WO2011145518A1 - アルデヒド除去組成物 - Google Patents
アルデヒド除去組成物 Download PDFInfo
- Publication number
- WO2011145518A1 WO2011145518A1 PCT/JP2011/061048 JP2011061048W WO2011145518A1 WO 2011145518 A1 WO2011145518 A1 WO 2011145518A1 JP 2011061048 W JP2011061048 W JP 2011061048W WO 2011145518 A1 WO2011145518 A1 WO 2011145518A1
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- WO
- WIPO (PCT)
- Prior art keywords
- acetaldehyde
- aldehyde
- gluconobacter
- composition according
- cells
- Prior art date
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- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
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- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- 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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/02—Antidotes
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- 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
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- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/16—Tea extraction; Tea extracts; Treating tea extract; Making instant tea
- A23F3/166—Addition of, or treatment with, enzymes or microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Definitions
- the present invention relates to a composition for removing aldehyde, for example.
- Non-Patent Document 1 Alcohol (ethanol) is carcinogenic to animals based on the work committee of IARC (the International Agency for Research). We conclude that there is sufficient evidence.
- alcohol After drinking, alcohol is generally decomposed into acetaldehyde by alcohol dehydrogenase in the liver, further oxidized by aldehyde dehydrogenase (aldehyde dehydrogenase) to become acetic acid, and finally decomposed into carbon dioxide and water.
- aldehyde dehydrogenase aldehyde dehydrogenase
- the acetaldehyde produced as an intermediate during the catabolism of ethanol is not only a causative substance of so-called “drunk” symptoms such as flushing of the face, headache, sleepiness, nausea and vomiting, but also a carcinogenic substance of Group I in IARC ( Acetaldehyde derived from alcohol by metabolism is considered to play a major role in the carcinogenicity of alcohol.
- acetaldehyde is a cause of blue odor, off-flavor, and altered odor in brewed beer such as beer and sake, beer-like alcoholic beverages, and distilled spirits such as shochu, and is called a flavor-inhibiting substance (off-flavor).
- off-flavor a flavor-inhibiting substance
- excess acetaldehyde produced by drinking can accumulate in saliva and cause a bad breath ("ripe odor").
- the Japanese population has a high proportion of aldehyde dehydrogenase isozyme type 2 (ALDH2) deficiency responsible for detoxification of acetaldehyde (hetero-deficient ALDH2 * 1 / * 2: about 40% of the total population; homo-deficient ALDH2 * 2 / * 2: Approximately 10%). Since people with this genotype have a slow metabolism rate of acetaldehyde, they become a flusher type whose face becomes remarkably red by drinking (Non-patent Document 2). The flasher type person has about 50 times higher risk of carcinogenesis in the upper digestive organ group (oral cavity, pharynx and esophagus) due to drinking than the non-flasher type (Non-patent Document 3).
- acetaldehyde is secreted into the blood and released by exhalation, and accumulates in saliva.
- Alcohol in the oral cavity is metabolized into acetaldehyde by oral bacteria and accumulates in the upper digestive organs.
- saliva of flasher-type humans with slow acetaldehyde metabolism it has been observed that the acetaldehyde concentration rises significantly after drinking and circulates and stays in the upper digestive tract over several hours to several tens of hours. It has been found that the circulation and retention of acetaldehyde in the upper gastrointestinal tract due to alcohol drinking is the main cause of the increased risk of flasher upper gastrointestinal cancer.
- Alcohol beverages, particularly some distilled liquors originally contain trace amounts of acetaldehyde.
- Non-patent Document 4 As a means for removing acetaldehyde in the oral cavity after drinking, use of L-cysteine (sulfur-containing amino acid), barley green juice, etc. is considered (Non-patent Document 4). This method was devised based on the fact that L-cysteine and flavonoids in green juice add to acetaldehyde to inactivate acetaldehyde. However, the complex formation reaction between flavonoids in L-cysteine or green juice and acetaldehyde is a reversible reaction, and this method has a drawback that acetaldehyde may be regenerated. L-cysteine and green juice itself are substances having a unique taste and odor and are practically difficult to use by taking.
- Patent Document 1 discloses an alcohol-decomposable composition containing acetic acid bacteria that degrade alcohol at pH 3 or lower.
- Patent Document 1 discloses that when an acetic acid bacterium exhibiting alcohol oxidase activity even at pH 3 or lower is ingested before or before drinking alcohol, alcohol is oxidized into acetic acid through acetaldehyde in the stomach by the acetic acid bacterium. Indicate that it is not absorbed in the digestive tract.
