Preparation method for various novel fucosyl oligosaccharides and use thereof

Patent metadata

Field	Value
Publication number	US-11504386-B2
Application number	US-201816765359-A
Country	US
Kind code	B2
Filing date	Nov 15, 2018
Priority date	Nov 20, 2017
Publication date	Nov 22, 2022
Grant date	Nov 22, 2022

Abstract

The present invention relates to a preparation method for various novel fucosyl oligosaccharides and a use thereof. More specifically, the present invention allows a preparation of various novel fucosyl oligosaccharides through an enzymatic reaction with α-1,2-fucosyltransferase using a GDP-L-fucose donor and various glucose acceptors and an establishment of probiotic characteristics thereof, and thus has an effect of providing uses as materials for medicines, food, cosmetic products, and the like.

First claim

What is claimed is:

1. A method of producing a fucosyl-oligosaccharide comprising
reacting a sugar acceptor and a guanosine 5′-diphospho-p-L-fucose (GDP-L-fucose) donor with α-1,2-fucosyltransferase to produce a fucosyl-oligosaccharide,
wherein the sugar acceptor is selected from the group consisting of glucose, galactose, cellobiose, lactose, fructose, sucrose, maltose, mannose, xylose, and agarobiose,
the α-1,2-fucosyltransferase is selected from a sequence comprising the amino acid sequence of SEQ ID NO: 2, and
the fucosyl-oligosaccharide is selected from the group consisting of 2′-fucosylglucose, 2′-fucosylgalactose, 2′-fucosylcellobiose, 2′-fucosylfructose, 2′-fucosylsucrose, 2′-fucosylmaltose, 2′-fucosylmannose, 2′-fucosylxylose, and 2′-fucosylagarobiose, wherein the reacting is performed in the presence of a vector for expressing a phosphomannomutase (ManB), a mannose 1-phosphate guanylytransferase (ManC), a GDP-D-mannose-4,6-dehydratse (Gmd) and a GDP-4-keto-6-deoxymannose 3,5-epimerase 4-reductase (WcaG) for producing the GDP-L-fucose.

2. The method of claim 1 , wherein the reacting is performed at 20° C. to 40° C. for 3 hours to 24 hours.

3. A method of producing a fucosyl-oligosaccharide comprising
culturing, in the presence of a sugar acceptor and glycerol, recombinant E. coli or yeast into which a vector for expressing ManB, ManC, Gmd, and WcaG involved in the de novo pathway for producing a guanosine 5′-diphospho-β-L-fucose (GDP-L-fucose) donor, and a vector for expressing α-1,2-fucosyltransferase are introduced, and
separating and purifying the fucosyl-oligosaccharide from the E. coli or yeast culture,
wherein the sugar acceptor is selected from the group consisting of glucose, galactose, cellobiose, lactose, fructose, sucrose, maltose, mannose, xylose, and agarobiose, and
the α-1,2-fucosyltransferase is selected from a sequence comprising the amino acid sequence of SEQ ID NO: 2.

4. The method of claim 3 , wherein the culturing is fed-batch culture.

5. The method of claim 4 , wherein the fed-batch culture is performed at 25° C. to 37° C. for 2 hours to 60 hours, and isopropyl β-D-1-thiogalactopyranoside (IPTG) is added thereto, followed by further fed-batch culture at 20° C. to 30° C. for 60 hours to 150 hours.

Assignees

• Univ Korea Res & Bus Found

Inventors

• Kim Kyoung Heon
• Jin Yong-Su
• Yun Eun-Ju
• Yu Sora
• Lee Jaewon

Classifications

• A61Q19/008

  Preparations for oily skin · CPC title

• C12P19/18

  produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins · CPC title

• C12N9/1051

  Hexosyltransferases (2.4.1) · CPC title

• A23L33/125

  containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates (indigestible substances A23L33/21) · CPC title

• A61K31/729

  Agar; Agarose; Agaropectin · CPC title