CA1141226A - Low cariogenic food products - Google Patents
Low cariogenic food productsInfo
- Publication number
- CA1141226A CA1141226A CA000347168A CA347168A CA1141226A CA 1141226 A CA1141226 A CA 1141226A CA 000347168 A CA000347168 A CA 000347168A CA 347168 A CA347168 A CA 347168A CA 1141226 A CA1141226 A CA 1141226A
- Authority
- CA
- Canada
- Prior art keywords
- set forth
- food products
- sucrose
- aldosylfructoside
- foods
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/30—Artificial sweetening agents
- A23L27/33—Artificial sweetening agents containing sugars or derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
- C12N9/1051—Hexosyltransferases (2.4.1)
- C12N9/1055—Levansucrase (2.4.1.10)
Abstract
ABSTRACT OF THE DISCLOSURE
The present invention relates to a process for producing foods and drinks sweetened with sugar sweeteners.
More particularly, the present process is based on the invention that low-cariogenic and/or anti-caries foods and drinks are obtainable by using an aldosylfructoside obtained by subjecting a substrate solution containing aldose, and sucrose and/or raffinose to the action of levansucrase (E.C. 2.4.1.10).
Also, the process is based on the promising finding that said aldosylfructoside is usable in the production of all products which are used orally as well as foods and drinks in general.
The present invention relates to a process for producing foods and drinks sweetened with sugar sweeteners.
More particularly, the present process is based on the invention that low-cariogenic and/or anti-caries foods and drinks are obtainable by using an aldosylfructoside obtained by subjecting a substrate solution containing aldose, and sucrose and/or raffinose to the action of levansucrase (E.C. 2.4.1.10).
Also, the process is based on the promising finding that said aldosylfructoside is usable in the production of all products which are used orally as well as foods and drinks in general.
Description
The present invention relates to a process for producing foods and drinks.
Sucrose, a typical sweetener with sufficient sweetness and body, is consumed in large amount in a production of foods and drinks. Recentlv, it has been demonstrated that sweetened foods and drinks, particularly those sweetened with sucrose, cause very often dental caries. Dental caries is generally caused as follows; sucrose intaken is converted by oral bacteria into water-insoluble glucans such as dextran which adheres on tooth surface in layers, sugar intaken pass throu~h the layers and reach to the tooth surface where they are fermented anaerobically into organic acids, and the acids act on the enamel. Since sucrose is a major factor in causing dental caries, the realization of a process for producing foods and drinks using low-cariogenic sugars has been in great expectation.
According to the invention there is provided a process for producing food products, characterized in that the food products are prepared with, or added with aldosylfructoside which is obtained hy subjecting a 5 - 50 w/w% substrate solution, containing sucrose, raffinose or a mixture thereof, and one or more members selected from the group consisting of D-xylose, L-arabinose, D-galactose, D-mannose, xylobiose, cellobiose, maltose, isomaltosc, lactose, kojibiose, laminaribiose, nigerose, xylotriose, cellotriose, maltotriose, isomaltotriose, panose, isopanose, and partial hydrolysates of starch, dextran, pullulan, curdlan, pachyman, elsinan, glucomannan, cellulose and xylan, in the weight ratio of 1 : 50 - 5~ : 1, to the enzymatic action of levansucrase (E.C. 2.4.1.10) at a pH in the range _A_ cb/lJ , of 3 - 10 and a temperature in the range of 20 - 80C for 0.1 - 100 hours.
The present inventors investigated processes for producing low-cariogenic foods and drinks. The present invention is based on the discovery resulting from the efforts that aldosylfructoside obtalned by subjecting a substrate solution containing aldose, and sucrose and/or raffinose to the action of levansucrase (E.C. 2.4.1.10) has an appropriate sweetness and also low-cariogenic - la -cb/
ZG
and/or anti-caries properties which offer aldosylfructoside as a favorable sweetener for producing sweetened low-cariogenic foods and drinks.
Aldoses usable in the present invention are those which can be converted into aldosylfructosides excluding sucrose and raffinose by transfructosylation with levansuc~ase. Namely, aldosyl mono-, di- and tri-saccharides excluding glucose and melibiose are preferable: for example one or more members of a group comprising D-xylose, L-arabinose, D-galactose, D-mannose, xylobiose, cellobiose, maltose, isomaltose, lactose, kojibiose, laminaribiose, nigerose, xylotriose, cellotriose, maltotriose, isomaltotriose, panose and isopanose.
In addition, sugar mixtures, D.E. about 10 to 70, prepared by partial hydrolysis of polysaccharides such as starch, dextran, pullulan, curdlan9 pachyman, elsinan, glucomannan, cellulose and xylan, are usable in the present invention.
Substrate solution referred in the present invention is an aqueous solution containing aldose which acts as an acceptor for fructose residue, and sucrose and/or raffinose which act as a donor of fructose residue, upon transfructosylation reaction with levansucrase.
The mole ratio of acceptor vs. donor in the range of about 1 : 50 to 50 : 1, and substrate concentration in the range of about 5 to 50 w/w %, are preferable.
Levansucrase (E.C. 2.4.1.10) employable in the present invention is that which forms non-reducing aldosylfructosides when allowed to act on the above-described substrate solution containing aldose, and sucrose and/or raffinose, to transfer the fructosyl residues of sucrose and/or raffinose to the reducing 1141~Z6 Cl-sites of the aldoses. For example, levansucrase from Actinomyces v~scosus, Ae~obacter ~ev~icum, Acetobacter svboxydens, Boc~us Zichenifo~mis, Bac~us subti~is, G~uconobacter oxydens~ Streptococcus mutons, Streptococcus sa~ivaZius and other microorganisms is advantageously usable in the present invention.
In order to prepare levansucrase from the microorganisms, submerged culture is usually employed. The culture broth can be used without pre-treatment, but its supernatant or filtrate is usually used after removing water-insoluble substances by centrifugation or filtration. In some cases, the microbes can be used intact as an enzyme preparation without extraction, as well as levansucrase extracted from the cells by conventional methods. The levansucrase thus obtained is, if desired, purified further according to conventional method. In addition, immobilized levansucrases are also usable in continuous or batch-wise operation.
In the production of aldosylfructoside, a substrate solution containing aldose, and sucrose and/or raffinose is added with levansucrase to effect the enzymatic transfructosylation react;on. In this case, any reaction temperature and pH can be applied so far as the levansucrase acts on the substrate to produce corresponding aldosylfructoside; generally, the pH range of 3 to 10 and the temperature range of about 20 to 80CC are preferable. Levansucrase is used in the range of about 0.01 to 1,000 units per 9 sucrose and/or raffinose as defined by the following assay method, and the reaction is performed usually for about 0.1 to 100 hours.
Assay of levansucrase activitY
Two ml of a reaction mixture containing 10 w/v % sucrose, 50 mM
phosphate buffer, pH 7.0, and levansucrase is incubated at 30C for 30 minutes. The reaction mixture is then heated to inactivate the enzyme and the amount of the glucose released is determined by the glucose-oxidase method. One unit of levansucrase activity is defined as the amount of enzyme that produce 1 ~ mole of glucose per minute under the above conditions.
The reaction mixture containing aldosylfructoside can be, if desired, treated further with other enzymes such as glucose isomerase which isomerizes the glucose released in the reaction mixture into fructose and increases the sweetness of the product.
The transfructosylation reaction is usually suspended by heating and the reaction mixture i 5 filtrated. The filtrate is decolorized with activated carbon and deionized with ion exchangers, for example those of H-form and OH-form. The purified solution containing aldosylfructoside is concentrated into syrup, or dried and pulverized into powder for use in the production of foods and drinks. If necessary, the produced aldosylfructoside can be isolated from the reaction mixture or the purified solution.
