DK2069490T4 - Amylaser og glucoamylaser, nukleinsyrer, der koder for dem, og fremgangsmåder til fremstilling og anvendelse deraf - Google Patents
Amylaser og glucoamylaser, nukleinsyrer, der koder for dem, og fremgangsmåder til fremstilling og anvendelse deraf Download PDFInfo
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- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
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- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
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- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2428—Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
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Claims (23)
1. Isoleret, syntetisk eller rekombinant nukleinsyre, som koder for et polypep-tid med glucoamylase-aktivitet, omfattende (a) en nukleinsyresekvens, der er mindst 90 % identisk med SEQ ID NO: 25; (b) et fragment af nukleinsyren af SEQ ID No. 25, hvor fragmentet koder for et polypeptid med glucoamylase-aktivitet; (c) nukleinsyren af (a) eller (b), som koder for et polypeptid med en glucoamylase-aktivitet, og yderligere omfattende en eller flere heterologe sekvenser, (d) nukleinsyresekvensen af (c), hvor den ene eller flere heterologe sekvenser omfatter en sekvens, som koder for en heterolog signalsekvens, et kulhydratbindende modul, katalytisk domæne (CD), eller en kombination deraf, hvor den heterologe signalsekvens, det kulhydratbindende modul eller den katalytiske domæne (CD) eventuelt er afledt fra et amylase-enzym eller et ikke-amylase-enzym; eller (e) en nukleinsyresekvens, der er fuldstændig komplementær til (a), (b), (c) eller (d).
2. Isoleret, syntetisk eller rekombinant nukleinsyre ifølge krav 1, som koder for et polypeptid med glucoamylase-aktivitet, hvor glucoamylase-aktiviteten omfatter hydrolysering af glucosidiske bindinger, eller eventuelt hydrolysering af glucosidiske bindinger i en polysaccharid, en oligosaccharid eller en stivelse, eller eventuelt omfatter de glucosidiske bindinger en a-1,4-glucosidisk binding eller en a-1,6-glucosidisk binding, eller hvor glucoamylase-aktiviteten omfatter spaltning af en maltose- eller en D-glucose-enhed fra en ikke-reducerende ende af en polysaccharid, en oligosaccharid eller en stivelse.
3. Vektor, ekspressionskassette eller kloningsvehikel omfattende: (a) en nukleinsyre, der omfatter nukleinsyresekvensen ifølge krav 1; (b) vektoren, ekspressionskassetten eller kloningsvehiklet af (a), hvor nukleinsyren er operabelt forbundet med en promotor; (c) vektoren, ekspressionskassetten eller kloningsvehiklet af (b), hvor promotoren er en plantepromotor eller en viral promotor; (d) vektoren, ekspressionskassetten eller kloningsvehiklet af (c), hvor promotoren er en inducerbar promotor eller en vævsspecifik promotor; eller (e) vektoren, ekspressionskassetten eller kloningsvehiklet af (d), hvor den vævsspecifikke promotor eventuelt er frø-specifik, en endosperm-specifik promoter, en blad-specifik, en rod-specifik, en stam-specifik eller en afskærings-induceret promoter.
4. Transformeret celle, omfattende en nukleinsyre omfattende nukleinsyrese-kvensen ifølge krav 1, vektor, ekspressionskassette eller kloningsvehikel ifølge krav 3, hvor cellen er en bakteriecelle, en pattedyrscelle, en gærcelle, en insektcelle eller en plantecelle, hvor plantecellen eventuelt er en kartoffel-, ris-, majs-, hvede-, tobak- eller bygcelle.
