KR101916330B1 - Composition for preventing or relieving hangover including allulose - Google Patents

Abstract

The present invention provides a composition for prevention or elimination of hangover comprising at least one member selected from red ginseng extract or hinoki extract and at least one member selected from allulose or allylose-containing complex saccharides. The allulose or allylose-containing complex saccharide, which is a component of the composition for preventing or eliminating a hangover according to the present invention, acts as a booster for red ginseng extract or Hovenia dulcis extract, It can increase the activity of aldehyde dehydrogenase (ALDH) and rapidly oxidize acetaldehyde. Therefore, the composition for preventing or eliminating a hangover according to the present invention exhibits a superior effect when using only red ginseng extract or Hovenia dulcis extract or using other saccharides together with red ginseng extract or Hovenia dulcis extract.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or relieving hangovers,

The present invention relates to a composition for preventing or eliminating a hangover, and more particularly, to a composition for preventing or eliminating a hangover which can rapidly oxidize acetaldehyde by increasing the activity of an aldehyde dehydrogenase (ALDH).

Hangover refers to headache, diarrhea, anorexia, nausea, vomiting, chills, and cold sweating that occur after drinking alcohol. Objective symptoms include cognition, decreased exercise capacity, hematological changes, and changes in hormones. The cause of hangover is known as dehydration, toxicity of alcohol and alcohol metabolites (acetaldehyde, formaldehyde, acetone, etc.), nutrient deficiency (blood sugar, vitamin and mineral deficiency) Hangover after drinking alcohol is not only toxicity caused by alcohol itself, but alcohol is transformed into harmful substance in human body during metabolic process and acts as harmful substance to digestive organs including brain and liver. The degree of hangover is very different according to the variation of individuals due to heredity, general condition (nutrition, exercise, dehydration, health).

After drinking alcohol, alcohol is absorbed mainly from the stomach and small intestine, is transferred to the liver and metabolized. Approximately 90% of alcohol reached in the liver is oxidized to acetaldehyde by alcohol dehydrogenase (ADH) and acetaldehyde is oxidized by aldehyde dehydrogenase (ALDH) to acetic acid ). The remaining about 10% of the alcohol is metabolized to acetaldehyde by catalase. Since acetaldehyde, a primary metabolite of alcohol, is a highly reactive compound, it reacts with various other substances in the body to increase toxicity and necrosis of hepatocytes, changes in microvessels, changes in structure and function of hepatocyte mitochondria and lipid peroxidation . That is, acetaldehyde generates acetaldehyde-protein adducts, which induce an immune response, thus causing liver damage. In addition, acetaldehyde is involved in hepatotoxicity by producing lipid peroxides. In addition, chronic alcohol intake inhibits the oxidation of fatty acids and increases the concentration of triglyceride (TG) in the blood, which causes the liver to progress to fatty liver. In addition, acetaldehyde inhibits the production of glutathione, which causes antioxidant effects by causing metabolic abnormalities of methionine. The free radicals produced in the metabolism of alcohol are mainly removed by antioxidant enzymes such as catalase, superoxide dismutase (SOD), and glutathione peroxidase The generation of free radicals leads to liver damage, that is, apoptosis or necrosis of hepatocytes, beyond the antioxidant defense period. Acetaldehyde, the first metabolite of alcohol accumulated in the liver, is a highly active compound that causes sweat, rapid pulse, skin flushing, nausea, vomiting, For this reason, most researchers believe that acetaldehyde, an alcohol degradation product, is a major cause of acute alcohol hangover. Therefore, in order to effectively solve the alcoholic hangover in a short time, development of a material having an ability to decompose acetaldehyde, which can effectively reduce the concentration of acetaldehyde, is required.

Hangover symptoms start within hours of drinking and last for almost a day, and the experience of a particular hangover is highly dependent on personal preference and situational and type and amount of alcohol consumed. Symptoms of physical hangover include headache, fatigue, light and sound sensitivity, myalgia, redness of the eyes, thirst, nausea, vomiting and stomach pain. Hangover causes fast heartbeat, excitement, elevated blood pressure and sweating. Related mental symptoms of hangover include sleep deprivation, sleeping location changes, decreased attention, decreased concentration, depression, dizziness, anxiety, and irritability. Although various synthetic medicines have been developed in order to solve hangover after drinking, much attention has been paid to the development of safer natural drugs, health supplements, and health drinks due to their toxicity or side effects.

Regarding the material derived from natural materials for hangover removal, Korean Patent No. 10-1615199 discloses that 30 to 50 parts by weight of Hovenia dulcis extract, 10 to 15 parts by weight of a dandruff liquid of dandelion outpour, 15 to 30 parts by weight of asparagine, 10-15 parts by weight of a polyp extract and 15-30 parts by weight of a lactic acid bacterium. Korean Patent No. 10-0771525 also discloses a composition for use as a hangover decoy containing an extract of Hovenia dulcis, Injinho, Puerariae, Dermis, Creation and Licorice as an active ingredient, which is extracted with water, C1 to C3 lower alcohol or a mixture thereof Lt; / RTI > Korean Patent No. 10-1668447 also discloses that red ginseng extract; And extracts of mixed herbal medicines consisting of marine oil, omija and algae; And a red ginseng chip as an active ingredient. Also, Korean Patent No. 10-1324431 discloses a composition for hangover decomposing which comprises red ginseng extract, microbial soybean fermentation extract, mulberry extract, aglymoni extract, and upper leaf extract as effective ingredients. Korean Patent Laid-Open No. 10-2009-0116623 discloses a hangover-eliminating beverage comprising ginseng fruit extract, ginseng vase extract, hornbug fruit extract and saccharide.