- microorganisms In practice, however, such microorganisms also have the ability to oxidize ethanol in most cases, and this method has the disadvantage that a large amount of acetaldehyde is generated from alcohol remaining in the oral cavity by taking rather than before taking it. . Further, it should be noted that components in food such as beer and saliva components significantly increase acetaldehyde generation activity. Therefore, the adverse effect that acetaldehyde increases rather by taking a microorganism having aldehyde-degrading activity is brought about. Such a problem can be solved by purifying acetaldehyde-generating activity from microorganisms or selectively inactivating acetaldehyde-generating activity by applying an appropriate treatment. However, the use of microorganisms having aldehyde-degrading activity still has practical disadvantages such as the generation of harmful substances in the acetaldehyde-degrading enzyme reaction and the necessity of adding a coenzyme.
- Aldehyde oxidase and aldehyde dehydrogenase can be used as the main acetaldehyde-degrading enzymes of microorganisms.
- Aldehyde oxidase uses oxygen molecules to oxidize aldehydes and catalyze a reaction that produces carboxylic acid and hydrogen peroxide. Hydrogen peroxide, one of the reaction products, is a carcinogenic harmful substance, and its production is not preferable.
- aldehyde dehydrogenase catalyzes the oxidation of aldehyde using NAD + , NADP +, etc. as coenzymes. These coenzymes are very expensive. In practice, it is necessary to add a large excess of coenzyme compared to the substrate or to incorporate an oxidized coenzyme regeneration system, but this is not practical from the viewpoint of cost.
- an object of the present invention is to provide a method for removing acetaldehyde accumulated in the oral cavity after drinking from the oral cavity by a safe means.
- the present invention is a carcinogenic risk factor for upper gastrointestinal tract cancer in a flasher, decomposes acetaldehyde accumulated in the oral cavity after drinking, and rapidly reduces the oral concentration to a carcinogenic concentration (2.2 ppm) or less. It is an object of the present invention to provide a method.
- a method of decomposing and removing acetaldehyde using bacteria that are naturally present in the human body, such as food microorganisms and enteric bacteria, or microorganisms that are harmless to humans in the environment is considered. It is done.
- the problem to be satisfied by this method is i) special treatment (purification, heat treatment, etc.) for the purpose of preventing the production of acetaldehyde from alcohol remaining in the oral cavity after drinking or eating or drinking, or preventing acetaldehyde Ii) not requiring the addition of oxidized coenzymes (NAD, etc.) to promote the decomposition and removal of acetaldehyde, and iii) not producing harmful substances in the process of decomposing acetaldehyde, or It is not necessary to add an additive for the purpose of decomposing harmful substances generated in the process of acetaldehyde decomposition.
- special treatment purification, heat treatment, etc.
- the concentration of alcohol ranges from several hundred to several thousand times that of acetaldehyde, and it is necessary to efficiently decompose acetaldehyde under such conditions.
- microorganisms with acetaldehyde-degrading ability have a strong ethanol oxidation activity, and thus acetaldehyde is produced in the presence of a large excess of ethanol. For this reason, there is no known microbial preparation capable of removing acetaldehyde accumulated in the oral cavity after drinking from the oral cavity.
- the present invention includes the following.
- An aldehyde-removing composition containing a microorganism belonging to the genus Gluconobacter having aldehyde-degrading activity, or an extract thereof or a crushed cell, wherein the microorganism belonging to the genus Gluconobacter is more aldehyde than aldehyde-forming activity.
- the said aldehyde removal composition characterized by having high decomposition activity.
- the aldehyde removal composition according to (1) which is a composition for reducing the risk of upper gastrointestinal cancer.
- acetaldehyde which increases the risk of carcinogenesis of the upper gastrointestinal tract associated with drinking habits, and aldehyde which is the cause of odor, and to fundamentally remove the carcinogenic risk factor of the upper gastrointestinal tract and the cause of the odor.
- a possible composition is provided.
- the aldehyde-removing composition according to the present invention contains a microorganism belonging to the genus Gluconobacter having aldehyde-degrading activity, an extract thereof, or a crushed cell. .
- the microorganism belonging to the genus Gluconobacter has higher aldehyde decomposing activity than aldehyde producing activity, and can decompose aldehyde under conditions where the alcohol concentration is higher than the aldehyde concentration.
- the composition according to the present invention can be used for decomposing and removing acetaldehyde that increases the risk of carcinogenesis in the upper gastrointestinal tract associated with drinking habits and the aldehyde that is the cause of odor, and the composition for reducing the risk of suffering from upper gastrointestinal cancer or the composition for removing odor Can be used as a thing.
- examples of upper gastrointestinal cancer include oral cancer, pharyngeal cancer, and esophageal cancer.
