Glucosyl Stevia composition
Glucosyl stevia compositions are prepared from steviol glycosides of Stevia rebaudiana Bertoni. The glucosylation was performed by cyclodextrin glucanotransferase using the starch as source of glucose residues. The short-chain glucosyl stevia compositions were purified to >95% content of total steviol glycosides. The compositions can be used as sweetness enhancers, flavor enhancers and sweeteners in foods, beverages, cosmetics and pharmaceuticals.
1. A sweetener composition comprising a highly purified glucosyl stevia composition made by the process of:
(a) adding starch into water to form a starch suspension;
(b) adding a mixture of α-amylase and CGTase into the starch suspension and incubating for about 0.5 to 2 hours at about 75-80° C., resulting in a liquefied starch suspension;
(c) inactivating the α-amylase by low pH heat treatment; and
cooling the liquefied starch suspension and adjusting the pH to about 5.5 to 7.0;
(d) adding steviol glycosides into the liquefied starch suspension, resulting in a reaction mixture;
(e) adding a second batch of CGTase into the reaction mixture and incubating for about 12 to 48 hours at about 55-75° C.;
(f) adding β-amylase to reaction mixture and incubating for about 12-24 hours at about 35-55° C.;
(g) inactivating the enzymes in the reaction mixture by heat treatment;
(h) decolorizing the reaction mixture;
(i) removing non-diterpene compounds by contacting the decolorized reaction mixture with macroporous adsorbent resin and subsequently eluting adsorbed diterpene glycosides with aqueous ethanol to result in a glycoside-containing aqueous ethanol eluate;
(j) desalting the glycoside-containing aqueous ethanol eluate with ion-exchange resins;
(k) removing ethanol from the aqueous ethanol eluate, resulting in an aqueous eluate;
(l) concentrating and drying the aqueous eluate to obtain the dried glucosyl stevia composition,
(m) suspending the dried glucosyl stevia composition in aqueous alcohol to form a suspension, removing unmodified steviol glycosides from the suspension, and separating and drying the highly purified glucosyl stevia composition;
wherein the highly purified glucosyl stevia composition comprises greater than about 50% by weight mono- and diglucosyl steviol glycosides, and less than about 15% unmodified steviol glycosides, and wherein the sweetener composition comprises an additional sweetening agent selected from the group consisting of: stevia extract, steviol glycosides, stevioside, Rebaudioside A, Rebaudioside B, Rebaudioside C, Rebaudioside D, Rebaudioside E, Rebaudioside F, dulcoside A, steviolbioside, rubusoside, other steviol glycosides found in Stevia rebaudiana Bertoni plant and mixtures thereof, Luo Han Guo extract, mogrosides, high-fructose corn syrup, corn syrup, invert sugar, fructooligosaccharides, inulin, inulooligosaccharides, coupling sugar, maltooligosaccharides, maltodextins, corn syrup solids, glucose, maltose, sucrose, lactose, aspartame, saccharin, sucralose, sugar alcohols, and a combination thereof.
2. A flavor composition comprising a highly purified glucosyl stevia composition made by the process of:
(a) adding starch into water to form a starch suspension;
(b) adding a mixture of α-amylase and CGTase into the starch suspension and incubating for about 0.5 to 2 hours at about 75-80° C., resulting in a liquefied starch suspension;
(c) inactivating the α-amylase by low pH heat treatment; and
cooling the liquefied starch suspension and adjusting the pH to about 5.5 to 7.0;
(d) adding steviol glycosides into the liquefied starch suspension, resulting in a reaction mixture;
(e) adding a second batch of CGTase into the reaction mixture and incubating for about 12 to 48 hours at about 55-75° C.;
(f) adding β-amylase to reaction mixture and incubating for about 12-24 hours at about 35-55° C.;
(g) inactivating the enzymes in the reaction mixture by heat treatment;
(h) decolorizing the reaction mixture;
(i) removing non-diterpene compounds by contacting the decolorized reaction mixture with macroporous adsorbent resin and subsequently eluting adsorbed diterpene glycosides with aqueous ethanol to result in a glycoside-containing aqueous ethanol eluate;
(j) desalting the glycoside-containing aqueous ethanol eluate with ion-exchange resins;
(k) removing ethanol from the aqueous ethanol eluate, resulting in an aqueous eluate;
(l) concentrating and drying the aqueous eluate to obtain the dried glucosyl stevia composition,
(m) suspending the dried glucosyl stevia composition in aqueous alcohol to form a suspension, removing unmodified steviol glycosides from the suspension, and separating and drying the highly purified glucosyl stevia composition;
wherein the highly purified glucosyl stevia composition comprises greater than about 50% by weight mono- and diglucosyl steviol glycosides, and less than about 15% unmodified steviol glycosides, and wherein the sweetner composition comprises an additional flavoring agent selected from the group consisting of: lemon, orange, fruity, banana, grape, pear, pineapple, mango, bitter almond, cola, cinnamon, sugar, cotton candy, vanilla, and a combination thereof.