It is an object of the present invention to provide a composition for prevention or elimination of hangover which can rapidly oxidize acetaldehyde.

The inventors of the present invention found that when combined sugars containing allulose or alulose are used in combination with red ginseng extract or hodgkin's extract, the activity of aldehyde dehydrogenase (ALDH), which is exerted in red ginseng extract or hinoki extract, Can be oxidized rapidly, and the present invention has been completed.

In order to achieve the above object, an example of the present invention provides a composition for preventing or eliminating a hangover comprising at least one selected from red ginseng extract or Hovenia dulcis extract and alulose.

In order to achieve the above object, another embodiment of the present invention is a composition comprising at least one selected from red ginseng extract or Hovenia dulcis extract and a complex saccharide, 80 to 99.9% by weight of losse and 0.1 to 20% by weight of sucrose.

The allulose or allylose-containing complex saccharide, which is a component of the composition for preventing or eliminating a hangover according to the present invention, acts as a booster for red ginseng extract or Hovenia dulcis extract, It can increase the activity of aldehyde dehydrogenase (ALDH) and rapidly oxidize acetaldehyde. Therefore, the composition for preventing or eliminating a hangover according to the present invention exhibits a superior effect when using only red ginseng extract or Hovenia dulcis extract or using other saccharides together with red ginseng extract or Hovenia dulcis extract.

The present invention relates to a composition for prevention or elimination of hangover which can rapidly oxidize acetaldehyde by increasing the activity of aldehyde dehydrogenase (ALDH).

The composition for preventing or eliminating a hangover according to one embodiment of the present invention includes at least one selected from red ginseng extract or Hovenia dulcis extract and alulose. The composition for preventing or eliminating a hangover according to one embodiment of the present invention preferably comprises 100 to 800 parts by weight of alulose based on 100 parts by weight of the red ginseng extract and more preferably 100 to 800 parts by weight of alulose 250 To 600 parts by weight. In addition, the composition for preventing or eliminating a hangover according to one embodiment of the present invention is preferably a hovenia dulcis extract according to 100 parts by weight of the extract, more preferably 400 to 1500 parts by weight of aloe vera, dulcis ) extract in an amount of 600 to 1400 parts by weight based on 100 parts by weight of the extract. In addition, the composition for preventing or eliminating a hangover according to one embodiment of the present invention is preferably a hovenia dulcis extract and 100 to 800 parts by weight of alulose, and more preferably 100 to 800 parts by weight of an extract of Hovenia 30 to 50 parts by weight of dulcis extract and 250 to 600 parts by weight of alulose.

According to another embodiment of the present invention, the composition for preventing or eliminating a hangover is selected from the group consisting of red ginseng extract or Hovenia dulcis extract, and a complex saccharide. The composite saccharide comprises 80 to 99.9 wt% of alululose and 0.1 to 20 wt% of fructose, based on the total weight of the complex saccharide, preferably 90 to 98 wt% of allylose and 1 to 10 wt% By weight based on the total weight of the complex sugar, 0.1 to 2% by weight of a glucose oligomer having a degree of polymerization of 2 or more and 92 to 98% by weight of allylose, 1 to 8% by weight of fructose and 2 to 10% by weight of a glucose oligomer having a degree of polymerization of 2 or more. Preferably, the composition for preventing or eliminating a hangover according to the present invention comprises 100 to 800 parts by weight of a complex saccharide based on 100 parts by weight of the red ginseng extract, more preferably 250 to 300 parts by weight of a complex saccharide 250 To 600 parts by weight. In addition, the composition for preventing or eliminating a hangover according to one embodiment of the present invention is preferably a hovenia dulcis extract, and more preferably 400 to 1500 parts by weight of a complex saccharide based on 100 parts by weight of the extract of Hovenia dulcis extract of the present invention, 600 to 1400 parts by weight of a complex saccharide. The composition for preventing or eliminating hangover according to one embodiment of the present invention preferably comprises 20 to 60 parts by weight of a Hovenia dulcis extract and 100 to 800 parts by weight of a complex saccharide based on 100 parts by weight of the red ginseng extract, more preferably heotgae tree (Hovenia based on the 100 parts by weight of ginseng extract 30 to 50 parts by weight of dulcis extract and 250 to 600 parts by weight of complex saccharides.

Hereinafter, the composition for preventing or eliminating a hangover according to the present invention will be described separately for each constituent.