5. Transgen plante eller plantecelle, frø, planteprodukt, plantevæv eller plantedel, omfattende nukleinsyresekvens ifølge krav 1, eller vektor, ekspressionskassette eller kloningsvehikel ifølge krav 3; hvor planten, eller plantecellen, frøet, plante-produktet, plantevævet eller plantedelen er eller er afledt fra Anacardium, Andropogoneae, Arachis, Asparagus, Atropa, Avena, Beta, Brassica, Ceiba, Citrus, Citrullus, Capsicum, Carthamus, Chenopodiaceae, Cocos, Coffea, Cucumis, Cucurbita, Daucus, Elaeis, Fragaria, Glycine, Gossypium, Grainier, Helianthus, Heterocallis, Hordeum, Hyoscyamus, Lactuca, Linum, Lolium, Lupinus, Lycopersicon, Malus, Manihot, Majorana, Medicago, Nicotiana, Olea, Oryza, Panieum, Pannisetum, Persea, Phaseolus, Pistachia, Pisum, Ponceau, Pyrus, Prunus, Raphanus, Ricinus, Secale, Senecio, Sinapis, Solanum, Sorghum, Theobromus, Trigonella, Triticum, Vicia, Vitis, Vigna eller Zea, hvor planten eventuelt er (a) en tokimbladet plante (en dicot) eller en enkimbladet plante (en monocot); (b) majs, sojabønne, sukkerrør, sukkerroe, præriehirse, Miscanthus, hvede, Brassica, kassava, bomuld og byg; (c) en hirseplante, palme, solsikke, ris, byg, en peanut-plante, en sesam, en kartoffelplante, en tomatplante, en olieholdig plante, en rapsplante, en sojabønneplante, en græs- eller en tobaksplante; eller (d) arter fra familien eller slægterne Anacardium, Andropogoneae, Arachis, Asparagus, Atropa, Avena, Beta, Brassica, Ceiba, Citrus, Citrullus, Capsicum, Carthamus, Chenopodiaceae, Cocos, Coffea, Cucumis, Cucurbita, Daucus, Elaeis, Fragaria, Glycine, Gossypium, Helianthus, Heterocallis, Hordeum, Hyoscyamus, Lactuca, Linum, Lolium, Lupinus, Lycopersicon, Malus, Manihot, Majorana, Medicago, Nicotiana, Olea, Oryza, Panieum, Panni-setum, Persea, Phaseolus, Pistachia, Pisum, Ponceau, Pyrus, Prunus, Raphanus, Ricinus, Secale, Senecio, Sinapis, Solanum, Sorghum, Theobro-mus, Trigonella, Triticum, Vicia, Vitis, Vigna og/eller Zea, hvor det transgene frø eventuelt er et majsfrø, et hvedekorn, et olieholdigt frø, et rapsfrø, et sojabønnefrø, en palmekerne, et solsikkefrø, et sesamfrø, et ris-, et byg-, et peanut- eller et tobaksplantefrø, omfattende nukleinsyresekvensen ifølge krav 1, eller vektor, ekspressionskassette eller kloningsvehikel ifølge krav 3.
6. Isoleret, syntetisk eller rekombinant polypeptid, som har glucoamylase-aktivitet, omfattende (i) en aminosyresekvens, der har mindst 95 % sekvensidentitet med sekvensen med SEQ ID NO: 26; (ii) en aminosyresekvens, der er kodet af nukleinsyren ifølge krav 1; (iii) polypeptidet af (i) eller (ii), yderligere omfattende en eller flere heterologe sekvenser; (iv) polypeptidet af (iii) hvor den ene eller flere heterologe sekvenser omfatter en heterolog signalsekvens, et kulhydratbindende modul, et katalytisk domæne (CD), eller en kombination deraf, eller den heterologe sekvens omfatter en heterolog epitop, rensningstag eller etikette; (v) polypeptidet af (iv), hvor polypeptidet omfatter mindst ét glykolsylerings-sted, hvor glykosyleringen eventuelt er en N-forbundet glykosylering, hvor polypeptidet eventuelt glykosyleres efter at være eksprimeret i en P. pastoris eller en S. pombe; eller (vi) polypeptidet af (i), (ii), (iii), (iv) eller (v), hvor polypeptidet kan anvendes til at generere antistoffer, som binder specifikt til polypeptidet SEQ ID NO:26.
