ES2542621B1 - Polypeptides with monooxygenase polysaccharide activity and their use for the production of fermentable sugars - Google Patents

Polypeptides with monooxygenase polysaccharide activity and their use for the production of fermentable sugars Download PDF

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Publication number
ES2542621B1
ES2542621B1 ES201430155A ES201430155A ES2542621B1 ES 2542621 B1 ES2542621 B1 ES 2542621B1 ES 201430155 A ES201430155 A ES 201430155A ES 201430155 A ES201430155 A ES 201430155A ES 2542621 B1 ES2542621 B1 ES 2542621B1
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Prior art keywords
host cell
bioproduct
cellulosic biomass
polypeptides
fermentable sugars
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Expired - Fee Related
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ES201430155A
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Spanish (es)
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ES2542621A1 (en
Inventor
Bruno DÍEZ GARCÍA
Ana GÓMEZ RODRÍGUEZ
Jorge GIL MARTÍNEZ
Noelia VALBUENA CRESPO
Francisco Manuel REYES SOSA
Antonio Javier MORENO PÉREZ
Rafael DUEÑAS SÁNCHEZ
Ana María MUÑOZ GONZÁLEZ
Dolores PÉREZ GÓMEZ
Sandra GAVALDÁ MARTÍN
Lucía MARTÍN PÉREZ
Laura SÁNCHEZ ZAMORANO
Consolación ÁLVAREZ NÚÑEZ
María De Los Ángeles BERMÚDEZ ALCÁNTARA
Laura LEDESMA GARCÍA
Ana Isabel PLATERO GÓMEZ
Pablo GUTIÉRREZ GÓMEZ
Ricardo Arjona Antolín
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Abengoa Bioenergia Nuevas Technologias SA
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Abengoa Bioenergia Nuevas Technologias SA
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Priority to ES201430155A priority Critical patent/ES2542621B1/en
Priority to PCT/ES2015/070079 priority patent/WO2015118205A1/en
Publication of ES2542621A1 publication Critical patent/ES2542621A1/en
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Publication of ES2542621B1 publication Critical patent/ES2542621B1/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C12N9/0083—Miscellaneous (1.14.99)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/06—Ethanol, i.e. non-beverage
    • C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
    • C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P2203/00—Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Polipéptidos con actividad polisacárido monooxigenasa y su uso para la producción de azúcares fermentables.#La invención se refiere a polipéptidos con actividad polisacárido monooxigenasa, a una célula hospedadora que los expresa, preferiblemente una célula de Myceliophthora thermophila, a una composición enzimática que comprende al menos uno de dichos polipéptidos, preferiblemente junto con otras enzimas celulolíticas, al uso de esta célula hospedadora, de al menos uno de los polipéptidos con actividad polisacárido monooxigenasa o de la composición enzimática para la degradación de biomasa celulósica y a un procedimiento para producir bioproductos, preferiblemente bioetanol, que comprende el uso de dicha célula hospedadora, de al menos uno de los polipéptidos de la invención o de la composición enzimática de la invención.Polypeptides with monooxygenase polysaccharide activity and their use for the production of fermentable sugars. # The invention relates to polypeptides with monooxygenase polysaccharide activity, to a host cell that expresses them, preferably a Myceliophthora thermophila cell, to an enzyme composition comprising at least one of said polypeptides, preferably together with other cellulolytic enzymes, to the use of this host cell, of at least one of the polypeptides with polysaccharide monooxygenase activity or of the enzymatic composition for the degradation of cellulosic biomass and to a method for producing bioproducts, preferably bioethanol , which comprises the use of said host cell, of at least one of the polypeptides of the invention or of the enzymatic composition of the invention.

Description

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Claims (13)

