DK2479266T3 - Amylaser og glucoamylaser, nukleinsyrer, som koder for dem, og fremgangsmåder til fremstilling og anvendelse deraf - Google Patents

Amylaser og glucoamylaser, nukleinsyrer, som koder for dem, og fremgangsmåder til fremstilling og anvendelse deraf Download PDF

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DK2479266T3
DK2479266T3 DK12152656.0T DK12152656T DK2479266T3 DK 2479266 T3 DK2479266 T3 DK 2479266T3 DK 12152656 T DK12152656 T DK 12152656T DK 2479266 T3 DK2479266 T3 DK 2479266T3
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polypeptide
nucleic acid
composition
sequence
starch
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Malgorzata Slupska
Yan Zhang
Geoff Hazlewood
Cathy Chang
Peter Luginbuhl
Ellen Burke
Michelle Cayouette
Uvini Gunavardena
Majid Ghassemian
Aron Silverstone
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Syngenta Participations Ag
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    • C12N9/2408Glucanases acting on alpha -1,4-glucosidic bonds
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    • C12N9/2405Glucanases
    • C12N9/2408Glucanases acting on alpha -1,4-glucosidic bonds
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    • C12N9/2405Glucanases
    • C12N9/2408Glucanases acting on alpha -1,4-glucosidic bonds
    • C12N9/2411Amylases
    • C12N9/2428Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
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    • Y02E50/00Technologies for the production of fuel of non-fossil origin
    • Y02E50/10Biofuels, e.g. bio-diesel

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Claims (26)

1. Isoleret, syntetisk eller rekombinant nukleinsyre bestående af: (a) en nukleinsyresekvens, der er mindst 98%, 99% eller 100% identisk med SEQ ID NO:51, hvor nukleinsyren koder for mindst et polypeptid med en alpha-amylase-aktivitet; (b) en nukleinsyresekvens, der koder for et polypeptid med sekvensen i SEQ ID NO:52; (c) en nukleinsyresekvens, der er fuldstændig komplementær til (a) eller (b); eller (d) en nukleinsyresekvens som angivet i SEQ ID NO:51.
2. Isoleret, syntetisk eller rekombinant nukleinsyresekvens som angivet i SEQ ID NO:51, hvor nukleinsyren koder for et polypeptid med en alpha-amylase-aktivitet og en aminosyresekvens som angivet i SEQ ID NO:52.
3. Vektor, ekspressionskassette eller kloningsvehikel omfattende en nukleinsyre, der omfatter nukleinsyresekvensen ifølge krav 1; hvor nukleinsyren eventuelt er funktionsmæssigt forbundet med en promoter; hvor promoteren eventuelt er en plantepromotor eller en viral promotor; hvor hvor promotoren eventuelt er en inducerbar promotor eller en vævsspecifik promotor; 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 nukleinsyresekvensen ifølge krav 1, eller en vektor, ekspressionskassette eller kloningsvehikel ifølge krav 3, hvor cellen eventuelt er en bakteriecelle, en pattedyrscelle, en svampecelle, en gærcelle, en insektcelle eller en plantecelle, hvor plantecellen eventuelt er en kartoffel-, ris-, majs-, hvede-, tobaks- eller bygcelle.
5. Isoleret, syntetisk eller rekombinant polypeptid med alpha-amylase-aktivitet, omfattende: (i) en aminosyresekvens, der er mindst 99 % eller fuldstændig identisk med sekvensen med SEQ ID NO:52 over den fulde længde af polypeptidet; (ii) en aminosyresekvens, der kodes af nukleinsyren ifølge krav 1; (iii) polypeptidet ifølge et hvilket som helst af (i) til (ii), hvor polypeptidet har en sekvens som angivet i SEQ ID NO:52, eller enzymatisk aktive fragmenter deraf med alpha-amylase acitivity.
6. Isoleret, syntetisk eller rekombinant nukleinsyre ifølge krav 5, hvor alpha-amylase-aktiviteten omfatter hydrolysering af glucosidiske bindinger, og de glucosidiske bindinger eventuelt omfatter en a-1,4-glucosidisk binding eller en a-1,6-glucosidisk binding, og/eller hvor alpha-amylase-aktiviteten eventuelt omfatter hydrolysering af glucosidiske bindinger i en polysaccharid, en oligosaccharid eller en stivelse, og/eller alpha-amylase-aktiviteten omfatter spaltning af en maltose eller en D-glucose-enhed fra ikke-reducerende ende af en polysaccharid, oligosaccharid eller stivelse.
