Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes

Patent metadata

Field	Value
Publication number	US-9982284-B2
Application number	US-201514631931-A
Country	US
Kind code	B2
Filing date	Feb 26, 2015
Priority date	Feb 27, 2014
Publication date	May 29, 2018
Grant date	May 29, 2018

Abstract

A method is disclosed for hydrolyzing an alpha-1,5 glucosyl-fructose linkage in a saccharide (disaccharide or oligosaccharide) such as leucrose. This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transglucosidase or glucoamylase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one alpha-1,5 glucosyl-fructose linkage of the saccharide. This method is useful for reducing the amount of leucrose in a filtrate isolated from a glucan synthesis reaction, for example.

First claim

What is claimed is:

1. A method of reducing the amount of leucrose present in (i) a glucan synthesis reaction, or (ii) a fraction thereof, wherein said method comprises:
contacting said glucan synthesis reaction or said fraction thereof with a transglucosidase, wherein the amount of said leucrose is reduced in said glucan synthesis reaction or said fraction thereof compared to the amount of said leucrose that was present prior to said contacting, and wherein said leucrose is a byproduct of said glucan synthesis reaction.

2. The method of claim 1 , wherein said transglucosidase is immobilized.

3. The method of claim 1 , wherein the concentration of leucrose after said contacting is less than 50% of the concentration of leucrose that was present prior to said contacting.

4. The method of claim 1 , wherein said glucan synthesis reaction produces at least one insoluble alpha-glucan product.

5. The method of claim 1 , wherein said glucan synthesis reaction produces at least one soluble alpha-glucan product that is:
(i) a product of a glucosyltransferase, or
(ii) a product of the concerted action of both a glucosyltransferase and an alpha-glucanohydrolase capable of hydrolyzing glucan polymers having one or more alpha-1,3-glycosidic linkages or one or more alpha-1,6-glycosidic linkages.

6. The method of claim 1 , wherein said fraction is a chromatographic fraction of said glucan synthesis reaction.

7. A method of enriching fructose present in a fraction of a glucan synthesis reaction, comprising:
(a) producing a fraction of said glucan synthesis reaction according to the method of claim 1 , thereby providing a hydrolyzed fraction containing fructose; and
(b) separating fructose from said hydrolyzed fraction of step (a) to obtain a composition having a higher concentration of fructose compared to the fructose concentration of said fraction of step (a).

8. A fermentation method comprising:
(a) producing a fraction of said glucan synthesis reaction according to the method of claim 1 ;
(b) fermenting said fraction of step (a) with a microbe to yield a product, wherein said fermenting is performed after step (a) or simultaneously with step (a); and
(c) optionally, isolating said product of step (b);
wherein the yield of said product of (b) is increased compared to the product yield of fermenting a fraction of said glucan synthesis reaction that has not been contacted with said transglucosidase.

9. The method of claim 4 , wherein at least 50% of the glycosidic linkages of said insoluble alpha-glucan product are alpha-1,3-glycosidic linkages.

10. The method of claim 9 , wherein at least 95% of the glycosidic linkages of said insoluble alpha-glucan product are alpha-1,3-glycosidic linkages.

11. The method of claim 9 , wherein said insoluble alpha-glucan product has a number average degree of polymerization of at least 100.

12. The method of claim 4 , wherein said method comprises contacting said glucan synthesis reaction with said transglucosidase.

13. The method of claim 4 , wherein said method comprises contacting said fraction with said transglucosidase.

14. The method of claim 13 , wherein said fraction is a filtrate of said glucan synthesis reaction.

15. The method of claim 13 , wherein said fraction is a supernatant of the glucan synthesis reaction.

16. The method of claim 1 , wherein said method comprises contacting said glucan synthesis reaction with said transglucosidase.

17. The method of claim 9 , wherein at least 80% of the glycosidic linkages of said insoluble alpha-glucan product are alpha-1,3-glycosidic linkages.

18. The method of claim 1 , wherein said glucan synthesis reaction is an isolated glucan synthesis reaction.

19. The method of claim 1 , wherein said method comprises contacting said fraction with said transglucosidase.

Assignees

• Du Pont

Inventors

• Nagy Kevin D
• Hago Erwin Columbus
• Shetty Jayarama K
• Hennessey Susan Marie
• Dicosimo Robert
• Hua Ling
• Ramirez Rodrigo
• Tang Zhongmei
• Yu Zheyong

Classifications

• C12P19/16

  produced by the action of an alpha-1, 6-glucosidase, e.g. amylose, debranched amylopectin (non-biological hydrolysis of starch C08B30/00) · CPC title

• C12P19/04

  Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds · CPC title

• C12P19/18

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

• C12P19/02

  Monosaccharides (2-ketogulonic acid C12P7/60) · CPC title

• C12P19/14 Primary

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title