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DK2367928T3 - Xyloseisomerasegener og deres anvendelse ved fermentering af pentosesukre - Google Patents

Xyloseisomerasegener og deres anvendelse ved fermentering af pentosesukre Download PDF

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DK2367928T3
DK2367928T3 DK09775365.1T DK09775365T DK2367928T3 DK 2367928 T3 DK2367928 T3 DK 2367928T3 DK 09775365 T DK09775365 T DK 09775365T DK 2367928 T3 DK2367928 T3 DK 2367928T3
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Aloysius Wilhelmus Rudolphus Hubertus Teunissen
Bont Johannes Adrianus Maria De
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Dsm Ip Assets Bv
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Claims (14)

1. Eukaryot mikrobiel celle, som omfatter nukleotidsekvenser, hvis ekspression giver cellen evnen eller øger evnen hos cellen til at isomerisere xylose direkte til xylulose, hvor nukleotidsekvensen koder for et polypeptid med xyloseisomeraseaktivitet, hvor polypeptidets aminosyresekvens: i) udviser mindst 70% sekvensidentitet med aminosyresekvensen ifølge mindst én af SEQ ID NO: 1, 2 og 35; ii) omfatter xyloseisomerase-signatursekvenserne VXW[GP]GREG[YSTA] i positionerne 187-195 og [LIVM]EPKPX[EQ]P i positionerne og 232-239, hvor "X" kan være en hvilken som helst aminosyre, og hvor aminosyrer i parentes angiver, at én af aminosyrerne i parentesen kan være til stede i den pågældende position i signatursekvensen; iii) omfatter de bevarede katalytiske aminosyrerester His-102, Asp-105 og Asp-340; og iv) omfatter et eller flere aminosyresekvenselementer valgt fra gruppen bestående af: (a) en methioninrest i position 91; (b) aminosyresekvensen TGIKLL i positionerne 134-139; (c) en phenylalaninrest i position 230; (d) aminosyrerne phenylalanin og lysin i henholdsvis position 264 og 2 65; (e) aminosyresekvensen TLAGH i positionerne 274-278; (f) aminosyresekvensen RYASF i positionerne 387-391; (g) en glycinrest i position 394; og (h) en alaninrest i position 431; hvor aminosyrepositionerne i ii) , iii) og i sekvenselementerne (a)-(h) henviser til positioner i reference-aminosyresekvensen ifølge SEQ ID NO: 3, og hvor polypeptidet ikke omfatter en aminosyresekvens, der udviser mere end 90% sekvensidentitet med en af SEQ ID NO: 3-7.
2. Celle ifølge krav 1, hvor nukleotidsekvensen koder for en aminosyresekvens, som er naturligt forekommende i en bakterie af slægterne Clostridium eller Fusobacterium eller et sækdyr af slægten Ciona.
3. Celle ifølge et hvilket som helst af de foregående krav, hvor cellen er en gær eller en filamentøs svamp af en slægt valgt fra gruppen bestående af Saccharomyces, Kluyveromyces, Candida, Pichia, Schizosaccharomyces, Hansenula, Kloeckera, Schwanniomyces, Yarrowia, Aspergillus, Trichoderma, Humicola, Acremonium, Fusarium og Penicillium.
4. Celle ifølge krav 3, hvor cellen er en gær, som er i stand til anaerob alkoholisk fermentering.
5. Celle ifølge krav 4, hvor gæren tilhører en art valgt fra gruppen bestående af S. cerevisiae, S. exiguus, S. bayanus, K. lactis, K. marxianus og Schizosaccharomyces pombe.
6. Celle ifølge et hvilket som helst af de foregående krav, hvor nukleotidsekvensen, som koder for polypeptidet med xyloseisomeraseaktivitet, er operativt forbundet til en promotor, der bevirker tilstrækkelig ekspression af nukleotidsekvenserne i cellen til at give cellen evnen til at isomerisere xylose til xylulose.
7. Celle ifølge et hvilket som helst af de foregående krav, hvor cellen endvidere omfatter en genetisk modifikation, som øger den specifikke xylulosekinaseaktivitet, hvor enzymet xylulosekinase overudtrykkes.