- acetaldehyde associated with drinking was classified as Group 1 in 2010, and it is recognized that it is definitely carcinogenic to humans.
- the carcinogenic concentration of acetaldehyde associated with alcohol consumption is estimated to be 2 ppm (50 microM). Therefore, if the acetaldehyde in the oral cavity can be immediately reduced to 2 ppm or less by the aldehyde removing composition according to the present invention, it can be considered that the composition according to the present invention can reduce the risk of carcinogenesis.
- acetaldehyde in the oral cavity after drinking can be decomposed and removed by taking, and acetaldehyde that increases the risk of carcinogenesis in the upper gastrointestinal tract associated with drinking habits can be fundamentally removed.
- examples of the odor include a body odor such as a ripe odor caused by acetaldehyde, an aging odor caused by nonenal, and a blue odor caused by hexanal.
- a body odor such as a ripe odor caused by acetaldehyde, an aging odor caused by nonenal, and a blue odor caused by hexanal.
- acetaldehyde which is a mature odor substance in the oral cavity after drinking, can be decomposed and removed by taking, and the cause of odor can be fundamentally removed.
- the composition according to the present invention can also be used for removing body odor.
- the composition which concerns on this invention can also be used for the blue odor removal in food-drinks, such as soy milk.
- examples of the aldehyde compound include acetaldehyde, nonenal, hexanal and the like.
- the microorganism used in the present invention has higher aldehyde decomposition activity than aldehyde (for example, acetaldehyde) production activity, and aldehydes under conditions where alcohol (for example, ethanol) concentration is higher than aldehyde (for example, acetaldehyde) concentration. It can be disassembled.
- the microorganism used in the present invention is efficient in the presence of an alcohol such as ethanol in a large excess (for example, several hundred to several thousand times in molar ratio) with respect to the aldehyde such as acetaldehyde. Can be decomposed.
- Gluconobacter that does not produce harmful substances such as hydrogen peroxide during the decomposition reaction.
- microorganisms belonging to the genus Gluconobacter include microorganisms belonging to species such as Gluconobacter kondonii, Gluconobacter kanchanaburiensis.
- strains that can be used in the present invention include Gluconobacter kondonii RC NBRC3266 and Gluconobacter chanchanaburiensis NBRC103587. These strains can be obtained from the Biological Genetic Resource Department (NBRC) (2-5-8 Kazusa Kamashi, Kisarazu City, Chiba Prefecture, Japan) of the National Institute for Product Evaluation and Technology (NITE).
- NBRC Bio Genetic Resource Department
- the microorganism used in the present invention is a bacterium belonging to the genus Gluconobacter, which does not produce acetaldehyde in the presence of ethanol and can efficiently decompose acetaldehyde, or acetaldehyde from ethanol even in a liquor / saliva mixed system.
- Any microorganism may be used as long as it is a microorganism that decomposes acetaldehyde without producing it and does not produce harmful substances such as hydrogen peroxide during the decomposition reaction.
- ethanol does not produce acetaldehyde in the presence of ethanol
- ethanol is not oxidized at all, or even if it has weak ethanol oxidation activity (ie, acetaldehyde production activity), it has acetaldehyde oxidation activity (ie, acetaldehyde). It is extremely weak compared to (degradation activity).
- the ethanol oxidation activity is about 1/10, preferably about 1/50 of the acetaldehyde oxidation activity.
- mutant strains can be used without mutation treatment with ethyl methanesulfonic acid, a commonly used mutation agent, treatment with other chemical substances such as nitrosoguanidine and methylmethanesulfonic acid, ultraviolet irradiation, or treatment with a mutation agent. It can be obtained by the natural mutation obtained.
- microorganisms with high acetaldehyde-degrading ability from a large number of microbial species and recognized strong activity against many acetic acid bacteria, gluconic acid bacteria and Pseudomonas bacteria.
- these microorganisms exhibit the ability to produce acetaldehyde in the presence of ethanol, and that ingredients in foods such as beer and ingredients in saliva further increase such acetaldehyde producing activity.
- the microorganism used in the composition according to the present invention can be identified as follows, for example.
- a microorganism that specifically degrades acetaldehyde without producing acetaldehyde from ethanol is searched.
- a strain satisfying these requirements can be obtained by screening as follows as an example. 1) Applying fruit epidermis washing liquid, fermented food, human intestinal contents, etc. to an appropriate selective medium, 2) isolate the grown colonies, and 3) efficiently decompose acetaldehyde without producing acetaldehyde in the presence of ethanol. Select possible microbial strains.
