2 or the mature polypeptide of SEQ ID NO: 2. ### Claim 40 (depends on 10) The process of claim 10, wherein the polypeptide having cellobiohydrolase activity comprises the catalytic domain of amino acids 26 to 464 of SEQ ID NO: 2. ### Claim 41 (depends on 10) The process of claim 10, wherein the cellulosic or hemicellulosic material is pretreated. ### Claim 42 (depends on 10) The process of claim 10, wherein steps (a) an (b) are performed simultaneously in a simultaneous saccharification and fermentation. ### Claim 43 (depends on 12) The process of claim 12, wherein the polypeptide having cellobiohydrolase activity comprises SEQ ID NO: 2 or the mature polypeptide of SEQ ID NO: 2. ### Claim 44 (depends on 12) The process of claim 12, wherein the polypeptide having cellobiohydrolase activity comprises the catalytic domain of amino acids 26 to 464 of SEQ ID NO: 2. ### Claim 45 (depends on 12) The process of claim 12, wherein the cellulosic or hemicellulosic material is pretreated.