ES 2542621 Bl ES 2542621 Bl REIVINDICACIONES
1. one.
Construcción génica que comprende un polinucleótido que codifica el polipéptido que consiste en la SEO ID NO: 43, y otro polinucleótido que codifica el polipéptido que consiste en la SEO ID NO: 44. Gene construct comprising a polynucleotide encoding the polypeptide consisting of SEO ID NO: 43, and another polynucleotide encoding the polypeptide consisting of SEO ID NO: 44.
2. 2.
Construcción génica según la reivindicación 1, donde dicha construcción génica es un vector de expresión. Gene construct according to claim 1, wherein said gene construct is an expression vector.
3. 3.
Célula hospedadora que comprende la construcción génica según cualquiera de las reivindicaciones 1 ó 2. Host cell comprising the gene construct according to either of claims 1 or 2.
4. Four.
Célula hospedadora según la reivindicación 3, donde la célula hospedadora es Myceliophthora thermophila C1. Host cell according to claim 3, wherein the host cell is Myceliophthora thermophila C1.
5. 5.
Composición enzimática que comprende un polipéptido que consiste en la secuencia aminoacídica SEO ID NO: 43 y otro polipéptido que consiste en la secuencia aminoacídica SEO ID NO: 44 y, preferiblemente, otra enzima celulolítica seleccionada de la lista que consiste en: endoglucanasa, beta-glucosidasa, celobiohidrolasa, beta-xilosidasa, endoxilanasa, endoxiloglucanasa, polisacárido monooxigenasa o cualquier combinación de las mismas. Enzyme composition comprising a polypeptide consisting of the amino acid sequence SEO ID NO: 43 and another polypeptide consisting of the amino acid sequence SEO ID NO: 44 and preferably another cellulolytic enzyme selected from the list consisting of: endoglucanase, beta- glucosidase, cellobiohydrolase, beta-xylosidase, endoxylanase, endoxyloglucanase, polysaccharide monooxygenase, or any combination thereof.
6. 6.
Composición enzimática según la reivindicación 5, donde dicha composición es una mezcla enzimática expresada por la célula según cualquiera de las reivindicaciones 3 ó 4. Enzyme composition according to claim 5, wherein said composition is an enzyme mixture expressed by the cell according to any of claims 3 or 4.
7. 7.
Uso de la célula hospedadora según cualquiera de las reivindicaciones 3 ó 4, o de la composición según cualquiera de las reivindicaciones 5 ó 6, para la degradación de biomasa celulósica. Use of the host cell according to any of claims 3 or 4, or of the composition according to any of claims 5 or 6, for the degradation of cellulosic biomass.
8. 8.
Uso según la reivindicación 7, donde la degradación de biomasa celulósica tiene lugar en un proceso de producción de un bioproducto. Use according to claim 7, where the degradation of cellulosic biomass takes place in a production process of a bioproduct.
9. 9.
Uso según la reivindicación 8, donde el bioproducto es un biocombustible. Use according to claim 8, where the bioproduct is a biofuel.
10. 10.
Uso según la reivindicación 9, donde el biocombustible es bioetanol. Use according to claim 9, where the biofuel is bioethanol.
ES 2542621 Bl ES 2542621 Bl 11. Procedimiento para producir azúcares fermentables que comprende los siguientes pasos: a) incubar biomasa celulósica con la composición según cualquiera de las 5 reivindicaciones 5 ó 6, y 11. Process for producing fermentable sugars comprising the following steps: a) incubating cellulosic biomass with the composition according to any of claims 5 or 6, and b) recuperar los azúcares fermentables obtenidos después de la incubación del paso (a). b) recover the fermentable sugars obtained after incubation in step (a). 12. Procedimiento para producir un bioproducto a partir de biomasa celulósica que 10 comprende los siguientes pasos: a) incubar biomasa celulósica con la composición según cualquiera de las reivindicaciones 5 ó 6, b) fermentar los azúcares fermentables obtenidos después de la incubación 12. Procedure for producing a bioproduct from cellulosic biomass comprising the following steps: a) incubating cellulosic biomass with the composition according to any of claims 5 or 6, b) fermenting the fermentable sugars obtained after incubation del paso (a) con al menos un microorganismo fermentador, y 15 c) recuperar el bioproducto obtenido después de la fermentación del paso (b). of step (a) with at least one fermenting microorganism, and 15 c) recovering the bioproduct obtained after the fermentation of step (b). 13. El procedimiento de acuerdo con la reivindicación 12, donde el bioproducto es un biocombustible. 13. The method according to claim 12, wherein the bioproduct is a biofuel. 20 14. El procedimiento de acuerdo con la reivindicación 13, donde el biocombustible es bioetanol. The method according to claim 13, where the biofuel is bioethanol. imagen1image 1
ES201430155A 2014-02-07 2014-02-07 Polypeptides with monooxygenase polysaccharide activity and their use for the production of fermentable sugars Expired - Fee Related ES2542621B1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
ES201430155A ES2542621B1 (en) 2014-02-07 2014-02-07 Polypeptides with monooxygenase polysaccharide activity and their use for the production of fermentable sugars
PCT/ES2015/070079 WO2015118205A1 (en) 2014-02-07 2015-02-05 Polypeptides with polysaccharide monooxygenase activity and use thereof for the production of fermentable sugars

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ES201430155A ES2542621B1 (en) 2014-02-07 2014-02-07 Polypeptides with monooxygenase polysaccharide activity and their use for the production of fermentable sugars

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ES2542621B1 true ES2542621B1 (en) 2016-08-16

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3315609A1 (en) 2015-06-23 2018-05-02 Abengoa Bioenergía Nuevas Tecnologías, S. A. Cellulolytic compositions comprising monooxygenase polysaccharide enzymes with improved activity
CN106755011A (en) * 2015-11-25 2017-05-31 中国科学院大连化学物理研究所 A kind of polysaccharide cracks monooxygenase LPMO M2 encoding genes and its enzyme and preparation method and application

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* Cited by examiner, † Cited by third party
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DE69922978T2 (en) 1998-10-06 2005-12-08 Emalfarb, Mark Aaron, Jupiter TRANSFORMATION SYSTEM IN FILAMENTOUS FUNGICIDES CHRYSOSPORIUM HOST CELLS
KR20020026456A (en) 2000-04-13 2002-04-10 에말파브 마크 아론 High-throughput screening of expressed dna libraries in filamentous fungi
DK2606131T3 (en) * 2010-08-20 2017-06-19 Codexis Inc Use of proteins from glycoside hydrolase 61 family for cellulose processing
BR112014004171A2 (en) * 2011-08-22 2018-11-06 Codexis Inc gh61 glycoside hydrolase protein variants and cofactors that increase gh61 activity
BR112014006676A2 (en) * 2011-09-20 2018-09-04 Codexis Inc endoglucanase 1b
EP2760997A4 (en) * 2011-09-30 2015-02-11 Codexis Inc Fungal proteases
CA2879682A1 (en) * 2012-07-19 2014-01-30 Novozymes A/S Methods for increasing enzymatic hydrolysis of cellulosic material

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