3. A food ingredient comprising a highly purified glucosyl stevia composition made by the process of:
(a) adding starch into water to form a starch suspension;
(b) adding a mixture of α-amylase and CGTase into the starch suspension and incubating for about 0.5 to 2 hours at about 75-80° C., resulting in a liquefied starch suspension;
(c) inactivating the α-amylase by low pH heat treatment; and
cooling the liquefied starch suspension and adjusting the pH to about 5.5 to 7.0;
(d) adding steviol glycosides into the liquefied starch suspension, resulting in a reaction mixture;
(e) adding a second batch of CGTase into the reaction mixture and incubating for about 12 to 48 hours at about 55-75° C.;
(f) adding β-amylase to reaction mixture and incubating for about 12-24 hours at about 35-55° C.;
(g) inactivating the enzymes in the reaction mixture by heat treatment;
(h) decolorizing the reaction mixture;
(i) removing non-diterpene compounds by contacting the decolorized reaction mixture with macroporous adsorbent resin and subsequently eluting adsorbed diterpene glycosides with aqueous ethanol to result in a glycoside-containing aqueous ethanol eluate;
(j) desalting the glycoside-containing aqueous ethanol eluate with ion-exchange resins;
(k) removing ethanol from the aqueous ethanol eluate, resulting in an aqueous eluate;
(l) concentrating and drying the aqueous eluate to obtain the dried glucosyl stevia composition,
(m) suspending the dried glucosyl stevia composition in aqueous alcohol to form a suspension, removing unmodified steviol glycosides from the suspension, and separating and drying the highly purified glucosyl stevia composition;
wherein the highly purified glucosyl stevia composition comprises greater than about 50% by weight mono- and diglucosyl steviol glycosides, and less than about 15% unmodified steviol glycosides, and wherein the sweetner composition comprises an additional food ingredient selected from the group consisting of: acidulants, organic and amino acids, coloring agents, bulking agents, modified starches, gums, texturizers, preservatives, antioxidants, emulsifiers, stabilisers, thickeners, gelling agents, and a combination thereof.
4. A food, beverage, cosmetic or pharmaceutical product comprising a highly purified glucosyl stevia composition made by the process of:
(a) adding starch into water to form a starch suspension;
(b) adding a mixture of α-amylase and CGTase into the starch suspension and incubating for about 0.5 to 2 hours at about 75-80° C., resulting in a liquefied starch suspension;
(c) inactivating the α-amylase by low pH heat treatment; and
cooling the liquefied starch suspension and adjusting the pH to about 5.5 to 7.0;
(d) adding steviol glycosides into the liquefied starch suspension, resulting in a reaction mixture;
(e) adding a second batch of CGTase into the reaction mixture and incubating for about 12 to 48 hours at about 55-75° C.;
(f) adding β-amylase to reaction mixture and incubating for about 12-24 hours at about 35-55° C.;
(g) inactivating the enzymes in the reaction mixture by heat treatment;
(h) decolorizing the reaction mixture;
(i) removing non-diterpene compounds by contacting the decolorized reaction mixture with macroporous adsorbent resin and subsequently eluting adsorbed diterpene glycosides with aqueous ethanol to result in a glycoside-containing aqueous ethanol eluate;
(j) desalting the glycoside-containing aqueous ethanol eluate with ion-exchange resins;
(k) removing ethanol from the aqueous ethanol eluate, resulting in an aqueous eluate;
(l) concentrating and drying the aqueous eluate to obtain the dried glucosyl stevia composition,
(m) suspending the dried glucosyl stevia composition in aqueous alcohol to form a suspension, removing unmodified steviol glycosides from the suspension, and separating and drying the highly purified glucosyl stevia composition;
wherein the highly purified glucosyl stevia composition comprises greater than about 50% by weight mono- and diglucosyl steviol glycosides, and less than about 15% unmodified steviol glycosides.