Ajinomoto Co., Ltd.

Chihiro Tsuji

Yasuhiro Mihara

Shogo Nakano

Tomoharu Motoyama

Masayuki Sugiki

Uno Tagami

Tomoko Shimizu

Sohei Ito

Hiroyuki Nozaki

Souhei Ito

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

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.

Method for producing heparosan compound having isomerized hexuronic acid residue

&#x3b3;-Glu-Val synthetase suitable for generating &#x3b3;-Glu-Val, and a method for producing &#x3b3;-Glu-Val-Gly using the same are provided. By using &#x3b3;-Glu-Val synthetase showing a ratio of &#x3b3;-glutamylvaline synthetase activity to &#x3b3;-glutamylglycine synthetase activity of 2.0 or higher, such as &#x3b3;-Glu-Val synthetase of(AJ3132), &#x3b3;-Glu-Val-Gly is produced from Glu, Val, and Gly as raw materials.

Gamma glutamyl-valine synthase, and method for producing gamma glutamyl-valyl-glycine

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Kawasaki, Japan

Chihiro Tsuji