Red ginseng extract or Hinoki ( Hovenia dulcis ) Extract

In the present invention, red ginseng extract or Hovenia dulcis ) extract can be prepared by various methods. As a raw material for obtaining the red ginseng extract of the present invention, red ginseng root, circular red ginseng, red ginseng or other red ginseng can be used. The above-mentioned red ginseng bamboo is manufactured in a state in which the head, body and leg portions are stuck together, and round red ginseng means that the raw ginseng is produced in the original shape of the hair, body, legs and legs. Also, red ginseng refers to a product made from legs or roots separated from manufactured red ginseng, and other ginseng refers to ginseng red ginseng, sliced red ginseng or ground red ginseng. As a raw material for obtaining the Hovenia dulcis Thunb extract of the present invention, various organs or parts such as leaves, stems, roots, fruits and the like of Hovenia wood can be used, and it is preferable to use Hovenia dulcis. On the other hand, in order to obtain an extract from red ginseng or hinoki, an extraction process is required. As the extraction method, a conventional extraction method known in the art can be used, for example, a solvent extraction method. The extraction solvent that can be used when preparing the red ginseng extract or hornbug extract by the solvent extraction method is water, a low alcohol having 1 to 4 carbon atoms (for example, methanol, ethanol, propanol and butanol), or a mixture thereof The solvent may be selected from the group consisting of lower alcohols, propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane, Alcohol, or a mixture thereof. When water is used as the extraction solvent, the water is preferably hot water. When an alcohol is used as an extraction solvent, the alcohol is preferably a lower alcohol having 1 to 4 carbon atoms, and the lower alcohol is more preferably selected from methanol or ethanol. In addition, when the functional alcohol is used as the extraction solvent, the alcohol content is preferably 50 to 90%. It is obvious to those skilled in the art that red ginseng extract or hinoki extract can be obtained not only by using the above-mentioned extraction solvent but also by using other extraction solvent. For example, decompression by carbon dioxide, extraction by supercritical extraction at high temperature, extraction by extraction using ultrasound, separation by ultrafiltration membrane with constant molecular weight cut-off value, various chromatography (size, charge, hydrophobic or affinity The active fraction obtained through various purification and extraction methods, such as separation using the above-described method for separation according to sex, is also included in the extract of the present invention. The supercritical fluid extraction refers to supercritical fluid extraction. Generally, the supercritical fluid is extracted from the liquid and the liquid when the gas reaches the critical point at high temperature and high pressure. (J. Chromatogr. A. 1998; 479: 200-205). In addition, it has been reported that the supercritical fluid has a polarity similar to that of a nonpolar solvent. Carbon dioxide is a supercritical fluid with both liquid and gaseous nature, with the operation of supercritical fluid equipment reaching its critical pressure and temperature, resulting in increased solubility in fat-soluble solutes. When supercritical carbon dioxide passes through an extraction vessel containing a certain amount of sample, the lipophilic substance contained in the sample is extracted into supercritical carbon dioxide. When the supernatant carbon dioxide containing a small amount of cosolvent is passed through the sample remaining in the extraction vessel after extracting the lipid-soluble substance, components that were not extracted by pure supercritical carbon dioxide can be extracted. The supercritical fluid used in the supercritical extraction method of the present invention can effectively extract an active ingredient by using a mixed fluid in which a cosolvent is further mixed with supercritical carbon dioxide or carbon dioxide. These co-solvents may be used alone or in admixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether. Most of the extracted samples contain carbon dioxide. Since the carbon dioxide is volatilized into air at room temperature, the extract obtained by the above method can be used as a cosmetic composition, and the cosolvent can be removed by a reduced pressure evaporator. In addition, the ultrasonic extraction method is an extraction method using energy generated by ultrasonic vibration. Ultrasonic waves can destroy an insoluble solvent contained in a sample in a water-soluble solvent. Due to the high local temperature, Since the kinetic energy of the reactant particles is increased, sufficient energy required for the reaction is obtained. By inducing the high pressure by the shock effect of the ultrasonic energy, the mixing effect of the substance contained in the sample and the solvent is enhanced, thereby increasing the extraction efficiency. As the extraction solvent which can be used for the ultrasonic extraction method, one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether can be used. The extracted sample is recovered by vacuum filtration, and the filtrate is recovered, and the extract is removed by a vacuum evaporator and freeze-dried to obtain an extract.

Alulous  or Alulous  Complexed saccharides containing

In the present invention, alulose is also referred to as D-psicose as an epimer of carbon number 3 of fructose. D-allulose has a 70% sweetness compared to sugar (Oshima 2006), but it is a functional monosaccharide applicable to low-calorie sweeteners (Matsuo et al. 2002) because the calories are about 0.3%. In addition, D-allulose inhibits glucose uptake and regulates blood sugar. In addition, D-allulose inhibits the accumulation of abdominal fat by inhibiting the enzyme activity involved in lipid synthesis in the liver (Matsuo et al., 2001; et al. 2010; Hossain et al. 2011). In addition, various physiological activities of D-allulose have been reported. In the present invention, the allylose-containing complex saccharide means that the allylus content is at least a predetermined amount based on the total weight of the complex saccharide. The allylose-containing complex saccharide may be prepared by mixing alulose and other saccharides at a predetermined ratio, 3-epimerase or D-allyl 3-epimerase to produce fructose by converting the fructose into an alulose and passing the reaction product through chromatography to collect fractions having a predetermined purity of alulose have.

The specific form of the composition for prevention or elimination of hangover

The composition for preventing or eliminating a hangover according to the present invention may be formulated into a pharmaceutical composition, a food additive, a food composition (particularly, a health functional food) depending on the intended use or aspect. In addition, red ginseng extract or Hovenia ( Hovenia dulcis extract and at least one compound selected from alulose or allylose-containing complex saccharides corresponds to an active ingredient for prevention or elimination of hangover, and the content of the active ingredient is also determined by the specific form of the composition, use And may be adjusted in various ranges depending on purposes or aspects.

The content of the active ingredient in the pharmaceutical composition according to the present invention is not particularly limited and is, for example, 0.01 to 99% by weight, preferably 0.5 to 50% by weight, more preferably 1 to 30% by weight, Lt; / RTI > In addition, the pharmaceutical composition according to the present invention may further contain, in addition to the active ingredient, an additive such as a pharmaceutically acceptable carrier, excipient or diluent. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition of the present invention can be formulated into a formulation for oral administration or parenteral administration by a conventional method, and can be formulated into a pharmaceutical composition such as a filler, an extender, a binder, a wetting agent, a disintegrant, Diluents or excipients. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used. Further, it can be suitably formulated according to each disease or ingredient, using appropriate methods in the art or by the method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA. The pharmaceutical composition of the present invention may be administered orally or parenterally to a mammal including a human according to a desired method. Examples of the parenteral administration method include external dermal application, intraperitoneal injection, intramuscular injection, subcutaneous injection, intravenous injection, Intravenous injection or intra-thoracic injection. The dosage of the pharmaceutical composition of the present invention is not limited as long as it is a pharmacologically effective amount and is not limited as long as it depends on the body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, Varies. The typical daily dose of the pharmaceutical composition of the present invention is not particularly limited, but is preferably 0.1 to 3000 mg / kg, more preferably 1 to 2000 mg / kg, based on the active ingredient, once a day Or may be administered in divided doses.