In respect to concentration, drying and pulverization, various conventional methods are employable in the present invention, for example evaporation and drying under reduced pressure, and spray-drying.
The sweetness of the sweetener containing aldosylfructoside thus obtained is equal to or slightly higher tha~ that of the substrate solution, and much milder on the basis of dry solid. In addition, the present sugar mixture containing aldosylfructoside offers an advantage that it does not cause crystallization no matter how high the concentration is or how long the period of storage is, although the composition of the substrate solution is much liable to crystallization. Furthermore, the sugar mixture has a sufficient viscosity and moisture-holding capacity, which can be used for imparting a sufficient viscosity, moisture retension capacity, gloss and body to foods and drinks, and improving their textures, as well as sweetening them.
For sweetening foods and drinks, the sweetener containing aldosylfructoside can be used alone or, if necessary, in combination with or in mixture with other sweeteners such as sucrose, glucose, maltose, corn syrup, isomerized sugar, honey, maple sugar, sorbitol, maltitol, lactitol, dihydrochalcone, L-asparatyl-L-phenylalanine methyl ester, saccharin, glycine, alanine, glycyr-rhizin and stevioside, fillers such as starch, dextrin and lactose, coloring matters, flavors and seasonings.
Particularly, the sweeteners containing aldosylfructoside differ from sucrose and can be used advantageously as a main- or sub-ingredient to produce foods and drinks because of their low-cariogenic and/or anti-caries properties.
Also, since the sweeteners containing aldosylfructoside are compatible with various sour, salty, delicious, astringent, or bitter-tasting substances, they can be used to sweeten foods and drinks in general, and favorite foods, and to improve their tastes: for example together with various seasonings such as soy, powder soy, mayonnaise, dressings, vinegars, sauces, catsups and curry roux, Japanese style confectioneries; western style confectioneries such as breads, biscuits, crackers, pies, puddings, butter creams, cookies, custard - 1141;~Z6 creams, waffles, sponge cakes, doughnuts, chocolates, chewing gums, caramels and candies; ice creams and sherbets; syrups; pastes such as flour pastes, peanut pastes and fruit pastes; preserved fruits and vegetables such as jams, marmalades and preserves; pickles and pickle products; meat products such as hams and sausages; fish products such as fish hams and fish sausages; various dainties, daily dishes; bottled foods such as those of fishes, meats, fruits and vegetables; canned foods; liquors such as Japanese Sake, wines, whiskies, brandies and alcohol drinks; drinks such as coffee, cocoa, juices, carbonated, lactic acid drinks and those containing lactic acid-producing microorganisms;
and convenient foods such as those of puddings, cakes, juices and coffee.
Also, the sweeteners prepared according to the present invention are usable for improving tastes of feeds and pet foods for domestic animals and fowls, honey bees, silkworms, fishes and other animals. In addition to the above-described uses, the sweeteners can be used for sweetening, improving and altering tastes of various favorites, cosmetics, drugs and medicines in solid, paste and liquid forms; for example tobaccoes, tooth pastes, lipsticks, lipcreams, medicines for internal administration, troches, drops containing liver oil, refrigerants, cachous and gargles.
As described above, the term foods and drinks as used throughout the SPECIFICATION means all foods and drinks in general, favorites, feeds, pet foods, cosmetics, drugs, medicines and all products which used orally, as well as sweetener, wherein the aldosylfructoside is used.
Any method can be employed as far as the aldosylfructoside is admixed in foods and drinks in the steps prior to final processing, for example mixing, ll~l;~Z~
kneading, soaking, scattering, applying and injecting. An aldosylfructoside content of about 1 % or more per foods and drinks d.s.b. is required when only inhibition of cariogenicity is desired.
EXPERIMENTs below describe the preparations and characteristics of aldosylfructosides.
EXPERIMENT
Preparation of levansucrase 1-1. Levansucrase of BaciZZus Sixty liters of a liquid medium, consistina of 3 w/v % defatted soybean,
Sucrose, a typical sweetener with sufficient sweetness and body, is consumed in large amount in a production of foods and drinks. Recentlv, it has been demonstrated that sweetened foods and drinks, particularly those sweetened with sucrose, cause very often dental caries. Dental caries is generally caused as follows; sucrose intaken is converted by oral bacteria into water-insoluble glucans such as dextran which adheres on tooth surface in layers, sugar intaken pass throu~h the layers and reach to the tooth surface where they are fermented anaerobically into organic acids, and the acids act on the enamel. Since sucrose is a major factor in causing dental caries, the realization of a process for producing foods and drinks using low-cariogenic sugars has been in great expectation.
According to the invention there is provided a process for producing food products, characterized in that the food products are prepared with, or added with aldosylfructoside which is obtained hy subjecting a 5 - 50 w/w% substrate solution, containing sucrose, raffinose or a mixture thereof, and one or more members selected from the group consisting of D-xylose, L-arabinose, D-galactose, D-mannose, xylobiose, cellobiose, maltose, isomaltosc, lactose, kojibiose, laminaribiose, nigerose, xylotriose, cellotriose, maltotriose, isomaltotriose, panose, isopanose, and partial hydrolysates of starch, dextran, pullulan, curdlan, pachyman, elsinan, glucomannan, cellulose and xylan, in the weight ratio of 1 : 50 - 5~ : 1, to the enzymatic action of levansucrase (E.C. 2.4.1.10) at a pH in the range _A_ cb/lJ , of 3 - 10 and a temperature in the range of 20 - 80C for 0.1 - 100 hours.
The present inventors investigated processes for producing low-cariogenic foods and drinks. The present invention is based on the discovery resulting from the efforts that aldosylfructoside obtalned by subjecting a substrate solution containing aldose, and sucrose and/or raffinose to the action of levansucrase (E.C. 2.4.1.10) has an appropriate sweetness and also low-cariogenic - la -cb/
ZG
and/or anti-caries properties which offer aldosylfructoside as a favorable sweetener for producing sweetened low-cariogenic foods and drinks.
Aldoses usable in the present invention are those which can be converted into aldosylfructosides excluding sucrose and raffinose by transfructosylation with levansuc~ase. Namely, aldosyl mono-, di- and tri-saccharides excluding glucose and melibiose are preferable: for example one or more members of a group comprising D-xylose, L-arabinose, D-galactose, D-mannose, xylobiose, cellobiose, maltose, isomaltose, lactose, kojibiose, laminaribiose, nigerose, xylotriose, cellotriose, maltotriose, isomaltotriose, panose and isopanose.
In addition, sugar mixtures, D.E. about 10 to 70, prepared by partial hydrolysis of polysaccharides such as starch, dextran, pullulan, curdlan9 pachyman, elsinan, glucomannan, cellulose and xylan, are usable in the present invention.
Substrate solution referred in the present invention is an aqueous solution containing aldose which acts as an acceptor for fructose residue, and sucrose and/or raffinose which act as a donor of fructose residue, upon transfructosylation reaction with levansucrase.
The mole ratio of acceptor vs. donor in the range of about 1 : 50 to 50 : 1, and substrate concentration in the range of about 5 to 50 w/w %, are preferable.
Levansucrase (E.C. 2.4.1.10) employable in the present invention is that which forms non-reducing aldosylfructosides when allowed to act on the above-described substrate solution containing aldose, and sucrose and/or raffinose, to transfer the fructosyl residues of sucrose and/or raffinose to the reducing 1141~Z6 Cl-sites of the aldoses. For example, levansucrase from Actinomyces v~scosus, Ae~obacter ~ev~icum, Acetobacter svboxydens, Boc~us Zichenifo~mis, Bac~us subti~is, G~uconobacter oxydens~ Streptococcus mutons, Streptococcus sa~ivaZius and other microorganisms is advantageously usable in the present invention.
