Inositol preparation method
An inositol preparation method by enzymatic catalysis uses starch and cellulose or substrates thereof as substrates. Raw materials are converted to inositol by in vitro multi-enzyme reaction system in one pot. The yield from the substrate to inositol is significantly improved by process optimization and adding new enzymes. The new enzymes can promote the phosphorolysis of starch or cellulose and utilization of glucose, which is the final production after the phosphorolysis of starch and cellulose. The inositol preparation method described herein has great potentials in industrial production of inositol because of high inositol yield, easy scale-up, low production cost, and lower impact to environment.
1. A method for preparing inositol, comprising:
(1) using starch or starch derivative as substrate, adding α-glucan phosphorylase, phosphoglucomutase, Archaeoglobus fulgidus inositol 3-phosphate synthase and Thermatoga maritime inositol monophosphatase to establish an in vitro multi-enzyme reaction system and perform an enzyme-catalyzed reaction; and
(2) separating and purifying reaction product to obtain inositol, wherein the starch derivative of step (1) is selected from the group consisting of partially hydrolyzed starch, starch dextrin, maltodextrin, malto-oligosaccharide, and maltose,
wherein the enzyme-catalyzed reaction is performed at 60-80° C., and is performed in one pot,
wherein the multi-enzyme reaction system further comprises starch debranching enzyme, amylase, and one or both of maltose phosphorylase and glucanotransferase,
wherein the starch debranching enzyme is one or both of isoamylase and pullulanase,
wherein the multi-enzyme reaction system further comprises polyphosphate glucokinase and polyphosphate, wherein the polyphosphate is sodium polyphosphate.
2. The preparation method according to claim 1 , wherein the multi enzyme reaction system further comprises the following components: buffer, inorganic phosphate anion, divalent magnesium ion, and either zinc ion or manganese ion.
3. A method for preparing inositol from soluble starch, comprising:
(1) treating the soluble starch with pullulanase at 37° C. to obtain a starch derivative;
(2) using the starch derivative as a substrate, adding α-glucan phosphorylase, phosphoglucomutase, Archaeoglobus fulgidus inositol 3-phosphate synthase and Thermatoga maritime inositol monophosphatase to establish an in vitro multi-enzyme reaction system and perform an enzyme-catalyzed reaction; and
(3) separating and purifying reaction product to obtain inositol,
wherein the enzyme-catalyzed reaction is performed at 60-80° C., and is performed in one pot,
wherein the multi-enzyme reaction system further comprises isoamylase, amylase, and one or both of maltose phosphorylase and glucanotransferase,
wherein the multi-enzyme reaction system further comprises polyphosphate glucokinase and polyphosphate;
wherein the polyphosphate is sodium polyphosphate.