7. Fremgangsmåde til fremstilling af et rekombinant polypeptid, omfattende trinnene: (a) tilvejebringe en nukleinsyre, der er funktionsmæssigt forbundet med en promoter, hvor nukleinsyren omfatter nukleinsyresekvensen ifølge krav 1; og (b) eksprimere nukleinsyre fra trin (a) under betingelser, der muliggør eksprimering af polypeptidet, hvorved der fremstilles et rekombinant polypeptid, yderligere omfattende at transformere en værtscelle med nukleinsyren fra trin (a) efterfulgt af eksprimering af nukleinsyren fra trin (a), hvorved der fremstilles et rekombinant polypeptid i en transformeret celle.
8. Fremgangsmåde til at hydrolysere en polysaccharid, oligosaccharid eller stivelse, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 5; (b) tilvejebringe en sammensætning omfattende en polysaccharid, oligosaccharid eller stivelse; hvor sammensætningen eventuelt omfatter en a-1,4-glucosidisk binding eller en a-1,6-glucosidisk binding; og (c) bringe polypeptidet fra trin (a) i kontakt med sammensætningen fra trin (b) under betingelser, hvor polypeptidet hydrolyserer polysacchariden, oligosac-chariden eller stivelsen.
9. Fremgangsmåde til at fjerne eller gøre en polysaccharid, oligosaccharid eller stivelse flydende, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 5; (b) tilvejebringe en sammensætning omfattende en polysaccharid, oligosaccharid eller stivelse; og (c) bringe polypeptidet fra trin (a) i kontakt med sammensætningen fra trin (b) under betingelser, hvor polypeptidet fjerner eller gør polysacchariden, olig-osacchariden eller stivelsen flydende.
10. Fremgangsmåde til at vaske en genstand, omfattende følgende trin: (a) tilvejebringe en sammensætning omfattende polypeptidet ifølge krav 5; (b) tilvejebringe en genstand; og (c) bringe bringe polypeptidet fra trin (a) i kontakt med genstanden fra trin (b) under betingelser, hvor sammensætningen kan vaske genstanden.
11. 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 5; og (b) tilsætte polypeptidet ifølge krav 5 til foder- eller fødevarematerialet i en tilstrækkelig mængde 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 i foderet eller fødevaren inden fortæring af dyret, hvor foderet eller fødevaren eventuelt omfatter ris, majs, byg, hvede, grøntsager eller kartoffel.
12. Fremgangsmåde til krympning af tekstiler, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 5; (b) tilvejebringe stof; og (c) bringe polypeptidet fra trin (a) i kontakt med stoffet fra trin (b) under betingelser, hvor alpha-amylasen kan krympe stoffet.
13. Fremgangsmåde til behandling af lignocellulosiske fibre, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 5; (b) tilvejebringe en lignocellulosisk fiber; og (c) bringe polypeptidet fra trin (a) i kontakt med fiberen fra trin (b) under betingelser, hvor polypeptidet kan behandle fiberen og derved forbedre fiberegenskaberne.
14. Fremgangsmåde til at fremstille en sirup med højt maltoseindhold eller glucoseindhold, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 5; (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 fra trin (a) i kontakt med sammensætningen fra trin (b) under betingelser, hvor polypeptidet fra trin (a) kan hydrolysere sammensætningen fra trin (b), og derved fremstille en sirup med højt maltoseindhold eller glucoseindhold.
15. Fremgangsmåde til forbedring af strømmen af polysaccharid-, oligosac-charid- eller stivelsesholdige produktionsfluider, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 5; (b) tilvejebringe produktionsfluid omfattende en polysaccharid, oligosaccharid eller stivelse; hvor produktionsfluiden eventuelt er fra en underjordisk formation; og (c) bringe polypeptidet fra trin (a) i kontakt med produktionsfluiden fra trin (b) under betingelser, hvor polypeptidet kan hydrolysere polysacchariden, oligosacchariden eller stivelsen i produktionsfluiden og derved forbedre strømmen deraf ved at forøge densiteten.