In order to prepare levansucrase from the microorganisms, submerged culture is usually employed. The culture broth can be used without pre-treatment, but its supernatant or filtrate is usually used after removing water-insoluble substances by centrifugation or filtration. In some cases, the microbes can be used intact as an enzyme preparation without extraction, as well as levansucrase extracted from the cells by conventional methods. The levansucrase thus obtained is, if desired, purified further according to conventional method. In addition, immobilized levansucrases are also usable in continuous or batch-wise operation.
In the production of aldosylfructoside, a substrate solution containing aldose, and sucrose and/or raffinose is added with levansucrase to effect the enzymatic transfructosylation react;on. In this case, any reaction temperature and pH can be applied so far as the levansucrase acts on the substrate to produce corresponding aldosylfructoside; generally, the pH range of 3 to 10 and the temperature range of about 20 to 80CC are preferable. Levansucrase is used in the range of about 0.01 to 1,000 units per 9 sucrose and/or raffinose as defined by the following assay method, and the reaction is performed usually for about 0.1 to 100 hours.
Assay of levansucrase activitY
Two ml of a reaction mixture containing 10 w/v % sucrose, 50 mM
phosphate buffer, pH 7.0, and levansucrase is incubated at 30C for 30 minutes. The reaction mixture is then heated to inactivate the enzyme and the amount of the glucose released is determined by the glucose-oxidase method. One unit of levansucrase activity is defined as the amount of enzyme that produce 1 ~ mole of glucose per minute under the above conditions.
The reaction mixture containing aldosylfructoside can be, if desired, treated further with other enzymes such as glucose isomerase which isomerizes the glucose released in the reaction mixture into fructose and increases the sweetness of the product.
The transfructosylation reaction is usually suspended by heating and the reaction mixture i 5 filtrated. The filtrate is decolorized with activated carbon and deionized with ion exchangers, for example those of H-form and OH-form. The purified solution containing aldosylfructoside is concentrated into syrup, or dried and pulverized into powder for use in the production of foods and drinks. If necessary, the produced aldosylfructoside can be isolated from the reaction mixture or the purified solution.
In respect to concentration, drying and pulverization, various conventional methods are employable in the present invention, for example evaporation and drying under reduced pressure, and spray-drying.
The sweetness of the sweetener containing aldosylfructoside thus obtained is equal to or slightly higher tha~ that of the substrate solution, and much milder on the basis of dry solid. In addition, the present sugar mixture containing aldosylfructoside offers an advantage that it does not cause crystallization no matter how high the concentration is or how long the period of storage is, although the composition of the substrate solution is much liable to crystallization. Furthermore, the sugar mixture has a sufficient viscosity and moisture-holding capacity, which can be used for imparting a sufficient viscosity, moisture retension capacity, gloss and body to foods and drinks, and improving their textures, as well as sweetening them.
For sweetening foods and drinks, the sweetener containing aldosylfructoside can be used alone or, if necessary, in combination with or in mixture with other sweeteners such as sucrose, glucose, maltose, corn syrup, isomerized sugar, honey, maple sugar, sorbitol, maltitol, lactitol, dihydrochalcone, L-asparatyl-L-phenylalanine methyl ester, saccharin, glycine, alanine, glycyr-rhizin and stevioside, fillers such as starch, dextrin and lactose, coloring matters, flavors and seasonings.
Particularly, the sweeteners containing aldosylfructoside differ from sucrose and can be used advantageously as a main- or sub-ingredient to produce foods and drinks because of their low-cariogenic and/or anti-caries properties.
Also, since the sweeteners containing aldosylfructoside are compatible with various sour, salty, delicious, astringent, or bitter-tasting substances, they can be used to sweeten foods and drinks in general, and favorite foods, and to improve their tastes: for example together with various seasonings such as soy, powder soy, mayonnaise, dressings, vinegars, sauces, catsups and curry roux, Japanese style confectioneries; western style confectioneries such as breads, biscuits, crackers, pies, puddings, butter creams, cookies, custard - 1141;~Z6 creams, waffles, sponge cakes, doughnuts, chocolates, chewing gums, caramels and candies; ice creams and sherbets; syrups; pastes such as flour pastes, peanut pastes and fruit pastes; preserved fruits and vegetables such as jams, marmalades and preserves; pickles and pickle products; meat products such as hams and sausages; fish products such as fish hams and fish sausages; various dainties, daily dishes; bottled foods such as those of fishes, meats, fruits and vegetables; canned foods; liquors such as Japanese Sake, wines, whiskies, brandies and alcohol drinks; drinks such as coffee, cocoa, juices, carbonated, lactic acid drinks and those containing lactic acid-producing microorganisms;
and convenient foods such as those of puddings, cakes, juices and coffee.
Also, the sweeteners prepared according to the present invention are usable for improving tastes of feeds and pet foods for domestic animals and fowls, honey bees, silkworms, fishes and other animals. In addition to the above-described uses, the sweeteners can be used for sweetening, improving and altering tastes of various favorites, cosmetics, drugs and medicines in solid, paste and liquid forms; for example tobaccoes, tooth pastes, lipsticks, lipcreams, medicines for internal administration, troches, drops containing liver oil, refrigerants, cachous and gargles.
As described above, the term foods and drinks as used throughout the SPECIFICATION means all foods and drinks in general, favorites, feeds, pet foods, cosmetics, drugs, medicines and all products which used orally, as well as sweetener, wherein the aldosylfructoside is used.
Any method can be employed as far as the aldosylfructoside is admixed in foods and drinks in the steps prior to final processing, for example mixing, ll~l;~Z~
kneading, soaking, scattering, applying and injecting. An aldosylfructoside content of about 1 % or more per foods and drinks d.s.b. is required when only inhibition of cariogenicity is desired.
EXPERIMENTs below describe the preparations and characteristics of aldosylfructosides.
EXPERIMENT
Preparation of levansucrase 1-1. Levansucrase of BaciZZus Sixty liters of a liquid medium, consistina of 3 w/v % defatted soybean,
2 w/v % glucose, 4 w/v % sucrose, 0.6 w/v % (NH4)2HP04, 0.03 w/v % MgS04-7H20, 0.02 w/v % KCl, 0.02 w/v % calsium acetate, 0.001 w/v % MnS04 4H20 and water, was adjusted to pH 7.0, sterilized at 120C for 20 minutes, inoculated with BaciZZus subtiZis ATCC 6051 after cooling, and was cultured at 37C for 3 days under agitating under aerobic conditions. After completion of the cultivation, the culture broth was centrifuged to obtain a supernatant. Then the supernatant was added with equivolume of cold ethanol, and the formed precipitate was centrifuged, collected, dissolved in a 20 mM acetate buffer containing 1 mM
calsium chloride, pH 5.0, and dialyzed against a fresh buffer of the same composition overnight. The resultant was centrifuged and the obtained super-natant was applied on a DEAE-cellulose-packed column to adsorb the levansucrase, and the column was washed with a fresh buffer of the same composition. The adsorbed enzyme was eluted with a fresh buffer of the same composition except that it cdntained 1 M NaCl additionally. The eluate was saturated to 90 %
saturation with ammonium sulfate and the formed precipitate was centrifuged, collected and dissolved in 500 ml of the same buffer. The levansucrase activity of the solution was about 120 units per ml.
