Hydrolases, nucleic acids encoding them and methods for making and using them

Hydrolases, nucleic acids encoding them and methods for making and using them

  • CN 102,197,043 A
  • Filed: 08/28/2009
  • Published: 09/21/2011
  • Est. Priority Date: 08/29/2008
  • Status: Active Grant
First Claim
Patent Images

1. the nucleic acid of isolating, synthetic or reorganization, it comprises(a), the encode nucleic acid (polynucleotide) of at least a polypeptide, wherein said nucleic acid comprises with following having at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or it is bigger, or the sequence of (100%) sequence identity completely:

  • (i) SEQ ID NO;

    1, SEQ ID NO;

    3, SEQ ID NO;

    5, SEQ ID NO;

    7, SEQ ID NO;

    9, SEQ ID NO;

    11, SEQ ID NO;

    13, SEQ ID NO;

    15, SEQ ID NO;

    17, SEQ ID NO;

    19, SEQ ID NO;

    22 or SEQ ID NO;

    23, orThe (ii) nucleic acid of SEQ ID NO;

    1, it has one of coding, two, three, four, five, six, seven, eight, nine, ten, 11,12,13,14,15,16,17,18, nineteen, 20,21,22,23,24 or more a plurality of or all as table 3, table 4, table 9, table 10, table 11, one or more Nucleotide that amino acid shown in table 16 or the table 23 changes (or its equivalent variations) change (or its equivalent variations)At least a polypeptide of wherein said (i) or nucleic acid encoding (ii) with hydrolytic enzyme activities, perhaps coding can produce the polypeptide or the peptide (serving as epi-position or immunogenic polypeptide or peptide) of lytic enzyme specific antibody,(b), (a) nucleic acid (polynucleotide), the following mensuration of wherein said sequence identity;

    (A) by analyzing with sequence comparison algorithm or passing through visual inspection, or (B) at least about 10,15,20,25,30,35,40,45,50,55,60,65,70,75,100,125,150,175,200,250,300,350,400,450,500,550,600,650,700,750,800,850,900,950,1000,1050,1100,1150,1200,1250,1300,1350,1400,1450,1500,1550 or the zone of more a plurality of residues in, or at cDNA, in the total length zone of transcript (mRNA) or gene(c), (a) or nucleic acid (b) (polypeptide), wherein said sequence comparison algorithm is a BLAST 2.2.2 version algorithm, wherein filter to be provided with to be set to blastall-p blastp-d " nr pataa "-F F, and all other options are set to default value,(d), nucleic acid (polynucleotide), its at least a polypeptide or polypeptide of encoding with hydrolytic enzyme activities, wherein said nucleic acid is included under the stringent condition sequence with the complementary sequence hybridization of (a) and (b) or nucleic acid (c), wherein said stringent condition is included in and washs about 15 minutes washing step under about 65 ℃

    temperature in 0.2 * SSC(e), nucleic acid (polynucleotide), its at least a for example lipase of hydrolytic enzyme activities that has of encoding, saturated enzyme, the active polypeptide of palmitinic acid enzyme and/or stearic acid enzymes, wherein said polypeptide comprises sequence or its enzymatic activity fragment of SEQ ID NO;

    2, has at least one, two, three, four, five, six, seven, eight, nine, ten, 11,12,13,14,15,16,17,18, nineteen, 20,21,22,23,24 or more a plurality of or all as table 3, table 4, table 9, table 10, table 11, amino acid shown in table 16 or the table 23 changes (or its equivalent variations)(f), nucleic acid (polynucleotide), its at least a polypeptide of encoding with hydrolytic enzyme activities, wherein said polypeptide comprises sequence or its enzymatic activity fragment of SEQ ID NO;

    2, SEQ ID NO;

    4, SEQ ID NO;

    6, SEQ ID NO;

    8, SEQ ID NO;

    10, SEQ ID NO;

    12, SEQ ID NO;

    14, SEQ ID NO;

    16, SEQ ID NO;

    18 or SEQ ID NO;

    20(g), (A), (a) to (f) each and nucleic acid encoding (polynucleotide), described polypeptide has at least one conservative amino acid displacement and keeps its hydrolytic enzyme activities, or (B), (g) nucleic acid (A), wherein said described at least one conservative amino acid displacement comprises the amino-acid substitution amino acid with another similar characteristics;

    Perhaps, preservative replacement comprises;

    replace aliphatic amino acid with another aliphatic amino acid;

    Replace Serine or vice versa with Threonine;

    Replace acidic residues with another acidic residues;

    Replace the residue that contains amide group with another residue that contains amide group;

    With another alkaline residue exchange alkaline residue;

    Or replace aromatic residue with another aromatic residue,(h), (a) to (g) each, nucleic acid encoding (polynucleotide), described polypeptide has hydrolytic enzyme activities but lacks signal sequence,(i), (a) to (h) each, nucleic acid encoding (polynucleotide), described polypeptide has hydrolytic enzyme activities, further comprises heterologous sequence,(j), nucleic acid (i) (polynucleotide), wherein said described heterologous sequence comprise the following sequence of coding or are made up of the following sequence of coding;

    (A) allos signal sequence;

    (B), sequence (A), wherein said allos signal sequence derives from isodynamic enzyme;

    Perhaps (C) but mark, epi-position, target peptide, can cut sequence test section or enzyme, or(k), nucleotide sequence (polynucleotide), its with (a) to (j) each fully (fully) complementation of sequence.

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