7. Isoleret, syntetisk eller rekombinant nukleinsyre ifølge krav 6, hvor polypeptidet har en glucoamylase-aktivitet, og hvor glucoamylase-aktiviteten omfatter hydrolysering af glucosidiske bindinger, eller eventuelt omfatter de glucosidiske bindinger en a-1,4-glucosidisk binding eller en a-1,6-glucosidisk binding, og/eller hvor glucoamylase-aktiviteten omfatter hydrolysering af glucosidiske bindinger i en polysaccharid, en oligosaccharid eller en stivelse, og/eller glucoamylase-aktiviteten omfatter spaltning af en maltose- eller en D-glucose-enhed fra en ikke-reducerende ende af en polysaccharid, en oligosaccharid eller en stivelse.
8. Fremgangsmåde til hydrolyse, fjerne eller gøre en polysaccharid, olig-osaccharid eller stivelse flydende, omfattende de følgende trin: (a) tilvejebringe polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe en sammensætning omfattende polysaccharid, oligosaccha-rid eller stivelse; og (c) bringe polypeptidet af trin (a) i kontakt med sammensætningen af trin (b) under forhold, hvor polypeptidet hydrolyserer, fjerne eller gør polysacchari-den, oligosacchariden eller stivelsen flydende.
9. Fremgangsmåde til at vaske en genstand, omfattende følgende trin: (a) tilvejebringe en sammensætning omfattende polypeptidet af krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe en genstand; og (c) bringe sammensætningen af trin (a) i kontakt med genstanden af trin (b) under forhold, hvor sammensætningen kan vaske genstanden.
10. Fremgangsmåde til hydrolyse af en polysaccharid, oligosaccharid eller stivelse i et foder eller en fødevare inden fortæring af et dyr, omfattende de følgende trin: (a) opnå et fodermateriale omfattende en stivelse, hvor polysaccharid, oligosaccharid eller stivelse i polysacchariden, oligosacchariden eller stivelsen kan hydrolyseres af polypeptidet ifølge krav 6, eller et polypeptid, som kodes af nukleinsyren ifølge krav 1; og (b) tilsætte polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1, til foder- eller fødevarematerialet i tilstrækkelige mængder og i tilstrækkelig tid til at bevirke hydrolyse af polysacchariden, oligosacchariden eller stivelsen og dannelse af et behandlet foder eller en behandlet fødevare, og derved hydrolysere polysacchariden, oligosacchariden eller stivelsen foderet eller fødevaren inden fortæring af dyret, hvor foderet eller fødevaren eventuelt omfatter ris, majs, byg, hvede, grøntsager eller kartoffel.
11. Fremgangsmåde til krympning af tekstiler, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe et tekstil; og (c) bringe polypeptidet af trin (a) i kontakt med tekstilet af trin (b) under forhold, hvor glucoamylasen kan krympe tekstilet.
12. Fremgangsmåde til behandling af lignocellulosiske fibre, omfattende de følgende trin: (a) tilvejebringe polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe en lignocellulosisk fiber; og (c) bringe polypeptidet af trin (a) i kontakt med fiberen af trin (b) under forhold, hvor polypeptidet kan behandle fiberen og derved forbedre fiberegenskaber.
13. Fremgangsmåde til at fremstille en sirup med højt maltoseindhold eller glucoseindhold, omfattende de følgende trin: (a) tilvejebringe polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe en sammensætning omfattende en polysaccharid, oligosac-charid eller stivelse; hvor polysacchariden, oligosacchariden eller stivelsen eventuelt er fra ris, majs, byg, hvede, grøntsager, kartoffel eller søde kartofler, og (c) bringe polypeptidet af trin (a) i kontakt med tekstilet af trin (b) under forhold, hvor polypeptidet af trin (a) kan hydrolysere sammensætningen af trin (b), og derved fremstille en sirup med højt maltoseindhold eller glucoseindhold.
14. Fremgangsmåde til forbedring af strømmen af polysaccharid-, oligosac-charid- eller stivelsesholdige produktionsfluider, omfattende de følgende trin: (a) tilvejebringe polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe produktionsfluid omfattende en polysaccharid, oligosaccharid eller stivelse; hvor produktionsfluiden eventuelt er fra en underjordisk formation; og (c) bringe polypeptidet af trin (a) i kontakt med produktionsfluiden af trin (b) under forhold, hvor glucoamylasen kan hydrolysere polysacchariden, olig- osacchariden eller stivelsen og derved forbedre strømmen deraf ved at forøge densiteten.