8. Celle ifølge et hvilket som helst af de foregående krav, hvor cellen omfatter en genetisk modifikation, som øger fluxen for pentosephosphatreaktionsvejen, hvor den genetiske modifikation omfatter overekspression af mindst ét enzym fra den ikke-oxidative del af pentosephosphatreaktionsvejen valgt fra gruppen bestående af enzymerne ribulose-5-phosphat-isomerase, ribulose-5-phosphat-3-epimerase, transketolase og transaldolase.
9. Celle ifølge et hvilket som helst af de foregående krav, hvor værtscellen omfatter en genetisk modifikation, som reducerer uspecifik aldosereduktaseaktivitet i cellen, hvor genet GRE3 er reduceret.
10. Celle ifølge et hvilket som helst af de foregående krav, hvor cellen udviser evnen til at omdanne L-arabinose til D-xylulose-5-phosphat, idet cellen omfatter nukleotidsekvenser, som koder for en arabinoseisomerase, en ribulokinase og en ribulose-5-P-4-epimerase.
11. Celle ifølge et hvilket som helst af de foregående krav, hvor cellen omfatter mindst én yderligere genetisk modifikation, som resulterer i en egenskab valgt fra gruppen bestående af: (a) øger transport af mindst én af xylose og arabinose ind i værtscellen; (b) nedsat følsomhed over for katabolitrepression; (c) øget tolerance over for ethanol, osmolaritet eller organiske syrer; og (d) reduceret fremstilling af biprodukter.
12. Celle ifølge et hvilket som helst af de foregående krav, hvor cellen har evnen til at fremstille mindst ét fermenteringsprodukt valgt fra gruppen bestående af ethanol, mælkesyre, 3-hydroxy-propionsyre, acrylsyre, eddikesyre, ravsyre, citronsyre, aminosyrer, 1,3-propandiol, ethylen, glycerol, smørsyre, caproat, butanol, glyoxylat, β-lactamantibiotika og cephalosporiner.
13. Fremgangsmåde til fremstilling af et fermenteringsprodukt valgt fra gruppen bestående af ethanol, mælkesyre, 3-hydroxy-propionsyre, acrylsyre, eddikesyre, ravsyre, citronsyre, aminosyrer, 1,3-propandiol, ethylen, glycerol, smørsyre, caproat, butanol, glyoxylat, β-lactamantibiotika og cephalosporiner, hvorved fremgangsmåden omfatter følgende trin: (a) fermentering af et medium, som indeholder en kilde til xylose og eventuelt en kilde til arabinose, med cellen som defineret i et hvilket som helst af kravene 1-12, hvorved cellen fermenterer xylose til fermenteringsproduktet, og eventuelt (b) indvinding af fermenteringsproduktet.
14. Fremgangsmåde ifølge krav 13, hvorved mediet også indeholder en kilde til glucose.
DK09775365.1T 2008-12-24 2009-12-24 Xyloseisomerasegener og deres anvendelse ved fermentering af pentosesukre DK2367928T3 (da)

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JP5843229B2 (ja) 2009-12-22 2016-01-13 株式会社豊田中央研究所 キシロースイソメラーゼ及びその利用
GB201010176D0 (en) 2010-06-17 2010-07-21 Bergen Teknologioverforing As Method
ES2499590T5 (es) 2010-07-07 2023-01-18 Novozymes North America Inc Proceso de fermentación con polipéptidos GH61
WO2012024046A2 (en) * 2010-08-20 2012-02-23 Codexis, Inc. Pentose fermentation by a recombinant microorganism
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ES2532508T3 (es) 2010-11-18 2015-03-27 Dsm Ip Assets B.V. Cepas de levaduras manipuladas para producir etanol a partir de glicerol
US9365857B2 (en) * 2010-12-23 2016-06-14 Archer Daniels Midland Company Xylose isomerase and xylitol dehydrogenase combination for xylose fermentation to ethanol and B. fragilis xylose isomerase
CN102174549B (zh) 2011-02-22 2012-10-10 山东大学 一种编码木糖异构酶的核酸分子及其编码的木糖异构酶
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