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DK2933329T3 - Hafniaphytasevarianter - Google Patents

Hafniaphytasevarianter Download PDF

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Publication number
DK2933329T3
DK2933329T3 DK15166896.9T DK15166896T DK2933329T3 DK 2933329 T3 DK2933329 T3 DK 2933329T3 DK 15166896 T DK15166896 T DK 15166896T DK 2933329 T3 DK2933329 T3 DK 2933329T3
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phytase
feed
seq
polypeptide
nucleic acid
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DK15166896.9T
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Soeren Flensted Lassen
Lars Kobberoee Skov
Maria Leonardo De
Tomoko Matsui
Allan Noergaard
Jesper Vind
Esben Peter Friis
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Novozymes As
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/16Hydrolases (3) acting on ester bonds (3.1)
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/189Enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/30Feeding-stuffs specially adapted for particular animals for swines
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/70Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75Feeding-stuffs specially adapted for particular animals for birds for poultry
    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02EREDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00Technologies for the production of fuel of non-fossil origin
    • Y02E50/10Biofuels, e.g. bio-diesel

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Birds (AREA)
  • Food Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Animal Husbandry (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Fodder In General (AREA)

Claims (19)

1. Phytasevariant med mindst 90% identitet med aminosyreresterne 1-413 i SEQ ID NO: 2, som har forbedrede egenskaber med hensyn til varmestabilitet sammenlignet med SEQ ID NO: 2, og som omfatter mindst en modifikation og ikke mere end 30 modifikationer i mindst en position valgt fra følgende: 1,5, 9, 12, 63, 69, 132, 138, A132V/Q181L, 186, 192, 207, A132V/E211R, 221,245, 251, 261,303, 348, 383 og 401, og hvor positionerne svarer til positionerne for phytasen med aminosyrerne 1-413 i SEQ ID NO: 2.
2. Phytasevariant ifølge krav 1 omfattende mindst en yderligere modifikation i mindst en position valgt fra følgende: 4, 6, 7, 8, 10, 16, 18, 25, 26, 27, 28, 29, 30, 31, 32, 33, 35, 36, 37, 38, 39, 40, 41,45, 48, 49, 54, 55, 59, 64, 66, 68, 69, 70, 71, 72, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 89, 91,92, 93, 95, 96, 97,98, 100, 101, 103, 108, 109, 110, 111, 112, 113, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 128, 130, 131, 133, 134, 136, 137, 139, 140, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 158, 159, 160, 161, 162, 163, 168, 172, 173, 175, 176, 177, 178, 179, 180, 182, 183, 184, 185, 187, 189, 190, 193, 194, 195, 196, 198, 199, 200, 201,202, 203, 204, 205, 206, 208, 209, 215, 217, 219, 224, 227, 228, 230, 233, 234, 235, 236, 238, 239, 240, 241,242, 243, 244, 246, 247, 248, 249, 256, 258, 259, 260, 266, 268, 270, 279, 284, 285, 286, 287, 288, 289, 290, 292, 293, 294, 295, 296, 297, 298, 299, 301, 304, 308, 310, 312, 313, 314, 316, 318, 319, 320, 322, 324, 325, 326, 331,335, 343, 344, 345, 346, 347, 354, 355, 356, 358, 360, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 378, 382, 384, 385, 386, 387, 388, 389, 390, 391,394, 395, 396, 397, 400, 403, 404, 406, 408, 409, 411, 412, og 413.
