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    2-O-sulfation enzyme mutant and 3-O-sulfation enzyme mutant, and method for using same
    2.
    发明授权
    2-O-sulfation enzyme mutant and 3-O-sulfation enzyme mutant, and method for using same 有权

    公开(公告)号：US12139728B2

    公开(公告)日：2024-11-12

    申请号：US18056853

    申请日：2022-11-18

    Applicant: AJINOMOTO CO., INC.

    Inventor： Chihiro Tsuji ,  Tomoko Shimizu ,  Uno Tagami ,  Yasuhiro Mihara ,  Masayuki Sugiki ,  Shogo Nakano ,  Tomoharu Motoyama ,  Sohei Ito

    IPC: C12N9/10 ,  C08B37/00 ,  C12P19/26

    Abstract: The present invention provides a 2-OST mutant exhibiting a high activity. Specifically, the present invention provides a 2-O-sulfation enzyme mutant, having a substitution of a leucine residue at position 321 with a basic amino acid residue in any one amino acid sequence of: (a) the amino acid sequence of SEQ ID NO: 2; (b) an amino acid sequence comprising one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence of SEQ ID NO: 2; (c) an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (d) the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; (e) an amino acid sequence comprising one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; (f) an amino acid sequence having 90% or more identity to the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; and having a 2-O-sulfate transfer activity.

    2-O-sulfation enzyme mutant and 3-O-sulfation enzyme mutant, and method for using same
    3.
    发明授权
    2-O-sulfation enzyme mutant and 3-O-sulfation enzyme mutant, and method for using same 有权

    公开(公告)号：US11851688B2

    公开(公告)日：2023-12-26

    申请号：US16795991

    申请日：2020-02-20

    Applicant: AJINOMOTO CO., INC.

    Inventor： Chihiro Tsuji ,  Tomoko Shimizu ,  Uno Tagami ,  Yasuhiro Mihara ,  Masayuki Sugiki ,  Shogo Nakano ,  Tomoharu Motoyama ,  Sohei Ito

    IPC: C12N9/10 ,  C08B37/00 ,  C12P19/26

    CPC classification number: C12N9/13 ,  C08B37/0075 ,  C12P19/26 ,  C12Y208/02

    Abstract: The present invention provides a 2-OST mutant exhibiting a high activity. Specifically, the present invention provides a 2-O-sulfation enzyme mutant, having a substitution of a leucine residue at position 321 with a basic amino acid residue in any one amino acid sequence of: (a) the amino acid sequence of SEQ ID NO: 2; (b) an amino acid sequence comprising one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence of SEQ ID NO: 2; (c) an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (d) the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; (e) an amino acid sequence comprising one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; (f) an amino acid sequence having 90% or more identity to the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; and having a 2-O-sulfate transfer activity.

    Heparosan-Producing Bacterium and Heparosan Manufacturing Method
    4.
    发明申请
    Heparosan-Producing Bacterium and Heparosan Manufacturing Method 审中-公开

    公开(公告)号：US20180237479A1

    公开(公告)日：2018-08-23

    申请号：US15958354

    申请日：2018-04-20

    Applicant: AJINOMOTO CO., INC.

    Inventor： Shunsuke Yamazaki ,  Tomoko Shimizu ,  Kenichi Mori ,  Naoto Tonouchi

    IPC: C07K14/245 ,  C12P19/04 ,  C08B37/00 ,  C12N15/70 ,  C12P19/26

    Abstract: A method for producing heparosan is provided. Heparosan is produced by culturing an Escherichia bacterium having a heparosan-producing ability and modified so that expression of one or more genes, such as rbsR, rbsK, rbsB, hsrA, glgB, glgX, micF, rcsD, rcsB, ybiX, ybil, ybiJ, ybiC, ybiB, rfaH, nusG, pcoR, pcoS, pcoE, yhcN, yhcO, aaeB, aaeA, aaeX, g1455, alpA, g1453, yrbA, mlaB, mlaC, mlaD, mlaE, mlaF, yrbG, norW, ybjI, ybjJ, ybjK, rybB, yjjY, yjtD, thrL, thrA, thrB, fruA, psuK, ytfT, yjfF, fbp, yagU, paoA, paoB, gsiC, gsiD, yliE, irp2, irp1, bhsA, ycfS, lepB, rnc, era, dapA, gcvR, bcp, hyfA, rpoE, nadB, yfiC, srmB, g1414, g1413, nuoE, nuoF, nuoG, glmZ, hemY, hemX, hemD, rlmL, artQ, artM, artJ, rlmC, ybjO, yejO, yejM, yejL, rpoS, ygbN, ygbM, ygbL, g3798, g3797, g3796, g3795, g3794, g3793, g3792, ryjA, soxR, soxS, yjcC, yjcB, efeU, and efeO, is/are increased in a medium, and collecting heparosan from the medium.