The content of the active ingredient in the food composition according to the present invention is 0.01 to 99% by weight, preferably 0.1 to 50% by weight, more preferably 0.5 to 25% by weight based on the total weight of the composition, but is not limited thereto. The food composition of the present invention may be in the form of a pill, a powder, a granule, an infusion, a tablet, a capsule, or a liquid preparation. Examples of the food include meat, sausage, bread, chocolate, candy, snack, Other noodles, gums, dairy products including ice cream, various soups, drinks, tea, functional water, drinks, alcoholic beverages and vitamin complexes. In addition to the active ingredient, the food composition of the present invention may contain a pharmaceutically acceptable carrier, various flavors or natural carbohydrates as an additional ingredient. In addition, the food composition of the present invention can be used as a food composition containing various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, , A carbonating agent used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The above-mentioned natural carbohydrates are sugar alcohols such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Natural flavors such as tau Martin and stevia extract, and synthetic flavors such as saccharin and aspartame may be used as the flavor.

Hereinafter, the present invention will be described more specifically by way of examples. However, the following examples are intended to clearly illustrate the technical features of the present invention and do not limit the scope of protection of the present invention.

1. Preparation of a recombinant strain producing D-alulose 3-epimerase

Genomic DNA was extracted from Flavonifractor plautii KCTC 5970, which was distributed at the Korea Microorganism Resource Center, and used as a template, and a gene encoding D-allose 3-epimerase Polymerase chain reaction (PCR) was carried out using a primer for cloning the polynucleotide of SEQ ID NO: 2 and Ex-Taq (TAKARA) polymerase. Table 1 below shows Flavonifractor < RTI ID = 0.0 > alulose 3-epimerase from the genomic DNA of E. coli , E. coli , E. coli , C. plautii , and the like. The primers shown in Table 1 were manufactured by Bioneer co., KR.

SEQ ID NO: Type of primer The base sequence (5 '- > 3') The restriction enzyme recognition site contained in the primer 3 Forward primer for cloning gene of alulose epimerase CGG CAT ATG AAC CCG ATT GGA ATG CAC TAC Nde I 4 Reverse primer for cloning gene of alulos epimerase CGG CTC GAG TTA CGC GGT CAG CTC CTT GAG G Xho I

Then, the target DNA was isolated from the PCR product using the gel extraction kit (Qiagen), bound to Easy T-vector (promega), and the base sequence analysis of the isolated DNA was performed by Bioneer co., KR Respectively. As a result, it was confirmed that the target DNA amplified by PCR corresponds to the polynucleotide of SEQ ID NO: 2. The recombinant expression vector pET-FDPE was constructed by inserting the target DNA amplified by the PCR reaction into the same restriction enzyme recognition site of the pET15b vector (Novagen), which is an expression vector, using restriction enzymes Nde I and Xho I. Then, E. coli BL21 (manufactured by RBC, Taipei, Taiwan), a competent cell, was transformed with recombinant expression vector pET-FDPE using electroporation to prepare a recombinant strain.

A single colony of the transformed recombinant strain was inoculated in 15 ml of LB-ampicilline medium (Difco) and pre-cultured at 37 DEG C and 200 rpm for about 6 hours. Then, the preculture was inoculated into 500 ml of LB-ampicilline medium and cultured under shaking conditions at 37 DEG C and 200 rpm. Then, when the absorbance (at 600 nm) of the culture was 0.5, IPTG was added at a concentration of 0.1 mM to induce the overexpression of the target enzyme. At the induction of overexpression, the culture was switched to 16 ° C and 150 rpm and maintained for about 16 hours. Then, the culture of the recombinant strain was centrifuged at 13000 rpm for 2 minutes to remove the supernatant, and the cells of the recombinant strain were recovered.

Cells of the recovered recombinant strains were suspended in lysis buffer (50 mM Tris-HCl 300 mM NaCl pH 8.0, 10 mM imidazol) and sonicated to disrupt the cells. The cell lysate was centrifuged at 13000 rpm for 10 minutes to collect only the supernatant. The supernatant was collected and applied to a Ni-NTA column (Bio-Rad, Profinia) previously equilibrated with lysis buffer, and then eluted with 50 mM Tris-HCl 300 mM NaCl pH 8.0 in 20 mM imidazole Buffer solution containing 200 mM imidazole was sequentially flowed. Finally, the target protein was eluted by flowing 50 mM Tris-HCl 300 mM NaCl pH 8.0 and 200 mM imidazol. The eluted protein was found to be a D-allylose 3-epimerase consisting of the amino acid sequence of SEQ ID NO: 1.

The applicant of the present invention has filed a patent application and a patent application for a method for producing alulose using the D-alulose 3-epimerase, the recombinant strain producing the D-alulose 3-epimerase, and the D-alulose 3-epimerase or the recombinant strain Patent registration (Korean Patent Registration No. 10-1473918). The present application includes a method for producing aluloses disclosed in Korean Patent Registration No. 10-1473918.