3. Phytasevariant ifølge et hvilket som helst af kravene 1 til 2, hvor modifikationen eller modifikationerne er valgt fra følgende: 1 P,Q, *1aG, *1aP, *1aS, *1bP, *1bS, *1cS, 1QGPS, 1QS, 9K,P,S, 12S,R, 63C, 69E,L, 132T,V, 138N,V, 186S,T, 192A, 207N,L,Q,R, 221T,G, 245E, 251S,D,E,R, 261F,A,Q, 303L, 348D,E,S,R, 383N og 401N.
4. Phytasevariant ifølge et hvilket som helst af kravene 1 til 3, som omfatter mindst en modifikation valgt fra følgende: 8C, 33C, 35C, 36C, 54C, 63C, 66C, 101 C, 139C, 143C, 201 C, 150C, 172C, 176C, 177C, 178C, 179C, 224C, 228C, 236C, 259C, 325C, 326C, 331 C, 358C, 363C, 368C, 370C, 374C.
5. Phytasevariant ifølge krav 4 omfattende mindst et sæt af modifikationer valgt fra følgende: 8C/343C, 1390/201 C, 33C/179C, 33C/178C, 35C/172C, 36C/177C, 36/C176C, 143C/201C, 540/101 C, 63C/368C, 66C/370C, 224C/236C, 150C/259C, 326C/331C, 3250/3580/, 228C/363C, 368C/374C.
6. Phytasevariant ifølge et hvilket som helst af kravene 1 til 5 omfattende mindst en modifikation valgt fra følgende: 29P, 30P, 93P, 95P, 140P, 163P, 235P, 243P, 244P, 284P, 287P, 288P, 316P, 360P, 8L, 16V, 27A, 30L, 32Q, 37D, 38A, 41T, 48W, 49L, 54G, 55E, 75A, 77K, 78G, 93E, 103A, 109D, 128N, 130L, 134V, 136Q, 137L, 173N, 176Q, 195L, 198E, 206T, 209S, 215S, 219M, 227Q, 228Q, 248T, 258Y, 260L, 31 OL, 313L, 314G, 316A, 318E, 319L, 320N, 335E, 354L, 356F, 360Q, 362M, 363R, 365K, 366T, 369S, 374P, 376E, 378K, 411S, 9R, 29R,K, 37R,K, 59R,K, 69E, 70E, 78R,K, 81E, 93E, 111 R,K, 115R,K, 119D, 185R.K, 230E, 239R, 293R,K, 363R,K, 395E, 396D,E, Q162N/D138N, A132V/Q181L, A132V/E211R, A132V/Q162R/Q181 L/A217G, P348R/H363R, Q9S/D92Y, A217G/K251S, E100W/A217G/K251S, E100W/K251S, Q9S/E100W/R160G/A217G/H363R, E100W/A217G/P348R/H363R, Q9S/A89A/D92Y/H115M/A217G/H363R, E100W/A217G/K234V/K251 E/I286T/H363R, E100W/A217G/K234V/P348R/H363R, Q9S/R18K/A89A/D92Y/H115M/A217G/K234V/H363R, Q9S/D92Y/H115M/A217G/K234V/H363 R, Q9S/N78Q/D92Y/L112S/H115M/K234V/P348R/H363R, Q9S/N78Q/A89A/D92Y/H115M/A132V/Q162R/Q181L/A217G/K234V/P348R, Q9S/E54C/D92Y/A101C/H143C/Q193R/I201C/A217G/H363R, Q9S/N78Q/D92Y/L112S/H115M/A217G/K234V/P348R/H363R, Q9S/N78Q/A89 A/D92Y/H115M/A132V/H143C/Q162R/Q181L/1201C/A217G/K234V/ P348R, Q9S/N78Q/A89 A/D92 Y/H115M/A132V/K139C/Q162R/Q181 L/1201 C/A217G/K234W P348R, Q9S/N78Q/A89A/D92Y/H115M/A132V/Q162R/Y179W/Q181L/A217G/K234V/P348R, Q9S/N78Q/A89A/D92Y/H115M/A132V/Q162R/Y179W/A217G/K234V/P348R/H363R, Q9S/N78Q/A89A/D92Y/H115M/A132V/Q162R/Y179W/A217G/K234V/S261F/P348R/ H363R, Q9S/N78Q/A89 A/D92Y/H115M/A132V/K139C/G151D/Q162R/Y179W/Q181 L/1201 Cl A217G/K234V/P348R, Q9S/N78Q/A132V/K139C/Q162R/Y179W/I201C/A217G/K234L7P348R/H363R, D92Y/E100W/K139C/I201C/A217G/K234V/N247D/Q256D/P348R/H363R, D92Y/E100W/K139C/Q162R/Q181 L/1201C/A217G/K234V/N247D/Q256D/H363R, Y48H/E54C/D92Y/A101C/K139C/T152A/I201C/K207Q/V208T/A217G/K234V/N247D/ R289W/H363R, Q162N/D138N, E54C/A101C/K207Q, E54C/A101C/K139C/I201C/K207Q, S1QGPS/K26H/E54C/A101C/K139C/Q162N/G186S/I201C/K207Q/G346S, Y48H/E54C/D92Y/A101C/K139C/T152A/I201C/K207Q/V208T/A217G/K234V/N247D/ R289W/H363R, og Q9S/N78Q/A89 A/D92Y/H115M/A132V/K139C/Q162R/Q181 L/1201 C/A217G/K234V/ P348R.