1-2. Levansucrase of Aerobacter Fifty liters of a liquid medium, consisting of 0.2 w/v % pepton, 1 w/v %
sucrose, 0.7 w/v % K2HPO4, 0.3 w/v % KH2PO4, 0.05 w/v % MgSO4 7H20, 0.05 w/v %
KCl, 0.001 w/v % FeSO4 7H20 and water, was sterilized at 120C for 20 minutes, and inoculated with Aerobacter Zevc~nicurn ATCC 15552, and the mixture was cultured at 30~C for 3 days under agitating under aerobic conditions. The culture broth was treated similarly as described in EXPERIMENT 1-1 and 600 ml of a levansucrase solution was obtained. The activity of the solution was about 150 units per ml.
Preparation of aldosyl fructosi de To a substrate solution, pH 6.0, containing 0.1 M sucrose as donor and 0.5 M of a member of a group comprising L-arabinose, D-xylose, D-mannose, D-galactose, maltose, isomaltose, lactose, cellobiose and maltotriose, was added a levansucrase, 2 units per 9 sucrose, prepared by the method as described in EXPERIMENT 1-1, and the resulting solution was incubated at 40C for 44 hours to effect the enzymatic transfructosylation reaction.
Analysis of the reaction mixtures according to the oligosaccharide map method described by J.H. Pazur and S. Okada, "The Journal of Biological Chemistry ', vol.241, pp.4146-4151 (1966), demonstrated in each case the transfer of the fructose residue of sucrose to the reducing group of acceptor aldose and formation of corresponding non-reducing aldosylfructoside;
arabinosyl fructosi de, xylosylfructoside, mannosyl fructosi de, galactosyl fructoside, g _ maltosylfructoside, isomaltosylfructoside, lactosylfructoside, cellobiosyl-fructoside and maltotriosylfructoside.
Isolation of the aldosylfructosides from the reaction mixtures was performed by conventional method with activated carbon-packed column: passing the reaction ~ixture through the column to adsorb all the sugars, eluting and fractiona~ing the adsorbed sugars with aqueous alcohol on concentration gradient, and then concentrating and drying the eluate containing aldosyl-fructoside into powder.
Hydrolysis of the aldosylfructosides by a commercial yeast ~-fructo-furanosidase (E.C. 3.2.1.26) demonstrated that every aldosylfructoside was hydrolyzed into fructose and the starting aldose. Therefore, it was shown that the present aldosylfructosides were aldosyl-~-fructofuranosides. The yields of the respective aldosylfructosides of high purity were about 15 %
on a mole basis of material aldoses. The aldosylfructosides were mildly sweet.
Low-cariogenic and anti-caries properties of aldosylfructoside In this EXPERIMENT with the aldosylfructosides prepared by the method as described in EXPERIMENT 2, the formation of lactic acid and water-insoluble glucan by an oral bacterium was determined.
calsium chloride, pH 5.0, and dialyzed against a fresh buffer of the same composition overnight. The resultant was centrifuged and the obtained super-natant was applied on a DEAE-cellulose-packed column to adsorb the levansucrase, and the column was washed with a fresh buffer of the same composition. The adsorbed enzyme was eluted with a fresh buffer of the same composition except that it cdntained 1 M NaCl additionally. The eluate was saturated to 90 %
saturation with ammonium sulfate and the formed precipitate was centrifuged, collected and dissolved in 500 ml of the same buffer. The levansucrase activity of the solution was about 120 units per ml.
1-2. Levansucrase of Aerobacter Fifty liters of a liquid medium, consisting of 0.2 w/v % pepton, 1 w/v %
sucrose, 0.7 w/v % K2HPO4, 0.3 w/v % KH2PO4, 0.05 w/v % MgSO4 7H20, 0.05 w/v %
KCl, 0.001 w/v % FeSO4 7H20 and water, was sterilized at 120C for 20 minutes, and inoculated with Aerobacter Zevc~nicurn ATCC 15552, and the mixture was cultured at 30~C for 3 days under agitating under aerobic conditions. The culture broth was treated similarly as described in EXPERIMENT 1-1 and 600 ml of a levansucrase solution was obtained. The activity of the solution was about 150 units per ml.
Preparation of aldosyl fructosi de To a substrate solution, pH 6.0, containing 0.1 M sucrose as donor and 0.5 M of a member of a group comprising L-arabinose, D-xylose, D-mannose, D-galactose, maltose, isomaltose, lactose, cellobiose and maltotriose, was added a levansucrase, 2 units per 9 sucrose, prepared by the method as described in EXPERIMENT 1-1, and the resulting solution was incubated at 40C for 44 hours to effect the enzymatic transfructosylation reaction.
Analysis of the reaction mixtures according to the oligosaccharide map method described by J.H. Pazur and S. Okada, "The Journal of Biological Chemistry ', vol.241, pp.4146-4151 (1966), demonstrated in each case the transfer of the fructose residue of sucrose to the reducing group of acceptor aldose and formation of corresponding non-reducing aldosylfructoside;
arabinosyl fructosi de, xylosylfructoside, mannosyl fructosi de, galactosyl fructoside, g _ maltosylfructoside, isomaltosylfructoside, lactosylfructoside, cellobiosyl-fructoside and maltotriosylfructoside.
Isolation of the aldosylfructosides from the reaction mixtures was performed by conventional method with activated carbon-packed column: passing the reaction ~ixture through the column to adsorb all the sugars, eluting and fractiona~ing the adsorbed sugars with aqueous alcohol on concentration gradient, and then concentrating and drying the eluate containing aldosyl-fructoside into powder.
Hydrolysis of the aldosylfructosides by a commercial yeast ~-fructo-furanosidase (E.C. 3.2.1.26) demonstrated that every aldosylfructoside was hydrolyzed into fructose and the starting aldose. Therefore, it was shown that the present aldosylfructosides were aldosyl-~-fructofuranosides. The yields of the respective aldosylfructosides of high purity were about 15 %
on a mole basis of material aldoses. The aldosylfructosides were mildly sweet.
Low-cariogenic and anti-caries properties of aldosylfructoside In this EXPERIMENT with the aldosylfructosides prepared by the method as described in EXPERIMENT 2, the formation of lactic acid and water-insoluble glucan by an oral bacterium was determined.
3-1. Formation of lactic acid 1.8 ml Aliquots of a mixture, containina a 0.1 M phosphate buffer, pH 6.8, 5 mM MgCl2 and a cell suspension of Streptococc~s mutans 6715, about 2.5 mg d.s.b., were shaken at 35~C for 5 minutes and the reaction was started by an addition of 0.2 ml of a 0.1 M aqueous aldosylfructoside solution to the mixture aliquots, and 20 minutes later, was stopped by an addition of 0.2 ml of a ~5 w/v ~ metaphosphoric acid soluiion. Then the reaction mixture aliquots were centrifuged and the amounts of the lactic acid formed in the supernatants were assayed according to the lactate dehydrogenase method. Control experiment was carried out similarly, except that the aldosylfructoside was replaced by sucrose. The results are shown in the TABLE, wherein the amounts of the formed lactic acid are expressed in percentages of that in the control.
3-2. Formation of water-insoluble glucan A commercial heart infusion medium containing bovine heart extract, pepton and NaCl was dissolved to give a concentration of about 2.5 w/v %, and added with respective aldosylfructosides prepared by the method as decsribed in EXPERIMENT 2 to give aldosylfructoside concentrations of about 2 w/v %.