- a microorganism capable of decomposing acetaldehyde without producing acetaldehyde from ethanol even in a liquor / saliva mixed system is obtained.
- the presence or absence of hydrogen peroxide during the acetaldehyde decomposition reaction is confirmed by a colorimetric method using peroxidase.
- aldehyde-degrading microorganisms such as nonenal and hexanal, which are odor-causing substances, can also be used.
- the microorganism can be identified according to the screening for microorganisms having the acetaldehyde-degrading activity.
- the medium used for the growth of the microorganism used in the present invention may contain, for example, a carbon source (for example, glucose etc.) and a nitrogen source that can be assimilated by the microorganism.
- a carbon source for example, glucose etc.
- a nitrogen source that can be assimilated by the microorganism.
- the nitrogen source include organic nitrogen sources (peptone, meat extract, yeast extract, corn, steeper, liquor, etc.), inorganic nitrogen sources (ammonium sulfate, ammonium chloride, etc.) and the like.
- the medium may contain salts composed of a cation such as sodium ion, potassium ion, calcium ion and magnesium ion and an anion such as sulfate ion, chlorine ion and phosphate ion.
- the culture medium can also contain trace elements such as vitamins and acids.
- concentration of the carbon source in the medium is, for example, about 0.1% to 10%.
- concentration of the nitrogen source in the medium varies depending on the type, but is, for example, about 0.01% to 5%.
- concentration of inorganic salts in the medium is, for example, about 0.001% to 1%.
- acetaldehyde is added to the medium for the purpose of removing contamination with microorganisms that cannot decompose acetaldehyde.
- concentration of acetaldehyde to be added can be appropriately determined, but is preferably 0.1%.
- the growth conditions of the microorganism used in the present invention are, for example, 1 to 3 days (preferably 2) at a temperature of 20 ° C. to 30 ° C. (preferably 25 to 28 ° C.) and a pH of 5.0 to 7.0 (preferably pH of 6.0 to 6.5). Day).
- the microbial cells may be used for the composition according to the present invention in a state suspended in the culture solution after culturing, or collected or concentrated from the culture solution by a usual method such as centrifugation. It may be used.
- the recovered or concentrated cells can be immobilized on a suitable carrier and used as immobilized cells. Or you may use the collect
- the composition according to the present invention can be made into a dosage form such as a powder, a tablet, or a capsule by mixing the bacterial cell powder and various excipients, for example.
- an extract such as a cell extract or an enzyme extract fraction or a disrupted cell body (for example, one containing a cell membrane) is prepared from a microorganism by a conventional method such as ultrasonic disruption or surfactant treatment, and the present invention. It can also be used for the composition which concerns on.
- extracts and disrupted bacterial cells may be used as they are, or may be immobilized on an appropriate carrier and used as an immobilized enzyme.
- the composition according to the present invention may be taken so as to take, for example, 5 to 80 mg, preferably 10 to 60 mg, of dry microbial cells of the contained microorganism. it can.
- the food or drink according to the present invention contains the composition according to the present invention.
- the microorganism according to the present invention by adding the microorganism according to the present invention to a food or drink, it can be a food or drink that reduces the carcinogenic risk of upper gastrointestinal tract cancer or a food and drink that prevents the ripening odor in the oral cavity.
- the food and drink according to the present invention are, for example, live cells, dead cells, immobilized cells, cell extracts, disrupted cells, acetaldehyde-degrading enzyme preparations purified from microorganisms, etc. Containing.
- acetaldehyde present or generated in the body can be removed by ingesting the composition according to the present invention.
- a sweetener, a sour agent, a fragrance, an antioxidant, and the like may be appropriately added to the food and drink according to the present invention.
- Specific product forms include lozenges, gums, candies, tablets, powders, drinks, yogurts and the like.
- the method of adding additives and the method of processing into foods or beverages can be selected and implemented as appropriate in consideration of the properties of the product.
- the enzyme solution (dialysis internal solution) 50 ⁇ L and beverages (green tea, oolong tea, beer, whiskey) 550 ⁇ L were mixed and incubated at 37 ° C. for 10 minutes.
- an aqueous solution (3 ppm AcH / 5% EtOH) containing acetaldehyde (3 ppm) and ethanol (5%) at concentrations originally contained in beer was also tested.
- the reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution.
- the acetaldehyde concentration in the reaction solution was measured by a headspace gas chromatograph method.
- Acetaldehyde concentration measurement by headspace gas chromatographic method SHIMADZU GC-2010 model using Perkin Elmer TurboMatrix 40 type headspace autosampler and equipped with INNOWAX 19091N-233 capillary column (length 30m, inner diameter 0.25mm, film 0.25 ⁇ m) This was done by using a gas chromatograph.
