	US 11,919,919 B2
	Synthesis of oligosaccharides
Nikolay Khanzhin, Humlebæk (DK); and Markus Jondelius Hederos, Trelleborg (SE)
Assigned to GLYCOM A/S, Hørsholm (DK)
Filed by GLYCOM A/S, Hørsholm (DK)
Filed on Dec. 27, 2021, as Appl. No. 17/646,093.
Application 17/646,093 is a continuation of application No. 16/626,245, granted, now 11,214,588, previously published as PCT/IB2018/054753, filed on Jun. 27, 2018.
Claims priority of application No. 2017 70522 (DK), filed on Jun. 30, 2017; application No. 2017 70523 (DK), filed on Jun. 30, 2017; and application No. 2017 70524 (DK), filed on Jun. 30, 2017.
Prior Publication US 2022/0119437 A1, Apr. 21, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C07H 1/06 (2006.01); A23L 3/00 (2006.01); A23L 33/00 (2016.01); B01D 61/02 (2006.01); B01D 61/04 (2006.01); B01D 61/14 (2006.01); B01D 61/58 (2006.01); B01D 69/02 (2006.01); B01D 69/12 (2006.01); B01D 71/56 (2006.01); B01D 71/58 (2006.01); C07H 3/06 (2006.01); C08B 37/00 (2006.01); C12P 19/04 (2006.01); C12P 19/12 (2006.01); C12P 19/18 (2006.01)

CPC C07H 1/06 (2013.01) [A23L 33/40 (2016.08); B01D 61/027 (2013.01); B01D 61/04 (2013.01); B01D 61/146 (2022.08); B01D 61/58 (2013.01); B01D 69/02 (2013.01); B01D 69/12 (2013.01); B01D 71/56 (2013.01); B01D 71/58 (2013.01); C07H 3/06 (2013.01); C08B 37/0003 (2013.01); C12P 19/04 (2013.01); C12P 19/12 (2013.01); C12P 19/18 (2013.01); A23L 33/00 (2016.08); A23V 2002/00 (2013.01); B01D 61/145 (2013.01); B01D 2315/16 (2013.01); B01D 2325/20 (2013.01)]

20 Claims

1. A method for producing a compound Gly-A, comprising the steps of:

a) initiating an enzymatic reaction of compound A and glycosylated compound B (Gly-B) in the presence of a glycosidase capable of transferring the glycosyl (Gly) moiety to compound A;

b) allowing the reaction in step a) to reach equilibrium to form a first mixture comprising compound A, Gly-B, glycosylated compound A (Gly-A), and compound B;

c) contacting the mixture with a nanofiltration membrane with a molecular weight cut-off (MWCO) of 600-3500 Da allowing at least a part of compound B to pass and ensuring the retention of compounds Gly-A, compound A and Gly-B, wherein the active (top) layer of the membrane is composed of polyamide, and wherein the MgSO4 rejection of the membrane is 50-90%;

d) re-initiating an enzymatic reaction of compound A and Gly-B of the retentate the presence of Gly-A and a glycosidase capable of transferring the glycosyl (Gly) moiety to compound A to form a second mixture;

e) collecting the Gly-A from the second mixture;

wherein compound A is a tri- or higher oligosaccharide, Gly-A is compound A glycosylated with the Gly moiety, compound B is a disaccharide, and GlyB is compound B glycosylated with the Gly moiety, provided that compounds A and Gly-B are not identical.