    2-O-Sulfation Enzyme Mutant and 3-O-Sulfation Enzyme Mutant, and Method for Using Same
    7.
    发明申请
    2-O-Sulfation Enzyme Mutant and 3-O-Sulfation Enzyme Mutant, and Method for Using Same 审中-公开

    公开(公告)号：US20200181588A1

    公开(公告)日：2020-06-11

    申请号：US16795991

    申请日：2020-02-20

    Applicant: AJINOMOTO CO., INC.

    Inventor： Chihiro Tsuji ,  Tomoko Shimizu ,  Uno Tagami ,  Yasuhiro Mihara ,  Masayuki Sugiki ,  Shogo Nakano ,  Tomoharu Motoyama ,  Sohei Ito

    IPC: C12N9/10 ,  C12P19/26 ,  C08B37/00

    Abstract: The present invention provides a 2-OST mutant exhibiting a high activity. Specifically, the present invention provides a 2-O-sulfation enzyme mutant, having a substitution of a leucine residue at position 321 with a basic amino acid residue in any one amino acid sequence of: (a) the amino acid sequence of SEQ ID NO: 2; (b) an amino acid sequence comprising one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence of SEQ ID NO: 2; (c) an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (d) the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; (e) an amino acid sequence comprising one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; (f) an amino acid sequence having 90% or more identity to the amino acid sequence consisting of amino acid residues at positions 69 to 356 in the amino acid sequence of SEQ ID NO: 2; and having a 2-O-sulfate transfer activity.

    Heparosan-Producing Bacterium and Heparosan Manufacturing Method
    8.
    发明申请
    Heparosan-Producing Bacterium and Heparosan Manufacturing Method 有权
    Heparosan生产细菌和Heparosan制造方法

    公开(公告)号：US20160201103A1

    公开(公告)日：2016-07-14

    申请号：US15082464

    申请日：2016-03-28

    Applicant: AJINOMOTO CO., INC.

    Inventor： Shunsuke Yamazaki ,  Tomoko Shimizu ,  Kenichi Mori ,  Naoto Tonouchi

    IPC: C12P19/04 ,  C12N15/70

    CPC classification number: C07K14/245 ,  C08B37/0075 ,  C12N15/70 ,  C12P19/04 ,  C12P19/26

    Abstract: A method for producing heparosan is provided. Heparosan is produced by culturing an Escherichia bacterium having a heparosan-producing ability and modified so that expression of one or more genes, such as rbsR, rbsK, rbsB, hsrA, glgB, glgX, micF, rcsD, rcsB, ybiX, ybiI, ybiJ, ybiC, ybiB, rfaH, nusG, pcoR, pcoS, pcoE, yhcN, yhcO, aaeB, aaeA, aaeX, g1455, alpA, g1453, yrbA, mlaB, mlaC, mlaD, mlaE, mlaF, yrbG, norW, ybjI, ybjJ, ybjK, rybB, yjjY, yjtD, thrL, thrA, thrB, fruA, psuK, ytfT, yjfF, fbp, yagU, paoA, paoB, gsiC, gsiD, yliE, irp2, irp1, bhsA, ycfS, lepB, rnc, era, dapA, gcvR, bcp, hyfA, rpoE, nadB, yfiC, srmB, g1414, g1413, nuoE, nuoF, nuoG, glmZ, hemY, hemX, hemD, rlmL, artQ, artM, artJ, rlmC, ybjO, yejO, yejM, yejL, rpoS, ygbN, ygbM, ygbL, g3798, g3797, g3796, g3795, g3794, g3793, g3792, ryjA, soxR, soxS, yjcC, yjcB, efeU, and efeO, is/are increased in a medium, and collecting heparosan from the medium.

    Abstract translation: 提供了一种生产heparosan的方法。 Heparosan是通过培养具有肝素生产能力的埃希氏菌属细菌产生的，并被修饰，使得一种或多种基因如rbsR，rbsK，rbsB，hsrA，glgB，glgX，micF，rcsD，rcsB，ybiX，ybiI，ybiJ ，ybiC，ybiB，rfaH，nusG，pcoR，pcoS，pcoE，yhcN，yhcO，aaeB，aaeA，aaeX，g1455，alpA，g1453，yrbA，mlaB，mlaC，mlaD，mlaE，mlaF，yrbG，norW，ybjI，ybjJ ，ybjK，rybB，yjjY，yjtD，thrL，thrA，thrB，fruA，psuK，ytfT，yjfF，fbp，yagU，paoA，paoB，gsiC，gsiD，yliE，irp2，irp1，bhsA，ycfS，lepB， ，dapA，gcvR，bcp，hyfA，rpoE，nadB，yfiC，srmB，g1414，g1413，nuoE，nuoF，nuoG，glmZ，hemY，hemX，hemD，rlmL，artQ，artM，artJ，rlmC，ybjO，yejO，yejM ，在培养基中增加yejL，rpoS，ygbN，ygbM，ygbL，g3798，g3797，g3796，g3795，g3794，g3793，g3792，ryjA，soxR，soxS，yjcC，yjcB，efeU和efeO， heparosan从媒介。

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