US 11,851,688 B2
2-O-sulfation enzyme mutant and 3-O-sulfation enzyme mutant, and method for using same
Chihiro Tsuji, Kanagawa (JP); Tomoko Shimizu, Kanagawa (JP); Uno Tagami, Kanagawa (JP); Yasuhiro Mihara, Kanagawa (JP); Masayuki Sugiki, Kanagawa (JP); Shogo Nakano, Shizuoka (JP); Tomoharu Motoyama, Shizuoka (JP); and Sohei Ito, Shizuoka (JP)
Assigned to AJINOMOTO CO., INC., Tokyo (JP)
Filed by AJINOMOTO CO., INC., Tokyo (JP)
Filed on Feb. 20, 2020, as Appl. No. 16/795,991.
Application 16/795,991 is a continuation of application No. PCT/JP2018/033897, filed on Sep. 5, 2018.
Claims priority of application No. 2017-170637 (JP), filed on Sep. 5, 2017.
Prior Publication US 2020/0181588 A1, Jun. 11, 2020
Int. Cl. C12N 9/10 (2006.01); C08B 37/00 (2006.01); C12P 19/26 (2006.01)
CPC C12N 9/13 (2013.01) [C08B 37/0075 (2013.01); C12P 19/26 (2013.01); C12Y 208/02 (2013.01)] 10 Claims
 
1. A 2-O-sulfation enzyme mutant comprising:
I) an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO: 2;
(b) an amino acid sequence comprising one to thirty 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 to twenty 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; and
(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;
II) a substitution of a leucine residue at position 321, relative to the amino acid sequence of SEQ ID NO: 2, with a basic amino acid residue; and
III) a 2-O-sulfate transfer activity.