Xylose isomerases, nucleic acids encoding them and methods for making and using them

  • US 20060281908A1
  • Filed: 10/23/2003
  • Published: 12/14/2006
  • Est. Priority Date: 11/06/2002
  • Status: Abandoned Application
First Claim
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1. An isolated, synthetic or recombinant nucleic acid comprising:

  • (a) a nucleic acid sequence having at least 96%, 97%, 98%, 99%, or more, or complete (100%) sequence identity to SEQ ID NO;

    1 over a region of at least about 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150 or more residues, wherein the nucleic acid encodes a polypeptide having a xylose isomerase activity;

    or (a) nucleic acid sequence having at least 95%, 96%, 97%, 98%, 99%, or more, or complete (100%) sequence identity to SEQ ID NO;

    3 over a region of at least about 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150 or more residues, wherein the nucleic acid encodes a polypeptide having a xylose isomerase activity;

    or (c) a nucleic acid sequence encoding a polypeptide having a xylose isomerase activity comprising the sequence or SEQ ID NO;

    2, or SEQ ID NO;

    4, or enzymatically active fragments thereof;

    or (d) a nucleic acid sequence that hybridizes under stringent conditions to a nucleic acid comprising the sequence or SEQ ID NO;

    1, or SEQ ID NO;

    3, wherein the nucleic acid encodes a polypeptide having a xylose isomerase activity, wherein the sequence is at least about 25, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150 or more residues in length, or the full length of a gene or a transcript, wherein optionally the stringent conditions comprise a wash step comprising a wash in 0.2×

    SSC at a temperature of about 65°

    C. for about 15 minutes;

    or (e) a nucleic acid sequence completely complementary to (a), (b), (c) or (d), wherein the nucleic acid encodes at least one polypeptide having a xylose isomerase activity, and optionally the sequence identities are determined by analysis with a sequence comparison algorithm or by a visual inspection, and optionally the sequence comparison algorithm is a BLAST version 2.2.2 algorithm where a filtering setting is set to blastall -p blastp -d “

    nr pataa”

    -F F, and all other options are set to default, and optionally the xylose isomerase activity comprises;

    isomerization of xylose to xylulose;

    isomerization of glucose to fructose;

    isomerization of D-glucose to D-fructose;

    catalysis of the conversion of D-xylose to an equilibrium mixture of D-xylulose and D-xylose;

    isomerization of α

    -D-glucopyranose to α

    -D-fructofuranose;

    isomerization of β

    -D-glucopyranose to β

    -D-fructopyranose, and optionally the xylose isomerase activity is thermostable, and optionally the polypeptide retains a xylose isomerase activity under conditions comprising a temperature range of between about 60°

    C. to about 120°

    C., or, between about 60°

    C. to about 95°

    C., or retains a xylose isomerase activity under conditions comprising a temperature range of between about 95°

    C. to about 105°

    C., or, between about 105°

    C. to about 120°

    C., and optionally the xylose isomerase activity thermotolerant, and optionally retains a xylose isomerase activity after exposure to conditions comprising a temperature range of between about 95°

    C. to about 135°

    C., or, between about 95°

    C. to about 105°

    C., or between about 105°

    C. to about 120°

    C., or, between about 120°

    C. to about 135°

    C.

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