- acetaldehyde (AcH) measurement result is shown in FIG.
- FIG. 1 particularly significant acetaldehyde production was observed in beer.
- JCM7642 strain is known as a microorganism with acetaldehyde-degrading ability, but the result is that when this bacterium is taken after drinking beer, the concentration of acetaldehyde in the oral cavity is rather increased due to the strong ethanol oxidation activity that coexists in the microorganism cells. It indicates that there is a risk of doing.
- the cells of one platinum loop were suspended in 50 ⁇ L of physiological saline. This bacterial cell suspension (50 ⁇ L) was added to 50 mM phosphate buffer (550 ⁇ L) containing 5% ethanol (final concentration), and incubated at 37 ° C. for 10 minutes. The reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution. Acetaldehyde contained in the reaction solution was measured by the method described in Comparative Example 1.
- acetaldehyde (AcH) measurement results are shown in FIG. As shown in FIG. 2, all the microorganisms produced acetaldehyde of 2 ppm or more.
- Gluconobacter genus bacteria are generally known as microorganisms with acetaldehyde-degrading ability, but the result is that, when these microorganisms are taken after ingestion of alcoholic beverages, the strong ethanol oxidation activity that coexists in these microorganism cells causes the oral cavity This indicates that there is a risk that the concentration of acetaldehyde will rather increase.
- the cells of one platinum loop were suspended in 50 ⁇ L of physiological saline. This cell suspension (50 ⁇ L) was added to 50 mM phosphate buffer (550 ⁇ L) containing 5% ethanol (final concentration), and incubated at 37 ° C. for 10 minutes. The reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution. Acetaldehyde contained in the reaction solution was measured by the method described in Comparative Example 1.
- the cells of the microorganism produced 17.8 ppm of acetaldehyde.
- the results show that if you take this microorganism after ingesting an alcoholic beverage, there is a risk that the concentration of acetaldehyde in the oral cavity will rather increase due to the strong ethanol oxidation activity that coexists in these microbial cells. .
- Example 1 Reaction of microbial cells of Gluconobacter microorganisms and beer G. frateurii JCM20278, G. kondonii NBRC3266 and G. kanchanaburiensis NBRC103587 were cultured in the same manner as described in Comparative Example 1. . Next, these three microorganisms and the five Gluconobacter microorganisms described in Comparative Example 2 were suspended in 500 ⁇ L of MilliQ water so that the optical turbidity at 600 nm was about 0.9.
- Example 2 Removal of acetaldehyde in saliva using freeze-dried cells of Gluconobacter kondonii G. kondonii NBRC3266 was cultured by the method described in Example 1, and the resulting cells were subjected to freeze-drying. About 30 mg of freeze-dried cells were suspended in 500 ⁇ L of MilliQ water.
- 50 ⁇ L of the cell suspension was added to a beer / saliva mixed solution (volume ratio: 1: 1, 550 ⁇ L) containing acetaldehyde (initial concentrations of 2.5 ppm and 9.5 ppm) and incubated at 37 ° C. for 10 minutes.
- 50 ⁇ L of the same cell suspension was added to 550 ⁇ L of 0.1% ethanol and incubated at 37 ° C. for 10 minutes.
- the reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution.
- the acetaldehyde contained in the reaction solution was measured by the method described in Comparative Example 1.
- acetaldehyde (AcH) measurement results are shown in FIG. As shown in FIG. 3, acetaldehyde was not produced even in 0.1% ethanol or beer / saliva mixed system, and acetaldehyde in the beer / saliva mixed system was efficiently decomposed by freeze-dried cells.
- the reaction solution (0.6 mL each) obtained in the same manner as above was added to 0.05 mL of 20 mM 2,4-dichlorophenol and 20 mM 4-amino Antipyrine 0.05 mL, water 0.2 mL, and 10 mg / mL horseradish peroxidase 0.1 mL were added, and the mixture was incubated at room temperature for 20 minutes until the absorbance at 505 nm showed a constant value.
- the concentration of hydrogen peroxide contained in the reaction solution was estimated from the increase in absorbance at 505 nm. As a result, the hydrogen peroxide concentration in all the reaction solutions was below the detection limit of 0.5 ⁇ M (0.02 ppm) of this method.
- Example 3 Time course and specific activity of acetaldehyde removal G. kondonii NBRC3266 was cultured by the method described in Example 1, and the obtained cells were subjected to lyophilization. 10 mg of freeze-dried cells were suspended in 500 ⁇ L of MilliQ water.