2. Alulous  Preparation of Syrup and Analysis of Composition of Sugars

(1) Preparation of alulose-containing syrup

Production Example 1

(Sugar composition: 80 weight% of fructose, 16 weight% of glucose and 4 weight% of glucose oligomer having a polymerization degree of 2 or more) having a total solid concentration of 35 Brix was placed in a 5 L reaction tube, The pH of the fructose-containing solution was adjusted to a range of about 7-8 using sodium hydroxide solution. Thereafter, the temperature of the fructose-containing solution was maintained at 60 DEG C by using a constant temperature water bath, and the cells of the recombinant strain developed by the applicant of the present invention were added at a level of 1.5% by weight based on the total solid content of the fructose- For a period of time to convert fructose to alulose. Thereafter, the pH of the cell reaction liquid was adjusted to 3 and maintained for 1 hour to terminate the cell reaction. Thereafter, the bacterial reaction solution was filtered with a diatomaceous earth filter using a filter press apparatus (incarnation machine) to remove impurities. Thereafter, the filtrate was decolorized with activated carbon, and purified using an ion exchange resin. Specifically, the decolorized filtrate was mixed with a cation exchange resin (SCR-B, Samyang Corp.), an anion exchange resin (S4268, Lanxess) and a mixed ion exchange resin (cation exchange resin (SCR- ) Were mixed in a weight ratio of 1: 2] in order to conduct ion purification. Thereafter, the purified liquid was concentrated to a solid concentration of about 75 Brix using a vacuum concentrator, and the concentrate was used as an alulose-containing syrup in a later-described experiment.

Production Example 2

(Sugar composition: 80 weight% of fructose, 16 weight% of glucose and 4 weight% of glucose oligomer having a polymerization degree of 2 or more) having a total solid concentration of 35 Brix was placed in a 5 L reaction tube, The pH of the fructose-containing solution was adjusted to a range of about 7-8 using sodium hydroxide solution. Thereafter, the temperature of the fructose-containing solution was maintained at 60 캜 by using a constant temperature water bath, and the cells of the recombinant strain developed by the applicant of the present invention were added at a level of 1.5% by weight based on the total solid content of the fructose- For a period of time to convert fructose to alulose. Thereafter, the pH of the cell reaction liquid was adjusted to 3 and maintained for 1 hour to terminate the cell reaction. Thereafter, the bacterial reaction solution was filtered with a diatomaceous earth filter using a filter press apparatus (incarnation machine) to remove impurities. Thereafter, the filtrate was subjected to primary decolorization using activated carbon, and then primary purification was performed using an ion exchange resin. Specifically, the decolorized filtrate was mixed with cation exchange resin (SCR-B, Samyang Corp.), anion exchange resin (S4268, Lanxess) and mixed ion exchange resin (cation exchange resin (SCR- ) Were mixed in a weight ratio of 1: 2] in order to carry out primary ion purification. Thereafter, the primary purification solution was firstly concentrated using a vacuum concentrator to a solid concentration of about 60 Brix, and the primary concentrate was passed through a calcium-type fixed bed ion exchange chromatography to obtain an allylose purity of about 95% Separate liquid was obtained. Thereafter, the separated liquid was subjected to second decolorization using activated carbon, and then passed through a cation-exchange resin and an anion-exchange resin in order to perform secondary purification. Thereafter, the second purified liquid was secondarily concentrated to a solid concentration of about 75 Brix using a vacuum concentrator, and the second concentrated liquid was used as an allylose-containing syrup in a later-described experiment.

(2) Analysis of sugar composition ratio of alulose-containing syrup

The saccharide compositions of the alulose-containing syrups prepared in Preparation Example 1 and Preparation Example 2 were analyzed using high performance liquid chromatography (HPLC). As a result, the saccharide composition of the alulose-containing syrup prepared in Preparation Example 1 was 56% by weight of fructose, 16% by weight of glucose, 4% by weight of allylose and 4% by weight of glucose oligomer having a degree of polymerization of 2 or more, The saccharide composition of the los containing syrup was 95% by weight of allylose, 4% by weight of fructose and 1% by weight of glucose oligomer having a degree of polymerization of 2 or more. The saccharide content was based on the total weight of the saccharide solids contained in the alululose-containing syrup and was calculated based on the peak area of the HPLC analysis results. The HPLC analysis conditions are as follows.

* Column type: Aminex HPX-87C carbohydrate column

* Column temperature: 80 ° C

* Sample volume (inject volume): 10 μl

* Sample flow rate: 0.6 ml / min

* Run time: 25 min

3. For various samples Aldehyde  Dehydrogenase ( aldehyde dehydrogenase , ALDH ) Active measurement

(1) Preparation of samples for measurement of ALDH activity

The solid concentration of each of the hull nut extract, the red ginseng extract, the alulose-containing syrup prepared in Preparation Example 1, the alulose-containing syrup prepared in Preparation Example 2, the white sugar, the high fructose syrup, (Brix) to prepare a sample for ALDH activity measurement. The Hovenous Fruit Extract was prepared by adding purified water having a weight of about 10 times to dried Hovenia dulcis powder and hot-extraction at about 80 ° C for about 10 hours, filtering it using a filter paper, and adjusting the final solid concentration of the filtrate to 2 Brix ). In addition, the red ginseng extract was prepared by adding purified water to red ginseng root and hot-extracted at about 85 ° C for about 72 hours, filtered using a filter paper, and the final solid concentration of the filtrate was adjusted to 2 brix. The whiteness is a product whose moisture content is close to 0% by weight. The high fructose syrup has a solids concentration of about 75 Brix and the sugar composition is about 55 weight percent glucose, about 40 weight percent glucose, and about 5 weight percent glucose oligomer having a degree of polymerization of at least 2, based on the total weight of sugar solids. The hydrolyzed glucose and fructose are products having a water content of about 5% by weight.