15. Fremgangsmåde til ændring af viskositeten af en sammensætning, omfattende: (') (a) tilvejebringe sammensætningen og polypeptidet af krav 6, eller et po-lypeptid, der kodes af nukleinsyren ifølge krav 1; og (a) behandle sammensætningen med polypeptidet af krav 6, eller det po-lypeptid, der kodes af nukleinsyren ifølge krav 1; eller (ii) fremgangsmåden af (i), hvor sammensætningen omfatter jord eller boreslam.
16. Fremgangsmåde til anvendelse af en glucoamylase i brygning eller fremstilling af alkohol, omfattende de følgende trin: (a) tilvejebringe polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe en sammensætning anvendt til brygning eller fremstilling af alkohol, omfattende en polysaccharid, oligosaccharid eller en stivelse; hvor sammensætningen, som omfatter en polysaccharid, oligosaccharid eller stivelse, eventuelt er en øl; (c) kombinere polypeptidet af trin (a) med sammensætningen af trin (b) under forhold, hvor polypeptidet kan hydrolysere polysacchariden, oligosacchariden eller stivelsen i sammensætningen, som anvendes til brygning eller produktion af alkohol.
17. Fremgangsmåde til fjernelse af blæk på papir eller fibre, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe en sammensætning omfattende papir eller fiber; og (c) bringe polypeptidet af trin (a) i kontakt med sammensætningen af trin (b) under forhold, hvor polypeptidet kan fjerne blæk fra papiret eller fiberen.
18. Fremgangsmåde til biologisk blegning af en sammensætning, omfattende: (') (a) tilvejebringe sammensætningen og polypeptidet af krav 6, eller et po-lypeptid, der kodes af nukleinsyren ifølge krav 1; og (a) behandle sammensætningen med polypeptidet af krav 6, eller det po-lypeptid, der kodes af nukleinsyren ifølge krav 1; eller (ii) fremgangsmåden af (i), hvor sammensætningen er et papir- eller pulpprodukt.
19. Fremgangsmåde til fremstilling af et brændstof, omfattende: (') (a) tilvejebringe polypeptidet ifølge krav 6, eller polypeptidet, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe en sammensætning omfattende polysaccharid, oligosaccha-rid eller stivelse; og (c) bringe polypeptidet af trin (a) i kontakt med sammensætningen af trin (b) under forhold, hvor polypeptidet hydrolyserer polysacchariden, oligosaccha-riden eller stivelsen; eller (ii) fremgangsmåden af (i), hvor polypeptidet er et termostabilt enzym; (iii) fremgangsmåden af (i), hvor brændstoffet er ethanol-baseret.
20. Fremgangsmåde til behandling af biomassemateriale, omfattende lig-nocellulose, omfattende (') (a) tilvejebringe en sammensætning, omfattende polypeptidet af krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1, og tilvejebringe et biomassemateriale; og (b) bringe sammensætningen, som omfatter polypeptidet af krav 6, eller polypeptidet, som kodes af nukleinsyren ifølge krav 1, i kontakt med biomassematerialet; (ii) fremgangsmåden af (i), hvor biomassematerialet omfatter eller er afledt af landbrugsafgrøder, eller er et biprodukt af en foder- eller fødevareproduktion, eller er et lignocellulosisk affaldsprodukt, eller er en planterest eller et affaldspapir eller affaldspapirproduktion, og biomassematerialet omfatter eventuelt kartofler, sojabønne, byg, rug, majs, havre, hvede, roer eller sukkerrør; (iii) fremgangsmåden af (i) eller (ii), hvor polypeptidet har aktivitet, som omfatter en glucoamylase-aktivitet; (iv) fremgangsmåde af (ii), hvor planteresten omfatter stammer, blade, bælge, skaller, kolber, træ, træsplinter, træmasse og savsmuld, eller papiraffaldet omfatter kasseret eller brugt fotokopipapir, computerprinterpapir, notes-bogpapir, notesblokpapir, skrivemaskinepapir, aviser, ugeblade, karton og papirbaseret indpakningsmaterialer; eller (v) fremgangsmåden af (i) til (iv), hvor behandlingen af biomassematerialet genererer bioethanol, biopropanol, biobutanol eller biodiesel; (vi) fremgangsmåde af (i) til (v), hvor biomassematerialet omfatter en lig-nocellulose.