Streptococcus mutons 6715 was inoculated in 4 ml of the above mixtures and the resulting mixtures were incubated at 37C for 16 hours. After cultivation, the cultures were centrifuged. The formed precipitates were added respectively with 4 ml of a 0.5 N NaOH, and incubated at 37C for an additional one hour to dissolve the water-insoluble glucan. The resulting mixtures were centri~fuged again and the amounts of the water-insoluble glucan formed in the supernatants were assayed by the phenol-sulfuric acid method. Control 11'~1;~;;~6 experiment was carried out similarly, except that the aldosylfructoside was replaced by sucrose. The results are shown in the TABLE, wherein the amounts of the formed water-insoluble glucan are expressed in percentages of that in the control.
3-3. Inhibition of pH-decrease and formation of water-insoluble glucan from sucrose Streptococcus mutans 6715 was inoculated similarly as in the EXPERIMENT
3-2 at 37C for 16 hours on a medium in which 80 % of the aldosylfructosides were replaced by sucrose. The pH-levels of the mixtures were monitored with a pH-meter and the amounts of the formed water-insoluble glucan were assayed similarly as described in EXPERIMENT 3-2. Control experiment was carried out similarly, except that sucrose was used in the amount of 2 w/v % and the aldosylfructosides were omitted. The results are shown in the TABLE, wherein the amounts of the formed water-insoluble glucan are expressed in percentages of that in the control.
As obvious from the results shown in the TABLE, the results of EXPERIMENTs 3-1 and 3-2 demonstrated that aldosylfructosides led to much less formation of lactic acid and water-insoluble glucan than the control, and the results of EXPERIMENT 3-3 demonstrated that use of the aldosylfructoside led to extreme inhibition on formation of water-insoluble glucan and pH-decrease from sucrose.
Thus, from the above-described results, it can be concluded that aldosylfructosides are suitable as a low-cariogenic and/or anti-caries sweetener.
Several embodiments according to the present invention are described below.
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ll~l'~Z6 E X A M P L E
EXAMPLE 1. Sweetener A solution prepared by dissolving 3 kg of sucrose and 1 kg of xylose in 10 1 of water was added with a levansucrase, 5 units per 9 sucrose, prepared by the method as described in EXPERIMENT 1-1, and incubated at 40C and pH 6.0 for 16 hours to effect the enzymatic transfructosylation reaction. Then the enzyme was inactivated by heating and the reaction mixture was filtrated. The filtrate was decolorized with activated carbon and deionized with ion exchangersof H-fGrm and OH-form according to conventional methods, and concentrated to give a syrup having a water content of about 20 w/w %, relatively low viscosity and high sweetness. The yield of the syrup sw_etener was about 95 % against material sugars d.s.b.
Because the syrup, xylosylfructoside content about 30 w/w % d.s.b., inhibits extremely the occurrence of dental caries when used alone and also in mixtures with sucrose, it is a superior low-cariogenic and/or anti-caries sweetener.
EXAMPLE 2. Sweetener A solution prepared by dissolving 40 kg of sucrose and 20 kg of maltose in 100 1 of water was added with a levansucrase, 2 units per 9 sucrose, prepared by the method as described in EXPERIFl~NT 1-1, and incubated at 40C and pH 6.0 for 44 hours to effect the reaction. The reaction mixture was purified and concentrated similarly as described in EXAMPLE 1 to give a water content of about 30 w/w %, and then spray-dried into white powder having a relatively high sweetness. The yield was about 90 % against material sugars d.s.b.
The sweetener, maltosylfructoside content about 30 % d.s.b., is feasible as a low-cariogenic sweetener.
EXAMPLE 3. Sweetener A solution prepared by dissolving 1 kg of sucrose and 5 kg of a high maltose syrup, water content 25 w/w % and D.E. about 60, in 7 1 of water was added with a levansucrase, 10 units per 9 sucrose, prepared by the method as described in EXPERIMENT 1-2, and incubated at 35C and pH 5.5 for 14 hours to effect the enzymatic reaction. Then the reaction mixture was purified and concentrated similarly as described in EXAMPLE 1 to give a syrup having a relatively high sweetness and a water content of about 20 w/w %. The yield was about 93 %
against material sugars d.s.b.
The syrup, containing about 10 % of aldosylfructosides excluding sucrose d.s.b., is suitable as a low-cariogenic sweetener and for imparting a proper moisture retension capacity and gloss to foods and drinks.
EXAMPLE 4. Sweetener A solution prepared by dissolving 1 kg of sucrose and 2 kg of a partially-hydrolyzed powder dextran, D.E. about 30, in 4 1 of water was added with a levansucrase, 2 units per 9 sucrose, prepared by the method as described in ll~l;~Z6 1-2, and incubated at 35C and pH 5.5 for 40 hours to effect the enzymatic reaction. The reaction mixture was purified and concentrated similarly as described in EXAMPLE 1 to give a syrup sweetener having a water content of about 20 w/w % and a moderate sweetness. The yield was about 96 ~0 against starting materials d.s.b.
Because the syrup, the content of aldosylfructosides excluding sucrose about 25 % d.s.b., inhibits extremely the occurrence of dental caries when used alone or in mixtures with sucrose, it is an excellent low-cariogenic and/or anti-caries sweetener. In addition, the sweetener is suitable for imparting a proper sweetness, moisture retension capacity and gloss to foods and drinks.
EXAMPLE 5. Sweetener A solution was prepared by dissolving 50 kg of sucrose and 10 kg of lactose in 70 1 of water. The solution was added with a levansucrase, 1 unit per 9 sucrose, prepared by the method as described in EXPERIMENT 1-2, incubated at 40C and pH 6.0 for 40 hours to effect the enzymatic reaction and then incubated at 60~C for an additional 5 hours after addition of a commercial glucose isomerase, 20 units per 9 sucrose. The reaction mixture was purified and concentrated similarly as described in EXAMPLE 2, and then spray-dried to give a powder sweetener having a relatively high sweetness. The yield was about 92 % against material sugars d.s.b.
The powder product, containing about 20 % of lactosylfructoside d.s.b., is suitable as a low-cariogenic sweetener.
ll~l;~Z6 EXAMPLE 6. Sweetener A solution prepared by dissolving 2 kg of raffinose and 1 kg of a powder maltodextrin, D.E. about 20, in 4 1 of water was added with a levansucrase, 3 units per 9 raffinose, prepared by the method as described in EXPERIMENT
1-1, and incubated at 40C and pH 6.0 for 20 hours to effect the enzymatic reaction. Then the reaction mixture was purified and concentrated similarly as described in EXAMPLE 1 to give a syrup sweetener having a low sweetness, high viscosity and water content of about 20 w/w ~. The yield was about 94 %
against starting materials d.s.b.
The syrup thus obtained, the content of aldosylfructosides excluding raffinose about 25 % d.s.b., is suitable as a low-cariogenic sweetener and can be used to impart a proper viscosity, moisture retension capacity and gloss to foods including drinks.
EXAMPLE 7. Sweetener A syrup sweetener mixture was prepared by dissolving 250 9 of a powder sweetener as prepared by the method describcd in EXAMPLE 2 in 1 kg of a hydrogenated maltose syrup, water content about 25 w/w %.
The mixture has the same sweetness as sucrose, and therefore is suitable as a diet sweetener for diabetics and obeses as well as a low-cariogenic sweetener. Also, the sweetener offers an advantage that foods can be prepared with much less browning when the sweetener is used because it tolerates heating and is thus less liable to colorizing foods. In addition, the l.l~ Z6 the sweetener is also suitable for imparting a sufficient moisture retension capacity and gloss to foods including drinks.
EXAMPLE 8. Hard candy Ten kg of a syrup prepared by the method as described in EXAMPLE 7 was heated and evaporated under reduced pressure to a moisture content of below 2 w/w %, mixed with 100 9 of citric acid, small amounts of a lemon flavour and a coloring matter and shaped into hard candy according to conventional method.
The hard candy is low-cariogenic.
