US 11,639,513 B2Granted

Method for producing heparosan compound having isomerized hexuronic acid residue

AssigneeAjinomoto Co., Inc.

InventorsChihiro Tsuji, Yasuhiro Mihara, Shogo Nakano, Tomoharu Motoyama, Souhei Ito

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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.

A protein selected from the group consisting of:

(1) a protein comprising the amino acid sequence of SEQ ID NO. 56;
(2) a protein which comprises an amino acid sequence having 97% or more identity to the amino acid sequence of SEQ ID NO. 56 and has a D-glucuronyl C5-epimerase activity;
and
(3) a protein which comprises an amino acid sequence having 1 to 10 deleted, substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID NO. 56 and has a D-glucuronyl C5-epimerase activity;
(4) a protein comprising the amino acid sequence of SEQ ID 59;
(5) a protein which comprises an amino acid sequence having 97% or more identity to the amino acid sequence of SEQ ID 59 and has a D-glucuronyl C5-epimerase activity;
(6) a protein which comprises an amino acid sequence having I to 10 deleted, substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID 59 and has a D-glucuronyl C5-epimerase activity;
(7) a protein comprising the amino acid sequence of SEQ ID 62;
(8) a protein which comprises an amino acid sequence having 95% or more identity to the amino acid sequence of SEQ ID 62 and has a D-glucuronyl C5-epimerase activity;
(9) a protein which comprises an amino acid sequence having 1 to 25 deleted substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID 62 and has a D-glucuronyl C5-epimerase activity;
(10) a protein comprising the amino acid sequence of SEQ ID 65;
(11) a protein which comprises an amino acid sequence having 97% or more identity to the amino acid sequence of SEQ ID 65 and has a D-glucuronyl C5-epimerase activity;
(12) a protein which comprises an amino acid sequence having 1 to 10 deleted, substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID 65 and has a D-glucuronyl C5-epimerase activity;
(13) a protein comprising the amino acid sequence of SEQ ID 68;
(14) a protein which comprises an amino acid sequence having 99% or more identity to the amino acid sequence of SEQ ID 68 and has a D-glucuronyl C5-epimerase activity;
and
(15) a protein which comprises an amino acid sequence having 1 to 5 deleted, substituted, added or inserted amino acid residues in the amino acid sequence of SEQ ID 68 and has a D-glucuronvl C5-epimerase activity.

14 more elements

Facts

Application

16/515,415Filed 18 Jul 2019 · continuation

Priority

JP JP2017-00790019 Jan 2017

Published

US 2019/0338330 A17 Nov 2019

Granted

2 May 2023

Term ends

Not estimatedThe source records don’t support an estimate.

Classification9 codes

C08B 37/0075
Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan: heparin; Heparan sulfate; Derivatives…
C12P 19/26
Preparation of compounds containing saccharide radicals: preparation of nitrogen-containing carbohydrates
C12N 9/90
Enzymes; Proenzymes; Compositions thereof ; Processes for preparing, activating, inhibiting, separating or purifying enzymes: isomerases
C12N 15/09
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or…:…

Abstract

Claims14