- a 1 1 mixture of beer and 50 mM potassium phosphate buffer (pH 7.0) was prepared, and acetaldehyde was added to 7.8 ppm. 50 ⁇ L of cell suspensions having different cell concentrations were added to 550 ⁇ L of this mixed solution (5 in total) and incubated at 37 ° C. After reacting for 0, 1, 3, 5 and 10 minutes, the reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution. Acetaldehyde contained in the reaction solution was measured by the method described in Comparative Example 1.
- Example 4 Effect of the amount of cells on the removal of acetaldehyde G. kondonii NBRC3266 was cultured by the method described in Example 1, and the resulting cells were subjected to lyophilization. 10 mg of freeze-dried cells were suspended in 500 ⁇ L of MilliQ water. This was further diluted with MillQ water to prepare suspensions with different cell concentrations.
- a 25 mM potassium phosphate buffer (pH 7.0) containing 2.5% ethanol was prepared to dissolve acetaldehyde.
- 50 ⁇ L of the cell suspension adjusted to each cell concentration was added to 550 ⁇ L of this mixed solution, and reacted at 37 ° C. for 1 minute or 5 minutes.
- the initial acetaldehyde concentrations in the 1 minute and 5 minute reactions were 10.1 ppm and 11.1 ppm, respectively.
- the reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution.
- Acetaldehyde contained in the reaction solution was measured by the method described in Comparative Example 1.
- acetaldehyde (AcH) measurement results are shown in FIG. In FIG. 4, open circles indicate a reaction for 1 minute, and black circles indicate a reaction for 5 minutes.
- acetaldehyde can be removed by using 0.5 mg of cells in a 5-minute reaction and 1.0 mg of cells in a 1-minute reaction. From this example, the acetaldehyde removal rate of the cells was estimated to be about 3.9 ⁇ g ⁇ min ⁇ 1 ⁇ (mg dry cells) ⁇ 1 .
- Example 5 Removal of intermittently added acetaldehyde G. kondonii NBRC3266 was cultured by the method described in Example 1, and the obtained cells were subjected to lyophilization. 30 mg of freeze-dried cells were suspended in 500 ⁇ L of MillQ water.
- a 25 mM potassium phosphate buffer solution (pH 7.0) containing 1.5% ethanol was prepared, and acetaldehyde was dissolved to about 10 ppm and kept at 37 ° C.
- a cell suspension (200 ⁇ L) was added to this reaction system (1.80 mL), and when incubated at 37 ° C. for 3 minutes, 200 ⁇ L was sampled from the reaction system, mixed with 20 ⁇ L of 6M perchloric acid, The reaction was stopped.
- the acetaldehyde contained in each stopped reaction solution was measured by the method described in Comparative Example 1.
- a control experiment was carried out using the same procedure except that the addition of aqueous acetaldehyde (200 ⁇ L) after sampling of the reaction solution was replaced with the addition of water (200 ⁇ L).
- Example 6 Effect of Ethanol Concentration on Acetaldehyde Removal G. kondonii NBRC3266 was cultured by the method described in Example 1, and the resulting cells were subjected to lyophilization. 10 mg of freeze-dried cells were suspended in 500 ⁇ L of MilliQ water.
- a 25 mM potassium phosphate buffer (pH 7.0) containing ethanol at each concentration was prepared, and 6 to 8 ppm (final concentration) acetaldehyde was dissolved.
- a cell suspension (50 ⁇ L) was added to 550 ⁇ L of this mixed solution, and reacted at 37 ° C. for 5 minutes. The reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution. Acetaldehyde contained in the reaction solution was measured by the method described in Comparative Example 1.
- the bacterial cells efficiently decomposed almost all acetaldehyde in the reaction system containing ethanol up to a concentration of 10% (acetaldehyde residual amount: 0.07 ppm to 0.19 ppm).
- the reaction system containing ethanol at a concentration higher than 20% has reduced AcH decomposition ability, and the residual acetaldehyde in the system containing 20%, 35% and 50% ethanol is 3.4 ppm, 5.5 ppm and 5.2 ppm.
- Example 7 Effect of pH on removal of acetaldehyde G. kondonii NBRC3266 was cultured by the method described in Example 1, and the resulting cells were subjected to lyophilization. 10 mg of freeze-dried cells were suspended in 500 ⁇ L of MilliQ water.
- Buffers with various pH containing 2.5% ethanol were prepared, and 10.5 ⁇ 0.5 ppm (final concentration) acetaldehyde was dissolved.
- a cell suspension (50 ⁇ L) was added to 550 ⁇ L of this mixed solution, and reacted at 37 ° C. for 5 minutes. The reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution. Acetaldehyde contained in the reaction solution was measured by the method described in Comparative Example 1.