EXAMPLE 9. Chewing gum Two kg of a gum base was heated and softened, and mixed with 7 kg of a powder sweetener prepared by the method as described in EXAMPLE 5, small amounts of peppermint and coloring matter. Then the mixture was kneaded with a roller and shaped into chewing gums according to conventional method.
The chewing gum has a good taste and texture, and low-cariogenicity.
EXAMPLE 10. Chocolate Forty kg of cacao paste, 10 kg of cacao butter, 15 kg of a powder sweetener prepared by the method as described in EXAMPLE 2 and 20 kg of a whole milk were mixed and then passed through a refiner to reduce the solid particles.
Then the mixture was added with 500 9 of lecithin and kneaded at 50~C for 2 ..
11'~ 6 dadys in a conche. The resulting mixture was moulded and solidified according to conventional method.
The low-cariogenic chocolate thus obtained is less liable to fat and sugar blooms, and also very tasty and appealing to the palate.
EXAMPLE 1l; Lactic acid drink Ten kg of skim milk was pasteurized by heating at 80C for 20 minutes and cooled to 40C. The milk was then added with 300 9 of a starter and fermented at a temperature of about 35 to 37C for 10 hours. The obtained mixture was homogenized, added with 7 kg of a syrup prepared by the method as described in EXAMPLE 3, and pasteurized by heating at a temperature of about 60 to 65C
while suppressing decomposition of the aldosylfructoside contents. The mixture was mixed with a small amount of flavour and bottled after cooling.
The drink is suitable as a low-cariogenic drink, and its sweetness and flavour are compatible with its sour taste.
EXAMPLE 12. Tsukudani (Japanese style preserved food boiled down in soy) After removing sand from 250 9 of tangle, the tangle was treated with an acid, cut into small squares according to conventional method and soaked in a solution comprising 212 ml of soy, 318 ml of amino acid solution and 100 9 of a syrup prepared by the method as described in EXAMPLE 4. The mixture was added additionally with 12 9 of sodim glutamate, 8 9 of caramel and 21 ml of Mirin (sweet Sake) while boiling the mixture. Then the mixture was boiled down ~ 2 6 to obtain Tsukudani.
The produc~ was an appetizing and appealing Tsukudani in colour and gloss as well as in flavour.
EXAMPLE 13. Pickled scallion-Five kg of fresh scallions were soaked in 2.5 1 of a 20 w/v ~ saline so1ution for 3 weeks according to conventional method and the solution was drained off. The salted scallions were pickled for one month in an acetic acid solution containing 80 9 of sodium chloride, 80 ml of glacial acetic acid and 2.0 1 of water. The pickled scallions thus obtained were then pickled in a fresh pickle solution consisting of 800 ml of vinegar, 200 ml of Mirin, 0.5 9 of stevioside, 10 9 of red pepper and 150 9 of a sweetener prepared by the method as described in EXAMPLE 6 for an additional 10 days to obtain tasty pickled scallions.
EXAMPLE 14. Tablet A 50 g of aspirin was admixed homogenously with 4 9 of cornstarch and 14 g of a powder sweetener prepared by the method as described in EXAMPLE 2. A
tabletting machine equipped with a pestle having a diameter of 12 mm and a curvature of 20 R was used to form each tablets, having a weight of 680 mg, a thickness of 5.25 mm and a hardness of 8 + 1 kg.
The tablets are less liable to crack and deformation during long period of storage and intaken easily because of their pleasant sweetness.
EXAMPLE 15. Tooth paste The below-described materials were mixed according to conventional method to prepare a tooth paste.
Formulation Dipotassium hydrogenphosphate 45 %
Pullulan 2.95 %
Sodium lauryl sulfate 1.5 %
Glycerin 20 %
Polyoxyethylene sorbitan laurate 0.5 %
Antiseptic a~ent 0.05 %
A liquid sweetener prepared by the 18 %
method as described in EXAMPLE 1 Water 12 %
The tooth paste is suitable for children owing to its appropriate sweetness.
3-2. Formation of water-insoluble glucan A commercial heart infusion medium containing bovine heart extract, pepton and NaCl was dissolved to give a concentration of about 2.5 w/v %, and added with respective aldosylfructosides prepared by the method as decsribed in EXPERIMENT 2 to give aldosylfructoside concentrations of about 2 w/v %.
Streptococcus mutons 6715 was inoculated in 4 ml of the above mixtures and the resulting mixtures were incubated at 37C for 16 hours. After cultivation, the cultures were centrifuged. The formed precipitates were added respectively with 4 ml of a 0.5 N NaOH, and incubated at 37C for an additional one hour to dissolve the water-insoluble glucan. The resulting mixtures were centri~fuged again and the amounts of the water-insoluble glucan formed in the supernatants were assayed by the phenol-sulfuric acid method. Control 11'~1;~;;~6 experiment was carried out similarly, except that the aldosylfructoside was replaced by sucrose. The results are shown in the TABLE, wherein the amounts of the formed water-insoluble glucan are expressed in percentages of that in the control.
3-3. Inhibition of pH-decrease and formation of water-insoluble glucan from sucrose Streptococcus mutans 6715 was inoculated similarly as in the EXPERIMENT
3-2 at 37C for 16 hours on a medium in which 80 % of the aldosylfructosides were replaced by sucrose. The pH-levels of the mixtures were monitored with a pH-meter and the amounts of the formed water-insoluble glucan were assayed similarly as described in EXPERIMENT 3-2. Control experiment was carried out similarly, except that sucrose was used in the amount of 2 w/v % and the aldosylfructosides were omitted. The results are shown in the TABLE, wherein the amounts of the formed water-insoluble glucan are expressed in percentages of that in the control.
As obvious from the results shown in the TABLE, the results of EXPERIMENTs 3-1 and 3-2 demonstrated that aldosylfructosides led to much less formation of lactic acid and water-insoluble glucan than the control, and the results of EXPERIMENT 3-3 demonstrated that use of the aldosylfructoside led to extreme inhibition on formation of water-insoluble glucan and pH-decrease from sucrose.
Thus, from the above-described results, it can be concluded that aldosylfructosides are suitable as a low-cariogenic and/or anti-caries sweetener.
Several embodiments according to the present invention are described below.
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ll~l'~Z6 E X A M P L E
EXAMPLE 1. Sweetener A solution prepared by dissolving 3 kg of sucrose and 1 kg of xylose in 10 1 of water was added with a levansucrase, 5 units per 9 sucrose, prepared by the method as described in EXPERIMENT 1-1, and incubated at 40C and pH 6.0 for 16 hours to effect the enzymatic transfructosylation reaction. Then the enzyme was inactivated by heating and the reaction mixture was filtrated. The filtrate was decolorized with activated carbon and deionized with ion exchangersof H-fGrm and OH-form according to conventional methods, and concentrated to give a syrup having a water content of about 20 w/w %, relatively low viscosity and high sweetness. The yield of the syrup sw_etener was about 95 % against material sugars d.s.b.
Because the syrup, xylosylfructoside content about 30 w/w % d.s.b., inhibits extremely the occurrence of dental caries when used alone and also in mixtures with sucrose, it is a superior low-cariogenic and/or anti-caries sweetener.
EXAMPLE 2. Sweetener A solution prepared by dissolving 40 kg of sucrose and 20 kg of maltose in 100 1 of water was added with a levansucrase, 2 units per 9 sucrose, prepared by the method as described in EXPERIFl~NT 1-1, and incubated at 40C and pH 6.0 for 44 hours to effect the reaction. The reaction mixture was purified and concentrated similarly as described in EXAMPLE 1 to give a water content of about 30 w/w %, and then spray-dried into white powder having a relatively high sweetness. The yield was about 90 % against material sugars d.s.b.
