Method for converting sucrose to β-D-glucose

• US 7,556,945 B1
• Filed: 07/05/2007
• Issued: 07/07/2009
• Est. Priority Date: 03/14/2005
• Status: Active Grant

• Current Assignee: National Technology & Engineering Solutions Of Sandia LLC (Honeywell International Inc.)
• Original Assignee: Sandia Corporation (Martin Marietta Corporation)
• Inventors: Walker, Andrew, Volponi, Joanne V., Ingersoll, David, Simmons, Blake A.
• Primary Examiner(s): Lilling; Herbert J.
• Application Number:
  US11/825,615
• Time in Patent Office: 733 Days
• Field of Search: None
• US Class Current: 435/94
• CPC Class Codes: C12P 19/02 Monosaccharides 2-ketogulonic acid C12P7/60
• CPC Class Codes: C12P 41/001 by metabolizing one of the enantiomers

Abstract

Disclosed is an apparatus and method for continuously converting sucrose to β-D-glucose. The method comprises a three-stage enzymatic reactor in which an aqueous solution of sucrose is first converted into a solution of fructose and α-D-glucose by passing it through a porous, packed column containing an inert media on which invertase is immobilized. This solution is then sent through a second packed column containing glucose isomerase and finally a third packed column containing mutarotase. Solution temperature and pH are adjusted to maximize glucose output.

Claims

1. A method for converting sucrose to β

-D-glucose, comprising the steps of;

a) providing a sucrose containing solution having a pH of between about 5.8 and about 4.6;

b) heating the sucrose containing solution to a temperature of about 43°

C. to about 47°

C.;

c) contacting the sucrose containing solution with invertase, wherein the conditions of pH and temperature are maintained, to provide a fructose and α

-D-glucose containing solution;

d) contacting the fructose and α

-D-glucose containing solution with glucose isomerase, wherein the conditions of pH and temperature are maintained, to provide a solution containing α

-D-glucose and β

-D-glucose; and

e) contacting the α

-D-glucose and β

-D-glucose with mutarotase, wherein the conditions of pH and temperature are maintained, to provide a solution containing a high percentage of β

-D-glucose.

2. The method of claim 1, wherein the step of contacting the fructose and α

-D-glucose containing solution with glucose isomerase, further comprises stabilizing the glucose isomerase with a Mg⁺ stabilizer.

3. The method of claim 2, wherein the Mg⁺ stabilizer comprises MgCl₂.

4. The method of claim 3, wherein MgCl₂ is present in a concentration of about 20 mM.

5. The method of claim 1, wherein said invertase, said glucose isomerase, and said mutarotase are each contained on separate solid substrates having a plurality of aminated structures attached thereto.

6. The method of claim 5, wherein the separate solid substrates comprise a plurality of silica beads.

7. The method of claim 6, wherein each of the plurality of aminated structures comprises an amine functional group attached to said native surface of the silica beads, and an amine group cross-linker attached to each of said amine functional groups, wherein said amine group cross-linker provides a site for protein reaction.

8. The method of claim 7, wherein the amine functional group is selected from the list consisting of aminopropyltriethoxysilane, aminopropyltrimethoxysilane, aminopropylmethyldiethoxysilane;

aminopropylmethyldimethoxysilane;

aminoethylaminopropyltriethoxysilane;

aminoethylaminopropyltrimethoxysilane;

aminoethylaminopropylmethyldimethoxysilane;

diethylenetriaminopropyltriethoxysilane;

diethylenetriaminopropyltrimethoxysilane;

diethylenetriaminopropylmethyldiethoxysilane;

diethylenetriaminopropylmethyldimethoxysilane;

cyclohexylaminopropyltrimethoxysilane;

hexanediaminomethyltriethoxysilane;

phenylaminomethyltriethoxysilane;

phenylaminomethyltrimethoxysilane;

diethylaminomethyltriethoxysilane;

(dimethylaminomethyl)methyldimethoxysilane; and

methylaminopropylltimethoxysilane.

9. The method of claim 7, wherein the plurality of amine functional groups comprise an amino alkyl alkyloxy silane.

10. The method of claim 9, wherein the plurality of amine functional groups comprise 3-aminopropyltriethoxysilane.

11. The method of claim 7, wherein the amine group cross-linker is selected from the group consisting of formaldehyde, glutaraldehyde, mono- and di-aldehyde PEG, and bis(imido esters), bis(succinimidyl esters), diisocyanates and diacid chlorides.

12. The method of claim 11, wherein the amine group cross-linker comprises a mono- or di-aldehyde.

13. The method of claim 12, wherein the amine group cross-linker is glutaraldehyde.