(2) Methods and results of measurement of ALDH activity

ALDH activity is an enzyme that produces acetate in acetaldehyde, and has been used to generate NADH from NAD (Koivula and Koivusalo, 1975). 0.1 mL of distilled water, 0.1 mL of 1.0 M Tris-HCl buffer (pH 8.0), 0.3 mL, 20 mM NAD + 0.1 mL, 0.1 M acetaldehyde, 0.1 mL of 3.0 M KCl, 0.1 mL of 0.33 M 2-mercaptoethanol and 0.1 mL of the sample were mixed After incubation at 25 ° C for 10 minutes, 0.1 mL of ALDH (1 unit / ml) was added, and the reaction solution was adjusted to 3 mL. The mixture was placed in a cuvette and preliminarily cultured at 25 ° C for 5 minutes. Change was measured. At this time, distilled water was used as a control instead of the sample. ALDH activity was expressed as the ratio of the maximum absorbance at the end of the reaction to the maximum absorbance of the control, and was calculated by the following equation.

ALDH activity (%) = (B / A) x 100

A: Maximum absorbance of the control

B: Maximum absorbance of the experimental group

The following Table 2 shows the results of the analysis of the solid content concentration of the hull fruit extract, the red ginseng extract, the alulose-containing syrup prepared in Preparation Example 1, the alulose-containing syrup prepared in Preparation Example 2, Were adjusted so as to be 2 brix, and their ALDH activity was measured.

sample ALDH activity (%) Houttuynia cordata extract 142.86 Red ginseng extract 139.29 The alulose-containing syrup of Preparation Example 1 46.43 The alulose-containing syrup of Preparation Example 2 85.71 Jungbang 3.57 High fructose syrup 32.14 Functional Crystalline Glucose 17.86 Crystalline fructose 17.86

As shown in Table 2, a diluted solution (aliquot concentration: 2 Brix) of the alululose-containing syrup prepared in Preparation Example 1, the alululose-containing syrup prepared in Preparation Example 2, whitening sugar, high fructose syrup, Showed that ALDH activity was lower than that of control, distilled water.

4. Preparation of a composition for relieving hangovers Aldehyde  Dehydrogenase ( aldehyde  dehydrogenase, ALDH ) Active measurement

(1) Preparation of composition for removing hangover

Production Example 3

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 95 parts by weight of the mixture of Hovenia dulcus extract / red ginseng extract and 5 parts by weight of alulose-containing syrup (solid concentration: 75 Brix) prepared in Preparation Example 1 were mixed to prepare a composition for removing hangover.

Production Example 4

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 93 parts by weight of the mixture of Hovenous fruit extract / red ginseng extract and 7 parts by weight of alulose-containing syrup (solid concentration: 75 Brix) prepared in Preparation Example 1 were mixed to prepare a composition for removing hangover.

Production Example 5

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 90 parts by weight of the mixture of Hovenia dulcus extract / red ginseng extract and 10 parts by weight of alulose-containing syrup (solid concentration: 75 Brix) prepared in Preparation Example 1 were mixed to prepare a composition for removing hangover.

Production Example 6

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Thereafter, 95 parts by weight of the mixture of Houttuynia fruit extract / red ginseng extract and 5 parts by weight of alulose-containing syrup (solid concentration: 75 Brix) prepared in Preparation Example 2 were mixed to prepare a composition for removing hangover.

Production Example 7

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Subsequently, 93 parts by weight of a mixture of Hovenia dulcus extract / red ginseng extract and 7 parts by weight of alulose-containing syrup (solid concentration: 75 Brix) prepared in Preparation Example 2 were mixed to prepare a composition for removing hangover.

Production Example 8

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Thereafter, 90 parts by weight of the mixture of Houttuynia fruit extract / red ginseng extract and 10 parts by weight of alulose-containing syrup (solid concentration: 75 Brix) prepared in Preparation Example 2 were mixed to prepare a composition for removing hangover.

Production Example 9

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 95 parts by weight of a mixture of Hovenia dulcus extract / red ginseng extract and 5 parts by weight of water-soluble glucose (moisture content: 5% by weight) were mixed to prepare a composition for removing hangover.

Production Example 10.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 93 parts by weight of a mixture of Houttuynia cordata extract / red ginseng extract and 7 parts by weight of water-soluble glucose (moisture content: 5% by weight) were mixed to prepare a composition for removing hangover.

Production Example 11.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Thereafter, 90 parts by weight of a mixture of Hovenia dulcus extract / red ginseng extract and 10 parts by weight of water-soluble glucose (moisture content: 5% by weight) were mixed to prepare a composition for removing hangover.

Production Example 12.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Thereafter, 95 parts by weight of a mixture of Hovenia dulcus extract / red ginseng extract and high fructose syrup (solids concentration: 75 Brix; glucose composition: about 55% by weight of glucose, about 40% by weight of glucose, And 5 parts by weight of an oligomer) were mixed to prepare a composition for removing hangover.

Production Example 13.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Thereafter, 93 parts by weight of a mixture of Houttuynia cordata extract / red ginseng extract and a high fructose syrup (solids concentration: 75 Brix; glucose composition: about 55% by weight of fructose, about 40% by weight glucose, And 5 parts by weight of an oligomer) were mixed to prepare a composition for removing hangover.

