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1. (WO2017147690) ENGINEERED BETA-GLUCOSIDASES AND METHODS OF USE THEREOF
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Pub. No.: WO/2017/147690 International Application No.: PCT/CA2017/050219
Publication Date: 08.09.2017 International Filing Date: 22.02.2017
IPC:
C12N 9/42 (2006.01) ,C12N 15/56 (2006.01) ,C12N 9/24 (2006.01) ,C12P 19/02 (2006.01) ,C12P 19/14 (2006.01) ,C12P 7/10 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9
Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14
Hydrolases (3.)
24
acting on glycosyl compounds (3.2)
42
acting on beta-1, 4-glucosidic bonds, e.g. cellulase
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
11
DNA or RNA fragments; Modified forms thereof
52
Genes encoding for enzymes or proenzymes
55
Hydrolases (3)
56
acting on glycosyl compounds (3.2), e.g. amylase, galactosidase, lysozyme
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9
Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14
Hydrolases (3.)
24
acting on glycosyl compounds (3.2)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19
Preparation of compounds containing saccharide radicals
02
Monosaccharides
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19
Preparation of compounds containing saccharide radicals
14
produced by the action of a carbohydrase, e.g. by alpha-amylase
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
7
Preparation of oxygen-containing organic compounds
02
containing a hydroxy group
04
acyclic
06
Ethanol, i.e. non-beverage
08
produced as by-product or from waste or cellulosic material substrate
10
substrate containing cellulosic material
Applicants:
VALORBEC SOCIÉTÉ EN COMMANDITE [CA/CA]; 355 Peel, Carrefour INGO Suite 503 Montréal, Québec H3C 2G9, CA
Inventors:
MARTIN, Vincent; CA
LARUE, Kane; CA
Agent:
GOUDREAU GAGE DUBUC; 2000, avenue McGill College #2200 Montréal, Québec H3A 3H3, CA
Priority Data:
62/302,37902.03.2016US
Title (EN) ENGINEERED BETA-GLUCOSIDASES AND METHODS OF USE THEREOF
(FR) BÊTA-GLUCOSIDASES MODIFIÉES ET PROCÉDÉS D'UTILISATION CORRESPONDANTS
Abstract:
(EN) A glycoside hydrolase family 3 (GH3) beta-glucosidase (BGL) polypeptide, comprising: (a) a GH3 BGL triosephosphateisomerase domain comprising a sequence as set forth in KGY1Y2Y3Y4LGP (SEQ ID NO: 251); (b) a GH3 BGL coordinating loop domain comprising a sequence as set forth in: GLDMX1MPX2X3X4X5X6X7X8X9X19X11X12X13X14X15X16 (SEQ ID NO: 252), except that at least one of X1to X16, when present, is smaller than a corresponding reference amino acid residue in a corresponding GH3 BGL reference coordinating loop domain comprising a sequence as set forth in GLDMX1refMPX2refX3refX4refX5refX6refX7refX8refX9refX19refX11refX12refX13refX14refX15refX16ref (SEQ ID NO: 252); (c) a GH3 BGL β/α sandwich domain comprising a sequence as set forth in: Z1Z2Z3Z4Z5Z6 (SEQ ID NO: 253); and/or (d) a GH3 BGL β/α sandwich domain comprising a sequence as set forth in: A1A2A3A4A5 (SEQ ID NO: 254) and a sequence as set forth in B1B2B3B4B5 (SEQ ID NO: 255), with the proviso that the GH3 BGL polypeptide is not a polypeptide as set forth in any one of sequences of SEQ ID NOs: 22 to 44, and is not a secreted form of the polypeptide of any one of SEQ ID NOs: 22 to 44. The GH3 BGL has an improved resistance to substrate inhibition and/or product inhibition. It may be used in methods of converting biomass into a fermentable sugar.
(FR) L'invention concerne un polypeptide de type bêta-glucosidase (BGL) de la famille des glycoside hydrolases 3 (GH3) comprenant a) : un domaine de triosephosphate-isomérase de BGL de GH3 comprenant une séquence telle qu'indiquée dans KGY1Y2Y3Y4LGP (SEQ ID NO : 251) ; b) un domaine de boucle de coordination de BGL de GH3 comprenant une séquence telle qu'indiquée dans : GLDMX1MPX2X3X4X5X6X7X8X9X19X11X12X13X14X15X16 (SEQ ID NO :252), sauf qu'au moins un des X1 à X16, lorsqu'ils sont présents, est plus petit qu'un résidu d'acide aminé de référence correspondant dans un domaine de boucle de coordination de BGL de GH3 de référence correspondant comprenant une séquence telle qu'indiquée dans GLDMX1refMPX2refX3refX4refX5refX6refX7refX8refX9refX19refX11refX12refX13refX14refX15refX16ref (SEQ ID NO : 252) ; (c) un domaine sandwich β/α de BGL de GH3 comprenant une séquence telle qu'indiquée dans : Z1Z2Z3Z4Z5Z6 (SEQ ID NO : 253) ; et/ou (d) un domaine sandwich β/α de BGL de GH3 comprenant une séquence telle qu'indiquée dans : A1A2A3A4A5 (SEQ ID NO : 254) et une séquence telle qu'indiquée dans B1B2B3B4B5 (SEQ ID NO :255), à condition que le polypeptide de BGL de GH3 ne soit pas un polypeptide tel qu'indiqué dans l'une quelconque des séquences SEQ ID NO : 22 à 44 et ne soit pas une forme sécrétée du polypeptide selon l'un quelconque des séquences SEQ ID NO : 22 à 44. La BGL de GH3 présente une résistance accrue à l'inhibition du substrat et/ou à l'inhibition du produit. Elle peut être utilisée dans des procédés de conversion de biomasse en un sucre fermentable.
Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: English (EN)
Filing Language: English (EN)