7. Fremgangsmåde til fremstilling af en phytasevariant ifølge et hvilket som helst af kravene 1 til 6 omfattende a) mutation af DNA’et eller genet, der koder for udgangsphytasen på en måde så DNA’et eller genet koder for substitutionen, insertionen og/eller deletionen, b) operabel forbindelse af DNA’et eller genet til en eller flere kontrolsekvenser, der styrer fremstillingen af phytasen i en passende ekspressionsvært til fremstilling af en DNA-konstruktion eller en rekombinant ekspressionsvektor, c) overføring af konstruktionen eller vektoren til en passende vært, d) kultivering af værten til fremstilling af phytasevarianten, og e) udvinding af phytasen.
8. Isoleret nukleinsyresekvens omfattende en nukleinsyresekvens, der koder for phytasevarianten ifølge et hvilket som helst af kravene 1 til 6.
9. Nukleinsyrekonstruktion omfattende nukleinsyresekvensen ifølge krav 8 operabelt forbundet med en eller flere kontrolsekvenser, der styrer fremstillingen af phytasen i en passende ekspressionsvært.
10. Rekombinant ekspressionsvektor omfattende nukleinsyrekonstruktionen ifølge krav 9.
11. Rekombinant værtscelle omfattende nukleinsyrekontruktionen ifølge krav 9 og/eller ekspressionsvektoren ifølge krav 10.
12. Fremgangsmåde til fremstilling af phytasevarianten ifølge et hvilket som helst af kravene 1 til 6 omfattende (a) kultivering af værtscellen ifølge krav 11 til fremstilling af en supernatant omfattende phytasen; og (b) udvinding af phytasen.
13. Sammensætning omfattende mindst en phytasevariant ifølge et hvilket som helst af kravene 1 til 6, og (a) mindst et fedtopløseligt vitamin; (b) mindst et vandopløseligt vitamin; og/eller; (c) mindst et spormineral.
14. Sammensætning ifølge krav 13 yderligere omfattende mindst et enzym valgt fra følgende gruppe af enzymer: amylase, phytase, phosphatase, xylanase, galactanase, alpha-galactosidase, protease, phospholipase, og/eller beta-glucanase.
15. Sammensætning ifølge et hvilket som helst af kravene 13 til 14, hvilken sammensætning er et dyrefoderadditiv.
16. Dyrefodersammensætning med et råproteinindhold på 50 til 800 g/kg og omfattende phytasevarianten ifølge et hvilket som helst af kravene 1 til 6 eller sammensætningen ifølge et hvilket som helst af kravene 13 til 15.
17. Fremgangsmåde til forbedring af næringsværdien af et dyrefoder, hvor phytasevarianten ifølge et hvilket som helst af kravene 1 til 6 eller sammensætningen ifølge et hvilket som helst af kravene 13 til 15 tilføjes til foderet.
18. Fremgangsmåde til reduktion af phytatniveaet i dyregødning omfattende fodring af dyret med en effektiv mængde af foderet ifølge krav 16.
19. Fremgangsmåde til fremstilling af et fermenteringsprodukt omfattende (a) fermentering af et materiale indeholdende kulhydrat under tilstedeværelse af en phytasevariant ifølge et hvilket som helst af kravene 1 til 6 og ved anvendelse af en fermenteringsmikroorganisme og (b) fremstilling af fermenteringsproduktet eller fermenteringsbiproduktet fra materialet indeholdende det fermenterede kulhydrat.
DK15166896.9T 2008-09-26 2009-09-28 Hafniaphytasevarianter DK2933329T3 (da)

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EP20130170321 EP2650364B1 (en) 2008-09-26 2009-09-28 Hafnia phytase variants

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US (2) US8206962B2 (da)
EP (3) EP2650364B1 (da)
CN (1) CN102165059A (da)
AU (1) AU2009295852B2 (da)
BR (1) BRPI0919314A2 (da)
CA (1) CA2736321C (da)
DK (3) DK2650364T3 (da)
ES (3) ES2426717T3 (da)
MX (1) MX2011003132A (da)
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