The sweetener, maltosylfructoside content about 30 % d.s.b., is feasible as a low-cariogenic sweetener.
EXAMPLE 3. Sweetener A solution prepared by dissolving 1 kg of sucrose and 5 kg of a high maltose syrup, water content 25 w/w % and D.E. about 60, in 7 1 of water was added with a levansucrase, 10 units per 9 sucrose, prepared by the method as described in EXPERIMENT 1-2, and incubated at 35C and pH 5.5 for 14 hours to effect the enzymatic reaction. Then the reaction mixture was purified and concentrated similarly as described in EXAMPLE 1 to give a syrup having a relatively high sweetness and a water content of about 20 w/w %. The yield was about 93 %
against material sugars d.s.b.
The syrup, containing about 10 % of aldosylfructosides excluding sucrose d.s.b., is suitable as a low-cariogenic sweetener and for imparting a proper moisture retension capacity and gloss to foods and drinks.
EXAMPLE 4. Sweetener A solution prepared by dissolving 1 kg of sucrose and 2 kg of a partially-hydrolyzed powder dextran, D.E. about 30, in 4 1 of water was added with a levansucrase, 2 units per 9 sucrose, prepared by the method as described in ll~l;~Z6 1-2, and incubated at 35C and pH 5.5 for 40 hours to effect the enzymatic reaction. The reaction mixture was purified and concentrated similarly as described in EXAMPLE 1 to give a syrup sweetener having a water content of about 20 w/w % and a moderate sweetness. The yield was about 96 ~0 against starting materials d.s.b.
Because the syrup, the content of aldosylfructosides excluding sucrose about 25 % d.s.b., inhibits extremely the occurrence of dental caries when used alone or in mixtures with sucrose, it is an excellent low-cariogenic and/or anti-caries sweetener. In addition, the sweetener is suitable for imparting a proper sweetness, moisture retension capacity and gloss to foods and drinks.
EXAMPLE 5. Sweetener A solution was prepared by dissolving 50 kg of sucrose and 10 kg of lactose in 70 1 of water. The solution was added with a levansucrase, 1 unit per 9 sucrose, prepared by the method as described in EXPERIMENT 1-2, incubated at 40C and pH 6.0 for 40 hours to effect the enzymatic reaction and then incubated at 60~C for an additional 5 hours after addition of a commercial glucose isomerase, 20 units per 9 sucrose. The reaction mixture was purified and concentrated similarly as described in EXAMPLE 2, and then spray-dried to give a powder sweetener having a relatively high sweetness. The yield was about 92 % against material sugars d.s.b.
The powder product, containing about 20 % of lactosylfructoside d.s.b., is suitable as a low-cariogenic sweetener.
ll~l;~Z6 EXAMPLE 6. Sweetener A solution prepared by dissolving 2 kg of raffinose and 1 kg of a powder maltodextrin, D.E. about 20, in 4 1 of water was added with a levansucrase, 3 units per 9 raffinose, prepared by the method as described in EXPERIMENT
1-1, and incubated at 40C and pH 6.0 for 20 hours to effect the enzymatic reaction. Then the reaction mixture was purified and concentrated similarly as described in EXAMPLE 1 to give a syrup sweetener having a low sweetness, high viscosity and water content of about 20 w/w ~. The yield was about 94 %
against starting materials d.s.b.
The syrup thus obtained, the content of aldosylfructosides excluding raffinose about 25 % d.s.b., is suitable as a low-cariogenic sweetener and can be used to impart a proper viscosity, moisture retension capacity and gloss to foods including drinks.
EXAMPLE 7. Sweetener A syrup sweetener mixture was prepared by dissolving 250 9 of a powder sweetener as prepared by the method describcd in EXAMPLE 2 in 1 kg of a hydrogenated maltose syrup, water content about 25 w/w %.
The mixture has the same sweetness as sucrose, and therefore is suitable as a diet sweetener for diabetics and obeses as well as a low-cariogenic sweetener. Also, the sweetener offers an advantage that foods can be prepared with much less browning when the sweetener is used because it tolerates heating and is thus less liable to colorizing foods. In addition, the l.l~ Z6 the sweetener is also suitable for imparting a sufficient moisture retension capacity and gloss to foods including drinks.
EXAMPLE 8. Hard candy Ten kg of a syrup prepared by the method as described in EXAMPLE 7 was heated and evaporated under reduced pressure to a moisture content of below 2 w/w %, mixed with 100 9 of citric acid, small amounts of a lemon flavour and a coloring matter and shaped into hard candy according to conventional method.
The hard candy is low-cariogenic.
EXAMPLE 9. Chewing gum Two kg of a gum base was heated and softened, and mixed with 7 kg of a powder sweetener prepared by the method as described in EXAMPLE 5, small amounts of peppermint and coloring matter. Then the mixture was kneaded with a roller and shaped into chewing gums according to conventional method.
The chewing gum has a good taste and texture, and low-cariogenicity.
EXAMPLE 10. Chocolate Forty kg of cacao paste, 10 kg of cacao butter, 15 kg of a powder sweetener prepared by the method as described in EXAMPLE 2 and 20 kg of a whole milk were mixed and then passed through a refiner to reduce the solid particles.
Then the mixture was added with 500 9 of lecithin and kneaded at 50~C for 2 ..
11'~ 6 dadys in a conche. The resulting mixture was moulded and solidified according to conventional method.
The low-cariogenic chocolate thus obtained is less liable to fat and sugar blooms, and also very tasty and appealing to the palate.
EXAMPLE 1l; Lactic acid drink Ten kg of skim milk was pasteurized by heating at 80C for 20 minutes and cooled to 40C. The milk was then added with 300 9 of a starter and fermented at a temperature of about 35 to 37C for 10 hours. The obtained mixture was homogenized, added with 7 kg of a syrup prepared by the method as described in EXAMPLE 3, and pasteurized by heating at a temperature of about 60 to 65C
while suppressing decomposition of the aldosylfructoside contents. The mixture was mixed with a small amount of flavour and bottled after cooling.
The drink is suitable as a low-cariogenic drink, and its sweetness and flavour are compatible with its sour taste.
EXAMPLE 12. Tsukudani (Japanese style preserved food boiled down in soy) After removing sand from 250 9 of tangle, the tangle was treated with an acid, cut into small squares according to conventional method and soaked in a solution comprising 212 ml of soy, 318 ml of amino acid solution and 100 9 of a syrup prepared by the method as described in EXAMPLE 4. The mixture was added additionally with 12 9 of sodim glutamate, 8 9 of caramel and 21 ml of Mirin (sweet Sake) while boiling the mixture. Then the mixture was boiled down ~ 2 6 to obtain Tsukudani.
The produc~ was an appetizing and appealing Tsukudani in colour and gloss as well as in flavour.
EXAMPLE 13. Pickled scallion-Five kg of fresh scallions were soaked in 2.5 1 of a 20 w/v ~ saline so1ution for 3 weeks according to conventional method and the solution was drained off. The salted scallions were pickled for one month in an acetic acid solution containing 80 9 of sodium chloride, 80 ml of glacial acetic acid and 2.0 1 of water. The pickled scallions thus obtained were then pickled in a fresh pickle solution consisting of 800 ml of vinegar, 200 ml of Mirin, 0.5 9 of stevioside, 10 9 of red pepper and 150 9 of a sweetener prepared by the method as described in EXAMPLE 6 for an additional 10 days to obtain tasty pickled scallions.