21. Fremgangsmåde til fremstilling af sukker, hvor fremgangsmåden omfatter: (i) (a) tilvejebringe polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1; (b) tilvejebringe en polysaccharid eller oligosac-charid; og (c) bringe sammensætningen af trin (b) i kontakt med polypeptidet af trin (a), og derved generere sukker; (ii) fremgangsmåde af (i), hvor sammensætningen, som omfatter en polysaccharid, omfatter en stivelse; (iii) fremgangsmåden af en af (i) eller (ii), hvor sukkeret er et gæringsdygtigt sukker; (iv) fremgangsmåden af (i), (ii) eller (iii), yderligere omfattende gæring af sukkeret for at fremstille en alkohol; eller (v) fremgangsmåden af (iv), hvor alkoholen er ethanol, propanol eller butanol.
22. Detergentsammensætning, foder eller fødevare, væv eller tekstil, papir eller papirprodukt eller papirmasse, en væske med højt indhold af maltose eller glucose, en sammensætning til forhindring af brødtørring, en alkoholisk drik, en øl, en farmaceutisk sammensætning, et mundhygiejneprodukt, hvor mundhygiejneproduktet eventuelt omfatter en tandpasta, en dental crem, en gel eller et tandpulver, et tandplejemiddel, et mundskyllemiddel, en formulering til rensning før og efter børstning, tyggegummi, pastil eller bolsje, en olieboringsfluid, en biologisk blegningsopløsning, et desinficerende middel, et middel til biologisk forsvar eller biologisk afgiftning, eller et mælkeprodukt, eller et biomassemateriale, som omfatter polypeptidet ifølge krav 6, eller et polypeptid, der kodes af nukleinsyren ifølge krav 1.
23. Sammensætning med forsinket frigivelse eller kontrolleret frigivelse, omfattende polypeptid ifølge krav 6, eller et polypeptid, der kodes af nukleinsyre ifølge krav 1, omfattende (a) en ønsket ingrediens, coated med en latexpolymer-coating; (b) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse af (a), hvor det ønskede ingrediens omfatter et enzym; (c) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse af (a) eller (b), hvor den ønskede ingrediens omfatter et lille molekyle, et lægemiddel, en polysaccharid, en lipid, en nukleinsyre, et vitamin, antibiotika eller en insektgift; (d) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse af (a), (b) eller (c), hvor den ønskede ingrediens omfatter en pellet eller en matrix; (e) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse af en af (a) til (d), hvor pellet eller matrix omfatter spiseligt materiale; (f) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse af en (a) til (e), hvor latexpolymer-coatingen omfatter en latex-maling; eller (g) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse af en af (a) til (f), hvor latexpolymer-coatingen omfatter (meth)acrylat, vinylacetat, styren, ethylen, vinylchlorid, butadien, vinylidenchlorid, vinylversatat, vi-nylpropionat, t-butylacrylat, acrylonitril, neopren, maleat, fumarat eller en kombination deraf eller et derivat deraf; eller (h) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse af en af (a) til (g).
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DK12152656.0T DK2479266T3 (da) | 2006-12-21 | 2007-12-21 | Amylaser og glucoamylaser, nukleinsyrer, som koder for dem, og fremgangsmåder til fremstilling og anvendelse deraf |
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DK12152656.0T DK2479266T3 (da) | 2006-12-21 | 2007-12-21 | Amylaser og glucoamylaser, nukleinsyrer, som koder for dem, og fremgangsmåder til fremstilling og anvendelse deraf |
DK16180480.2T DK3101128T3 (da) | 2006-12-21 | 2007-12-21 | Amylaser og glucoamylaser, nukleinsyrer, der koder for disse, og fremgangsmåder til fremstillling og anvendelse deraf |
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