Production Example 14.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 90 parts by weight of a mixture of Hovenia dulcus extract / red ginseng extract and high fructose syrup (solids concentration: 75 Brix; glucose composition: about 55% by weight of glucose, about 40% by weight of glucose, And 5 parts by weight of an oligomer) were mixed to prepare a composition for removing hangover.

Production Example 15.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Thereafter, 95 parts by weight of a mixture of Houttuynia cordata extract / red ginseng extract and 5 parts by weight of crystalline fructose (moisture content: about 5% by weight) were mixed to prepare a composition for removing hangover.

Production Example 16.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Thereafter, 93 parts by weight of a mixture of Houttuynia cordata extract / red ginseng extract and 7 parts by weight of crystalline fructose (moisture content: about 5% by weight) were mixed to prepare a composition for removing hangover.

Production Example 17.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 90 parts by weight of the mixture of Houttuynia cordata extract / red ginseng extract and 10 parts by weight of crystalline fructose (moisture content: about 5% by weight) were mixed to prepare a composition for removing hangover.

Production Example 18.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Thereafter, 95 parts by weight of a mixture of Houttuynia cordata extract / red ginseng extract and 5 parts by weight of a whitish white (moisture content: about 0% by weight) were mixed to prepare a composition for removing hangover.

Production Example 19.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 93 parts by weight of a mixture of Houttuynia fruit extract / red ginseng extract and 7 parts by weight of a whitish white (moisture content: about 0% by weight) were mixed to prepare a composition for removing hangover.

Production Example 20.

Hovenia dulcis extract / red ginseng extract mixture was prepared by mixing Houttuynia fruit extract (solid concentration: 2 Brix) and red ginseng extract (solid concentration 2 Brix) at a weight ratio of 3: 7. Then, 90 parts by weight of a mixture of Houttuynia cordata extract / red ginseng extract and 10 parts by weight of a whitish white (moisture content: about 0 wt.%) Were mixed to prepare a composition for removing hangovers.

(2) Production of ALDH activity measuring sample and measurement of ALDH activity

The composition for dissolving hangover prepared in Preparative Examples 3 to 20 was diluted with purified water to a solid concentration of 2 brix to prepare a sample for ALDH activity measurement. ALDH activity is an enzyme that produces acetate in acetaldehyde, and has been used to generate NADH from NAD (Koivula and Koivusalo, 1975). 0.1 mL of distilled water, 0.1 mL of 1.0 M Tris-HCl buffer (pH 8.0), 0.3 mL, 20 mM NAD + 0.1 mL, 0.1 M acetaldehyde, 0.1 mL of 3.0 M KCl, 0.1 mL of 0.33 M 2-mercaptoethanol and 0.1 mL of the sample were mixed After incubation at 25 ° C for 10 minutes, 0.1 mL of ALDH (1 unit / ml) was added, and the reaction solution was adjusted to 3 mL. The mixture was placed in a cuvette and preliminarily cultured at 25 ° C for 5 minutes. Change was measured. At this time, distilled water was used as a control instead of the sample. ALDH activity was expressed as the ratio of the maximum absorbance at the end of the reaction to the maximum absorbance of the control, and was calculated by the following equation.

ALDH activity (%) = (B / A) x 100

A: Maximum absorbance of the control

B: Maximum absorbance of the experimental group

Table 3 below shows the results of measuring the ALDH activity of the compositions for removing hangover prepared in Preparative Examples 3 to 20 so as to have a solid concentration of 2 brix.

sample ALDH activity (%) sample ALDH activity (%) Production Example 3 120.19 Production Example 12 85.11 Production Example 4 81.35 Production Example 13 80.82 Production Example 5 71.99 Production Example 14 82.41 Production Example 6 142.86 Production Example 15 69.10 Production Example 7 150.0 Production Example 16 47.60 Production Example 8 157.14 Production Example 17 53.21 Production Example 9 76.15 Production Example 18 73.04 Production Example 10 43.10 Production Example 19 63.24 Production Example 11 46.12 Production example 20 46.92

As shown in Table 3, in the case of high-fructose syrup, hydrolyzed glucose, crystalline fructose and alulose-containing syrup having low purity, it can be mixed with Hovenia dulcis extract or red ginseng extract to impart sweetness characteristics. However, Respectively. On the other hand, in the case of alulose-containing syrup having high purity of alulose, it was mixed with Hovenia dulcis extract or red ginseng extract to impart sweetness characteristics and increase ALDH activity. From the above results, it can be seen that alulose or high-purity alulose-containing syrup acts as a booster to increase the ALDH activity of Hornberry fruit extract or red ginseng extract.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Therefore, the scope of the present invention should be construed as including all embodiments falling within the scope of the appended claims.

