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公开(公告)号：US11639513B2
公开(公告)日：2023-05-02
申请号：US16515415
申请日：2019-07-18
Applicant: AJINOMOTO CO., INC.
Inventor： Chihiro Tsuji , Yasuhiro Mihara , Shogo Nakano , Tomoharu Motoyama , Souhei Ito
Abstract: The present invention provides a method for producing a heparosan compound having an isomerized hexuronic acid residue and a method for producing a heparan sulfate with improved C5-epimerization efficiency. Specifically, the present invention provides a method for producing a heparosan compound having an isomerized hexuronic acid residue, said method comprising producing the heparosan compound having the isomerized hexuronic acid residue from a heparosan compound in the presence of a protein selected from the group of consisting of the following (A) to (F): (A) a protein comprising the amino acid sequence of SEQ ID No:2; (B) a protein which comprises an amino acid sequence having 90% or more homology to the amino acid sequence of SEQ ID No:2 and has a D-glucuronyl C5-epimerase substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID No:2 and has a D-glucuronyl C5-epimerase activity; (D) a protein comprising the amino acid sequence of SEQ ID NO:5; (E) a protein which comprises an amino acid sequence having 90% or more homology to the amino acid sequence of SEQ ID No:5 and has a D-glucuronyl C5-epimerase activity; and (F) a protein which comprises an amino acid sequence having one or several deleted, substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID No:5 and has a D-glucuronyl C5-epimerase activity.
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Patent Agency Ranking
公开(公告)号：US20190338330A1
公开(公告)日：2019-11-07
申请号：US16515415
申请日：2019-07-18
Applicant: AJINOMOTO CO., INC.
Inventor： Chihiro Tsuji , Yasuhiro Mihara , Shogo Nakano , Tomoharu Motoyama , Souhei Ito
Abstract: The present invention provides a method for producing a heparosan compound having an isomerized hexuronic acid residue and a method for producing a heparan sulfate with improved C5-epimerization efficiency. Specifically, the present invention provides a method for producing a heparosan compound having an isomerized hexuronic acid residue, said method comprising producing the heparosan compound having the isomerized hexuronic acid residue from a heparosan compound in the presence of a protein selected from the group of consisting of the following (A) to (F): (A) a protein comprising the amino acid sequence of SEQ ID No:2; (B) a protein which comprises an amino acid sequence having 90% or more homology to the amino acid sequence of SEQ ID No:2 and has a D-glucuronyl C5-epimerase substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID No:2 and has a D-glucuronyl C5-epimerase activity; (D) a protein comprising the amino acid sequence of SEQ ID NO:5; (E) a protein which comprises an amino acid sequence having 90% or more homology to the amino acid sequence of SEQ ID No:5 and has a D-glucuronyl C5-epimerase activity; and (F) a protein which comprises an amino acid sequence having one or several deleted, substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID No:5 and has a D-glucuronyl C5-epimerase activity.
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公开(公告)号：US11525132B2
公开(公告)日：2022-12-13
申请号：US17210094
申请日：2021-03-23
Applicant: AJINOMOTO CO., INC.
Inventor： Miwa Konishi , Shohei Kajimoto , Yusuke Hagiwara , Yasuhiro Mihara , Shogo Nakano , Tomoharu Motoyama , Sohei Ito
Abstract: Ligase mutants of the following (1), (2), or (3):
(1) a ligase mutant comprising an amino acid sequence showing 95% or more identity to the amino acid sequence of SEQ ID NO: 1, and having a nucleic acid-linking activity;
(2) a ligase mutant comprising an amino acid sequence showing 90% or more identity to the amino acid sequence of SEQ ID NO: 2, and having a nucleic acid-linking activity; or
(3) a ligase mutant comprising an amino acid sequence showing 97% or more identity to the amino acid sequence of SEQ ID NO: 3, and having a nucleic acid-linking activity, have excellent properties.-
4.发明申请公开(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
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.
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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
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.
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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
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.
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