- FIG. 6 shows the residual concentration of acetaldehyde when acetaldehyde having an initial concentration of 10.5 ⁇ 0.5 ppm is decomposed by Gluconobacter cells.
- the microbial cells showed the ability to decompose acetaldehyde over a wide range of pH 2-10. Sufficient degradation activity was observed especially in the acidic region, suggesting the possibility that the microbial cells show excellent AcH resolution in the stomach and intestines.
- the amount of bacterial cells necessary for removing acetaldehyde in the oral cavity after drinking is estimated as follows.
- the rate of acetaldehyde degradation per mg of freeze-dried cells of G. kondonii NBRC3266 is 4-5 ⁇ g ⁇ min ⁇ 1 ⁇ (mg dry cells) ⁇ 1 .
- the concentration of acetaldehyde in saliva after drinking is often reported to be in the range of 2-4 ppm (Yokoyama A, Tsutsumi E, Imazeki H, Suwa Y, Nakamura C, Mizukami T, Yokoyama T. Alcohol Clin. Exp. Res. 32, 1607-1614 (2008)).
- a standard saliva amount (stimulated saliva amount) secreted after chewing gum for 10 minutes has been shown to be 11 ml (Miwa et al., Journal of Japanese Society of Oral Examinations 1, 40-43 (2009)) This shall be regarded as the amount of saliva present in the oral cavity. If 10 ppm acetaldehyde (110 ⁇ g) is present in 11 ml of saliva in the oral cavity after drinking, acetaldehyde can be eliminated within 1 minute if 30 mg of lyophilized cells are present in the oral cavity. It is expected to be.
- Example 8 Removal of odorous substances G. kondonii NBRC3266 was cultured by the method described in Example 1, and the obtained cells were subjected to lyophilization. 10 mg of freeze-dried cells were suspended in 500 ⁇ L of MilliQ water.
- the cell suspension (50 ⁇ L) was added to 550 ⁇ L of 25 mM potassium phosphate buffer (pH 7.0) containing 0.5 ppm nonenal or 0.5 ppm hexanal, and reacted at 37 ° C. for 5 minutes.
- the reaction was stopped by adding 60 ⁇ L of 6M perchloric acid to the reaction solution.
- Nonenal or hexanal contained in the reaction solution was measured by the same method as described in Comparative Example 1.
- the bacterial cells efficiently decomposed these aldehydes, and nonenal or hexanal could not be detected in the reaction solution after the reaction was stopped.
- Example 9 Effect of ingestion of Gluconobacter condonii cells on acetaldehyde removal in the oral cavity after drinking Alcohol (51 years old, weight 70 kg; ALDH2 * 1 / * 1) has breakfast (bread, vegetables, milk) I took it for 11 hours at 6am. Before drinking, chewing gum for saliva collection was sampled and 2 ml of saliva was sampled.
- Whiskey (Yamazaki 10 years, alcohol content 40 degrees) was diluted with mineral water to prepare water split (alcohol content 13 degrees; acetaldehyde concentration 18.2 ppm). By taking 90 ml of this water split over 8 minutes and repeating this 4 times, a total of 360 ml of water split was taken over 32 minutes (total alcohol consumption: 47 kg; 0.67 g / kg body weight). During this time, I did not take a side meal.
- the gluconobacter condonii cells (freeze-dried product 30 mg) were immediately taken after the first saliva collection and uniformly dispersed in the oral cavity.
- chewing gum for collecting saliva was chewed 5 minutes after taking, and 2 ml of saliva was collected together with the cells.
- chewing gum for saliva collection was chewed and 2 ml of saliva was collected.
- saliva was collected 15 minutes after the completion of drinking.
- the specific activity of the Gluconobacter chondoni cells taken was 4 U / mg.
- FIG. 7 The transition of oral acetaldehyde concentration after drinking is shown in FIG. In FIG. 7, the horizontal axis indicates the time (min) after drinking. Immediately after drinking, a pulsed acetaldehyde concentration increase of about 2.5 ppm was observed. When Gluconobacter condonii cells were not taken (“control” graph in FIG. 7), the acetaldehyde in the oral cavity remained at a concentration of 1.8 ppm over about 1 hour, 1.1 ppm after 90 minutes, 0.9 ppm after 120 minutes, After 180 minutes, it became 0.6 ppm.
- the acetaldehyde in the oral cavity immediately decreased to 0.5 ppm immediately after taking, and increased again after spitting out or swallowing the cells. After 30 minutes of completion, it became 1.4 ppm, after 1 hour, 1.1 ppm, and after 90 minutes, 0.8 ppm.