16. Fremgangsmåde til ændring af viskositeten af en sammensætning, omfattende følgende trin: (i) (a) tilvejebringe en sammensætning og polypeptidet ifølge krav 5; og (b) behandle sammensætningen med polypeptidet ifølge krav 5; eller (ii) fremgangsmåden fra (i), hvor sammensætningen omfatter jord eller boreslam.
17. Fremgangsmåde til anvendelse af en alpha-amylase i brygning eller fremstilling af alkohol, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 5; (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 fra trin (a) med sammensætningen fra trin (b) under betingelser, hvor polypeptidet kan hydrolysere polysacchariden, olig-osacchariden eller stivelsen i sammensætningen, som anvendes til brygning eller produktion af alkohol.
18. Fremgangsmåde til biologisk blegning af en sammensætning, omfattende: (i) tilvejebringe en sammensætning, hvor sammensætningen er et papir- eller pulpprodukt, og polypeptidet ifølge krav 5; (ii) behandle sammensætningen med polypeptidet ifølge krav 5.
19. Fremgangsmåde til fjernelse af blæk på papir og fibre, omfattende følgende trin: (a) tilvejebringe polypeptidet ifølge krav 5; (b) tilvejebringe en sammensætning omfattende papir eller fiber; (c) bringe polypeptidet fra trin (a) i kontakt med sammensætningen fra trin (b) under betingelser, hvor polypeptidet kan fjerne blæk fra papiret eller fiberen.
20. Fremgangsmåde til fremstilling af et brændstof, omfattende: (i) (a) tilvejebringe polypeptidet ifølge krav 5; (b) tilvejebringe en sammensætning omfattende en polysaccharid, oligosaccharid eller stivelse; og (c) bringe polypeptidet fra (a) i kontakt med sammensætningen fra (b) under betingelser, hvor polypeptidet hydrolyserer polysacchariden, oligosacchariden eller stivelsen; eller (ii) fremgangsmåden fra (i), hvor polypeptidet er et termostabilt enzym; (iii) fremgangsmåden fra (i), hvor brændstoffet er ethanol-baseret.
21. Fremgangsmåde til behandling af et biomassemateriale omfattende lig-nocellulose, omfattende (i) (a) tilvejebringe en sammensætning, omfattende polypeptidet fra krav 5, og tilvejebringe et biomassemateriale; og (b) bringe sammensætningen i kontakt med biomassematerialet; (ii) fremgangsmåden fra (i), hvor biomassmaterialet omfatter eller er afledt af en landbrugsafgrøde, eller er et biprodukt af en foder- eller fødevareproduktion, eller er et lignocellulosisk affaldsprodukt, eller er en planterest eller et affaldspapir eller affaldspapirprodukt, og biomassematerialet eventuelt omfatter kartofler, sojabønne, byg, rug, majs, havre, hvede, roer eller sukkerrør; (iii) fremgangsmåden fra (i) eller (ii), hvor polypeptidet har aktivitet, som omfatter en alpha-amylase-aktivitet; (iv) fremgangsmåden fra (ii), hvor planteresten omfatter stammer, blade, bælge, skaller, kolber, træ, træsplinter, træmasse og savsmuld, eller papiraffaldet omfatter kasseret eller brugt fotokopipapir, computerprinterpapir, no-tesbogpapir, notesblokpapir, skrivemaskinepapir, aviser, ugeblade, karton og papirbaserede indpakningsmaterialer; eller (v) fremgangsmåden fra (i) til (iv), hvor behandlingen af biomassematerialet genererer en bioethanol, biopropanol, biobutanol eller en biodiesel; (vi) fremgangsmåden fra (i) til (v), hvor biomassematerialet omfatter en lig-nocellulose.
22. Fremgangsmåde til fremstilling af bioethanol, biopropanol, biobutanol eller en biodiesel, omfattende (i) (a) tilvejebringe polypeptidet ifølge krav 5, og tilvejebringe en sammensætning omfattende en polysaccharid eller oligosaccharid; og (b) bringe sammensætningen omfattende en polysaccharid eller oligosaccharid i kontakt med polypeptidet ifølge krav 5; (ii) fremgangsmåden fra (i), hvor sammensætningen omfattende en polysaccharid eller oligosaccharid omfatter en plante, et planteprodukt eller plantede-ri vat; (iii) fremgangsmåden fra (ii), hvor planten eller planteproduktet omfatter rørsukkerplanter eller -planteprodukter, roer eller sukkerroer, hvede, majs, sojabønner, kartoffel, ris eller byg; (iv) fremgangsmåden fra en hvilken som helst af (i) til (iii), hvor polypeptidet har aktivitet omfattende en alpha-amylase-aktivitet; eller (v) fremgangsmåden fra en hvilken som helst af (i) til (iv), hvor polysacchari-den eller oligosacchariden omfatter et fermenterbart sukker.
