Enzymatic production of D-allulose
The current disclosure provides a process for enzymatically converting a saccharide into allulose. The invention also relates to a process for preparing allulose where the process involves converting fructose 6-phosphate (F6P) to allulose 6-phosphate (A6P), catalyzed by allulose 6-phosphate 3-epimerase (A6PE), and converting the A6P to allulose, catalyzed by allulose 6-phosphate phosphatase (A6PP).
1. An enzymatic process for preparing D-allulose from a starch derivative, the process comprising the steps of:
(i) converting the starch derivative to glucose 1-phosphate (G1P), catalyzed by an alpha glucan phosphorylase (αGP);
(ii) converting the G1P to glucose 6-phosphate (G6P), catalyzed by a phosphoglucomutase (PGM);
(iii) converting the G6P to fructose 6-phosphate (F6P), catalyzed by a phosphoglucoisomerase (PGI);
(iv) converting the F6P to allulose 6-phosphate (A6P), catalyzed by an allulose 6-phosphate epimerase (A6PE); and
(v) converting the A6P to D-allulose, catalyzed by an allulose 6-phosphate phosphatase (A6PP), wherein the A6PP dephosphorylate the A6P to produce the D-allulose and phosphate ions.
2. The process of claim 1 , wherein the starch derivative is prepared by:
enzymatic hydrolysis of starch, catalyzed by an isoamylase, a pullulanase, an alpha-amylase, or a combination thereof; or
acid hydrolysis of starch.
3. The process of claim 1 , wherein the process steps are conducted under at least one of the following process conditions:
at a temperature ranging
from 37° C. to 85° C.; and/or at a pH ranging from 5.0 to 8.0.
4. The process of claim 1 , wherein the process steps are conducted under at least one of the following process conditions:
without adenosine triphosphate (ATP) as a source of phosphate;
without nicotinamide adenosine dinucleotide (NAD); and
at a phosphate concentration from about 0.1 mM to about 150 mM,
wherein:
phosphate is recycled; wherein the step (v) involves an energetically favorable chemical reaction; and wherein the steps (i) to (v) are conducted in a single reaction vessel.
5. The process of claim 1 , wherein the phosphate ions produced by A6PP dephosphorylation of the A6P are used in the process step of converting the starch derivative to G1P.
6. The process of claim 1 , wherein the D-allulose is separated from reaction components and degradation products of the steps (i-v) without using chromatography.
7. The process of claim 1 , wherein the starch derivative is selected from the group consisting of amylose, amylopectin, soluble starch, amylodextrin, maltotriose, maltodextrin, maltose, and glucose.