Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes

Patent metadata

Field	Value
Publication number	US-10428362-B2
Application number	US-201815976246-A
Country	US
Kind code	B2
Filing date	May 10, 2018
Priority date	Feb 27, 2014
Publication date	Oct 1, 2019
Grant date	Oct 1, 2019

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 a saccharide present in (i) a glucan synthesis reaction, or (ii) a fraction thereof, wherein said method comprises:
contacting said glucan synthesis reaction or fraction thereof with an alpha-glucosidase enzyme, wherein the amount of said saccharide is reduced in said glucan synthesis reaction or said fraction thereof compared to the amount of said saccharide that was present prior to said contacting,
wherein said glucan synthesis reaction produces an insoluble alpha-glucan product, wherein at least 97% of the glycosidic linkages of said insoluble alpha-glucan product are alpha-1,3-glycosidic linkages, and
wherein said saccharide is a byproduct of said glucan synthesis reaction and is (a) leucrose or (b) a disaccharide or oligosaccharide that comprises at least one alpha-1,3 or alpha-1,6 glucosyl-glucose linkage.

2. The method of claim 1 , wherein said alpha-glucosidase enzyme is immobilized.

3. The method of claim 1 , wherein said saccharide is leucrose.

4. The method of claim 1 , wherein said saccharide comprises at least one alpha-1,3 or alpha-1,6 glucosyl-glucose linkage.

5. The method of claim 4 , wherein said saccharide comprises at least one alpha-1,3 glucosyl-glucose linkage.

6. The method of claim 1 , wherein said alpha-glucosidase enzyme is a transglucosidase or glucoamylase.

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 alpha-glucosidase enzyme.

9. The method of claim 1 , wherein at least 98% of glycosidic linkages of said insoluble alpha-glucan product are alpha-1,3-glycosidic linkages.

10. The method of claim 9 , wherein at least 99% of glycosidic linkages of said insoluble alpha-glucan product are alpha-1,3-glycosidic linkages.

11. The method of claim 10 , wherein at least 100% of glycosidic linkages of said insoluble alpha-glucan product are alpha-1,3-glycosidic linkages.

12. The method of claim 1 , wherein said insoluble alpha-glucan product has a number average degree of polymerization of at least 100.

13. The method of claim 1 , wherein said method comprises contacting said glucan synthesis reaction with said alpha-glucosidase enzyme.

14. The method of claim 1 , wherein said method comprises contacting said fraction with said alpha-glucosidase enzyme.

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

16. The method of claim 14 , wherein said fraction is a supernatant of the glucan synthesis reaction.

17. The method of claim 1 , wherein said alpha-glucosidase enzyme is a Neosartorya fischeri alpha-glucosidase enzyme, Rasamsonia composticola alpha-glucosidase enzyme, Bifidobacterium longum alpha-glucosidase enzyme, Bifidobacterium pseudolongum alpha-glucosidase enzyme, Bifidobacterium thermophilum alpha glucosidase enzyme, or Bifidobacterium breve alpha-glucosidase enzyme.

Assignees

• Du Pont

Inventors

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

Classifications

• C12P19/18

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

• C12P19/04

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

• C12P7/14

  Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms · CPC title

• C12P7/06

  Ethanol, i.e. non-beverage · 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