23. Fremgangsmåde til fremstilling af et brændstof, omfattende (i) (a) tilvejebringe polypeptidet ifølge krav 5 og tilvejebringe en sammensætning omfattende et fermenterbart sukker; og (b) bringe sammensætningen omfattende et fermenterbart sukker i kontakt med polypeptider ifølge krav 5; (ii) fremgangsmåden fra (i), hvor sammensætningen omfattende et fermenterbart sukker omfatter en plante, et planteprodukt eller plantederivat; (iii) fremgangsmåden fra (ii), hvor planten eller planteproduktet omfatter rørsukkerplanter eller -planteprodukter, roer eller sukkerroer, hvede, majs, sojabønner, kartoffel, ris eller byg; (iv) fremgangsmåde fra en hvilken som helst af (i) til (iii), hvor polypeptidet har aktivitet omfattende en alpha-amylase-aktivitet; eller (v) fremgangsmåde fra en hvilken som helst af (i) til (iv), hvor brændstoffet omfatter en bioethanol eller en benzin-ethanol-blanding eller omfatter en bio-ethanol, biopropanol, biobutanol eller en biodiesel.
24. Fremgangsmåde til fremstilling af sukker, hvilken fremgangsmåde omfatter: (i) (a) tilvejebringe polypeptidet ifølge krav 5; (b) tilvejebringe en sammensætning omfattende en polysaccharid eller en oligosaccharid; og (c) bringe sammensætningen fra trin (b) i kontakt med polypeptidet fra trin (a) og derved generere sukker; (ii) fremgangsmåden fra (i), hvor sammensætningen, som omfatter en polysaccharid, omfatter en stivelse; (iii) fremgangsmåden fra en hvilken som helst af (i) eller (ii), hvor sukkeret er et fermenterbart sukker.
25. Sammensætning omfattende polypeptidet ifølge krav 5, hvor sammensætningen er udvalgt blandt: et detergent, et foder eller en fødevare, tekstil eller stof, papir eller papirprodukt eller papirmasse, væske eller sirup med højt indhold af maltose eller glucose, en sammensætning til forhindring af brødtørring, en alkoholisk drik, en øl, en farmaceutisk sammensætning, et mundhygiejneprodukt, en tandpasta, en tandcreme, en gel eller et tandpulver, et tandplejemiddel, et mundskyllemiddel, en formulering til rensning før og efter børstning, tyggegummi, en pastil eller et bolsje, en olieboringsfluid, en biologisk blegningsopløsning, et desinficerende middel, et middel til biologisk forsvar eller biologisk afgiftning, et mejeriprodukt eller et biomassemateriale.
26. Sammensætning med forsinket frigivelse eller kontrolleret frigivelse omfattende polypeptidet ifølge krav 5, omfattende (a) en ønsket ingrediens, coatet med en latexpolymer-coating; (b) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse fra (a), hvor den ønskede ingrediens omfatter et enzym; (c) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse fra (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 fra (a), (b) eller (c), hvor den ønskede ingrediens omfatter en pellet eller en matrix; (e) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse fra en hvilken som helst af (a) til (d), hvor pellet eller matrix omfatter spiseligt materiale; (f) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse fra en hvilken som helst af (a) til (e), hvor latexpolymer-coatingen omfatter en latex-maling; eller (g) sammensætningen med forsinket frigivelse eller kontrolleret frigivelse fra en hvilken som helst af (a) til (f), hvor latexpolymer-coatingen omfatter (meth)acrylat, vinylacetat, styren, ethylen, vinylchlorid, butadien, vinyliden-chlorid, vinylversatat, vinylpropionat, t-butylacrylat, acrylonitril, neopren, ma-leat, fumarat eller en kombination deraf eller et derivat deraf.
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