<110> DAESANG CORPORATION <120> Composition for preventing or relieving hangover including          allulose <130> DP-16-946 <160> 4 <170> Kopatentin 2.0 <210> 1 <211> 294 <212> PRT <213> Artificial Sequence <220> &Lt; 223 > D-allulose 3-epimerase derived from Flavonifractor plautii <400> 1 Met Asn Pro Ile Gly Met His Tyr Gly Phe Trp Ser His Asn Trp Asp   1 5 10 15 Glu Ile Ala Tyr Ile Pro Leu Met Glu Lys Leu Ala Trp Leu Gly Phe              20 25 30 Asp Ile Cys Glu Val Ala Ser Ala Glu Trp Gly Tyr Tyr Asp Asp Ala          35 40 45 Arg Leu Arg Glu Leu Lys Ala Cys Ala Asp His Asn Gly Leu Gly Ile      50 55 60 Thr Tyr Ser Ile Gly Leu Glu Ala Lys Tyr Asp Leu Ala Ser Asp Asp  65 70 75 80 Pro Ala Val Arg Glu Asn Gly Ile Arg His Val Thr Arg Ile Leu Glu                  85 90 95 Ser Met Pro Lys Val Gly Ala Ile Leu Asn Gly Val Ser Tyr Ala             100 105 110 Gly Trp Gln Ala Leu Pro Asp His Gly Ile Thr Leu Asp Glu Lys Arg         115 120 125 Arg Lys Glu Glu Leu Ala Leu Glu Ser Met Ser Arg Leu Met Lys Val     130 135 140 Ala Glu Asp Cys Gly Val Leu Tyr Cys Cys Glu Val Val Asn Arg Phe 145 150 155 160 Glu Gln Tyr Leu Leu Asn Thr Ala Lys Glu Gly Val Glu Phe Val Lys                 165 170 175 Arg Leu Gly Ser Pro Asn Ala Arg Val Leu Leu Asp Thr Phe His Met             180 185 190 Asn Ile Glu Glu Asp Ser Met Val Asp Ala Ile Leu Glu Ala Gly Pro         195 200 205 Trp Leu Gly His Phe His Val Gly Glu Asn Asn Arg Arg Pro Ala Gly     210 215 220 Ser Thr Asn Arg Leu Pro Trp Lys Asp Met Ala Ala Ala Leu Lys Gln 225 230 235 240 Val Asn Tyr Gln Gly Ala Ile Val Met Glu Pro Phe Val Leu Met Gly                 245 250 255 Gly Thr Ile Pro Tyr Asp Ile Lys Val Trp Arg Asp Leu Ser Gly Gly             260 265 270 Ala Gly Glu Ala Gly Leu Asp Glu Met Ala Gly Arg Ala Cys Arg Phe         275 280 285 Leu Lys Glu Leu Thr Ala     290 <210> 2 <211> 885 <212> DNA <213> Artificial Sequence <220> <223> polynucleotide coding D-allulose 3-epimerase derived from          Flavonifractor plautii <400> 2 atgaacccga ttggaatgca ctacggcttc tggagccaca actgggacga gattgcatac 60 atacccctga tggagaagct ggcctggctg ggctttgaca tctgcgaggt ggcctccgcc 120 gagtggggct attacgacga cgccaggctg cgggagctga aggcctgcgc cgatcacaac 180 ggcctgggca tcacctattc catcggcctg gaggccaaat acgacctggc cagcgacgat 240 ccggcggtgc gggagaacgg catccgccat gtcacccgca tcctggagag catgcccaag 300 gtgggggcgg ccatcctcaa cggcgtgtcc tacgccgggt ggcaggccct gcccgaccac 360 ggaatcaccc tggacgagaa gcgccgcaag gaggagcttg ccctggagtc catgtcccgg 420 ctcatgaagg tggcggagga ctgcggcgtg ctctactgct gcgaggtggt caaccgcttc 480 gagcagtacc tgctcaacac cgccaaagag ggcgtggagt ttgtcaagcg cctgggcagt 540 cccaacgccc gggtgctgct ggataccttc cacatgaaca tcgaggagga cagcatggtg 600 gacgccattc tggaggcggg cccctggctg gggcatttcc acgtggggga gaacaaccgc 660 cgccccgccg gctccaccaa ccgcctgccc tggaaggaca tggccgccgc cctcaagcag 720 gtgaactacc agggggccat tgtgatggag cccttcgtgc tcatgggggg taccattccc 780 tatgatatca aggtctggcg ggatctcagc ggcggggccg gggaggccgg gctggacgag 840 atggcgggcc gggcctgccg gttcctcaag gagctgaccg cgtaa 885 <210> 3 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> forward primer for cloning D-allulose 3-epimerase gene <400> 3 cggcatatga acccgattgg aatgcactac 30 <210> 4 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for cloning D-allulose 3-epimerase gene <400> 4 cggctcgagt tacgcggtca gctccttgag g 31

Claims (8)

20 to 60 parts by weight of Hovenia dulcis extract and 100 to 800 parts by weight of saccharides based on 100 parts by weight of red ginseng extract,
Wherein the saccharide is made of alulose alone or comprises 95 to 99.9% by weight of alulose and 0.1 to 5.0% by weight of fructose based on the total weight of saccharides.
delete delete The composition for prevention or elimination of hangover according to claim 1, which comprises 30 to 50 parts by weight of Hovenia dulcis extract and 250 to 600 parts by weight of saccharides based on 100 parts by weight of the red ginseng extract.
The method according to claim 1, wherein the saccharide comprises 95 to 98% by weight of allylose, 1 to 4% by weight of fructose, and 0.1 to 2% by weight of glucose oligomer having a degree of polymerization of 2 or more, / RTI &gt;
delete delete 2. The method according to claim 1, wherein the saccharide comprises a fructose-containing solution having a saccharide composition of 80 wt% glucose, 16 wt% glucose and 2 or more glucose oligomers having a polymerization degree of 2 or more, And the mixture was reacted at 60 DEG C for 30 hours to convert fructose to alululose. The reaction solution was filtered, and the filtrate was decolorized. The decolored filtrate was applied to a cation exchange resin, an anion exchange resin, And then passing the ion-purified liquid through a calcium-type fixed bed ion exchange chromatography to obtain an allylose-containing syrup-containing saccharide,
Wherein said recombinant strain is a strain transformed by the introduction of a recombinant expression vector expressing D-allose 3-epimerase comprising the amino acid sequence of SEQ ID NO: 1.