EXAMPLE 14. Tablet A 50 g of aspirin was admixed homogenously with 4 9 of cornstarch and 14 g of a powder sweetener prepared by the method as described in EXAMPLE 2. A
tabletting machine equipped with a pestle having a diameter of 12 mm and a curvature of 20 R was used to form each tablets, having a weight of 680 mg, a thickness of 5.25 mm and a hardness of 8 + 1 kg.
The tablets are less liable to crack and deformation during long period of storage and intaken easily because of their pleasant sweetness.
EXAMPLE 15. Tooth paste The below-described materials were mixed according to conventional method to prepare a tooth paste.
Formulation Dipotassium hydrogenphosphate 45 %
Pullulan 2.95 %
Sodium lauryl sulfate 1.5 %
Glycerin 20 %
Polyoxyethylene sorbitan laurate 0.5 %
Antiseptic a~ent 0.05 %
A liquid sweetener prepared by the 18 %
method as described in EXAMPLE 1 Water 12 %
The tooth paste is suitable for children owing to its appropriate sweetness.
Claims (18)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A process for producing food products, characterized in that said food products are prepared with, or added with aldosylfructoside which is obtained by subjecting a 5 - 50 w/w%
substrate solution, containing sucrose, raffinose or a mixture thereof, and one or more members selected from the group consisting of D-xylose, L-arabinose, D-galactose, D-mannose, xylobiose, cellobiose, maltose, isomaltose, lactose, kojibiose, laminaribiose, nigerose, xylotriose, cellotriose, maltotriose, isomaltotriose, panose, isopanose, and partial hydrolysates of starch, dextran, pullulan, curdlan, pachyman, elsinan, glucomannan, cellulose and xylan, in the weight ratio of 1 : 50 - 50 : 1, to the enzymatic action of levansucrase (E.C. 2.4.1.10) at a pH in the range of 3 - 10 and a temperature in the range of 20° - 80°C for 0.1 - 100 hours.
substrate solution, containing sucrose, raffinose or a mixture thereof, and one or more members selected from the group consisting of D-xylose, L-arabinose, D-galactose, D-mannose, xylobiose, cellobiose, maltose, isomaltose, lactose, kojibiose, laminaribiose, nigerose, xylotriose, cellotriose, maltotriose, isomaltotriose, panose, isopanose, and partial hydrolysates of starch, dextran, pullulan, curdlan, pachyman, elsinan, glucomannan, cellulose and xylan, in the weight ratio of 1 : 50 - 50 : 1, to the enzymatic action of levansucrase (E.C. 2.4.1.10) at a pH in the range of 3 - 10 and a temperature in the range of 20° - 80°C for 0.1 - 100 hours.
2. A process as set forth in claim 1, wherein said lavansucrase is obtained from a microorganism of species Actinomyces viscosus, Aerobacter lavanicum, Acetobacter suboxydans, Bacillus licheniformis, Bacillus subtilis, Gluconobacter oxydans, Streptococcus mutans or Streptococcus salivalius.
3. A process as set forth in claim 1, wherein D.E. of the hydrolysates are in the range of 10 - 70.
4. A process as set forth in claim 1, wherein the levansucrase is used in an amount of 0.01 - 1,000 units per g sucrose, raffinose or mixture thereof.
5. A process as set forth in claim 1, wherein the aldosylfructoside content in said food products is at least 1% on dry solid basis.
6. A process as set forth in claim 1, wherein said aldosylfructoside is arabinosylfructoside, xylosylfructoside, mannosylfructoside, galactosylfructoside, maltosylfructoside, isomaltosylfructoside, lactosylfructoside, cellobiosyl-fructoside, maltotriosylfructoside, or mixture thereof.
7. A process as set forth in claim 1, wherein said food products are sweeteners.
8. A process as set forth in claim 1, wherein said food products are confectioneries.
9. A process as set forth in claim 1, wherein said food products are bakery products.
10. A process as set forth in claim 1, wherein said food products are seasonings.
11. A process as set forth in claim 1, wherein said food products are frozen desserts.
12. A process as set forth in claim 1, wherein said food products are syrups.
13. A process as set forth in claim 1, wherein said food prouducts are preserved foods or preserves.
14. A process as set forth in claim 1, wherein said food products are pickles or pickled products.
15. A process as set forth in claim 1, wherein said food products are pastes.
16. A process as set forth in claim 1, wherein said food products are soft drinks.
17. A process as set forth in claim 1, wherein said food products are convenience foods.
18. A process as set forth in claim 1, wherein said food products are pet foods or feeds for domestic animals, fowls, honey bees, silk worms, or fish.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2505679A JPS55118369A (en) | 1979-03-06 | 1979-03-06 | Method of making beverage and food |
| JP25056/1979 | 1979-03-06 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1141226A true CA1141226A (en) | 1983-02-15 |
Family
ID=12155257
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000347168A Expired CA1141226A (en) | 1979-03-06 | 1980-03-06 | Low cariogenic food products |
Country Status (5)
| Country | Link |
|---|---|
| JP (1) | JPS55118369A (en) |
| CA (1) | CA1141226A (en) |
| DE (1) | DE3008668C2 (en) |
| FR (1) | FR2450876A1 (en) |
| GB (1) | GB2046757B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4518581A (en) * | 1981-11-02 | 1985-05-21 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Imparting low- or anti-cariogenic property to orally-usable products |
| GB8316790D0 (en) * | 1983-06-21 | 1983-07-27 | Tate & Lyle Plc | Chemical process |
| JPS6034134A (en) * | 1983-08-05 | 1985-02-21 | Meiji Seika Kaisha Ltd | Feed containing fructoligosaccharide and feeding of domestic animals therewith |
| EP0142230A3 (en) * | 1983-09-27 | 1986-06-11 | University Of Queensland | Conversion of sucrose to fructose and ethanol |
| CA1324529C (en) * | 1987-09-15 | 1993-11-23 | Tatsuhiko Kan | Liquid food comprising polydextrose and oligosaccharides |
| JP3778991B2 (en) * | 1996-03-04 | 2006-05-24 | 株式会社林原生物化学研究所 | Maltooligosyl turranose and maltooligosyl palatinose-containing saccharides, production method and use thereof |
| WO2012095746A2 (en) | 2011-01-11 | 2012-07-19 | Capsugel Belgium Nv | New hard capsules |
| JP2020516653A (en) | 2017-04-14 | 2020-06-11 | カプスゲル・ベルギウム・ナムローゼ・フェンノートシャップＣａｐｓｕｇｅｌ Ｂｅｌｇｉｕｍ ＮＶ | Pullulan capsule |
| WO2018189587A1 (en) | 2017-04-14 | 2018-10-18 | Capsugel Belgium Nv | Process for making pullulan |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1543167A (en) * | 1976-01-08 | 1979-03-28 | Tate & Lyle Ltd | Sweeteners |
-
1979
- 1979-03-06 JP JP2505679A patent/JPS55118369A/en active Granted
-
1980
- 1980-03-05 GB GB8007455A patent/GB2046757B/en not_active Expired
- 1980-03-06 CA CA000347168A patent/CA1141226A/en not_active Expired
- 1980-03-06 DE DE3008668A patent/DE3008668C2/en not_active Expired
- 1980-03-06 FR FR8005018A patent/FR2450876A1/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| DE3008668C2 (en) | 1985-01-03 |
| JPS5758905B2 (en) | 1982-12-11 |
| FR2450876A1 (en) | 1980-10-03 |
| FR2450876B1 (en) | 1985-05-10 |
| JPS55118369A (en) | 1980-09-11 |
| GB2046757A (en) | 1980-11-19 |
| GB2046757B (en) | 1983-04-20 |
| DE3008668A1 (en) | 1980-09-18 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKEX | Expiry |