- the concentration of oral acetaldehyde was not significantly different from 120 minutes after drinking and 180 minutes after drinking without Gluconobacter condonii cells, but up to 90 minutes after drinking depending on the consumption of Gluconobacter condonii cells Oral acetaldehyde concentration was significantly lower than non-dose.
- the formulation of the gluconobacter condonii cells in the oral cavity can be maintained more effectively at a low level if the device formulation is used to establish gluconobacter chondoni cells in the oral cavity for a longer period of time.
- Example 10 Effect of adding Gluconobacter condonii cells to foods G.
- kondonii NBRC3266 was cultured by the method described in Example 1, and the resulting cells were subjected to lyophilization.
- generation by the combination of the obtained microbial cell and the foodstuff shown in following Table 1 was investigated.
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Abstract
Description
グルコン酸培地(2%グルコン酸ナトリウム、0.3%グルコース、0.3%酵母エキス、0.2%ペプトン)50mLを用いて、Gluconobacter oxydans JCM7642株を30℃で3日間前培養した。次いで、前培養後の培養液10mLを同組成の培地1Lに加え、30℃で3日間培養した。培養後、菌体は遠心分離(3,000×g)により回収した。
5種のGluconobacter属微生物(グルコノバクター・アサイ(G. asaii)JCM21279、グルコノバクター・セリナス(G.cerinus)JCM20277、グルコノバクター・エスピー(Gluconobacter sp.)JCM20280、グルコノバクター・タイランディカス(G. thailandicus)JCM12310及びG.oxydans JCM7642)を、アセトバクター寒天培地(10%グルコース、1%酵母エキス、3%炭酸カルシウム、1.5%寒天)上で30℃で3日間培養した。
アセトアルデヒド分解微生物として知られるPseudomonas sp. AL-5株を培地(1.5%寒天)上で30℃で3日間培養した。
グルコノバクター・フラツリイ(G. frateurii)JCM20278、G. kondonii NBRC3266及びG. kanchanaburiensis NBRC103587を比較例1で述べた方法と同様にして培養した。次いで、これら3種の微生物並びに比較例2で述べた5種のGluconobacter属微生物を、それぞれ600nmにおける光学濁度が約0.9となるように500μLのMilliQ水に懸濁した。
G. kondonii NBRC3266を実施例1で述べた方法により培養し、得られた菌体を凍結乾燥に供した。凍結乾燥菌体約30mgをMilliQ水500μLに懸濁した。
G. kondonii NBRC3266を実施例1で述べた方法により培養し、得られた菌体を凍結乾燥に供した。凍結乾燥菌体10mgをMilliQ水500μLに懸濁した。
G. kondonii NBRC3266を実施例1で述べた方法により培養し、得られた菌体を凍結乾燥に供した。凍結乾燥菌体10mgをMilliQ水500μLに懸濁した。これをさらにMillQ水で希釈することにより異なる菌体濃度の懸濁液を調製した。
G. kondonii NBRC3266を実施例1で述べた方法により培養し、得られた菌体を凍結乾燥に供した。凍結乾燥菌体30mgをMillQ水500μLに懸濁した。
G. kondonii NBRC3266を実施例1で述べた方法により培養し、得られた菌体を凍結乾燥に供した。凍結乾燥菌体10mgをMilliQ水500μLに懸濁した。
G. kondonii NBRC3266を実施例1で述べた方法により培養し、得られた菌体を凍結乾燥に供した。凍結乾燥菌体10mgをMilliQ水500μLに懸濁した。
G. kondonii NBRC3266を実施例1で述べた方法により培養し、得られた菌体を凍結乾燥に供した。凍結乾燥菌体10mgをMilliQ水500μLに懸濁した。
被験者(51歳、体重70 kg;ALDH2*1/*1)は、朝食(食パン、野菜、牛乳)を朝6時にとり、11時間絶食した。飲酒前に唾液採取用のチューインガムを噛み、唾液2 mlをサンプリングした。
G. kondonii NBRC3266を実施例1で述べた方法により培養し、得られた菌体を凍結乾燥に供した。得られた菌体と下記の表1に示す食品との組み合わせによるアセトアルデヒド(AcH)生成の可能性を調べた。
Claims (12)
- アルデヒド分解活性を有するグルコノバクター属に属する微生物又はその抽出物若しくは菌体破砕物を含有するアルデヒド除去組成物であって、前記グルコノバクター属に属する微生物はアルデヒド生成活性よりもアルデヒド分解活性が高いことを特徴とする、前記アルデヒド除去組成物。
- 前記グルコノバクター属に属する微生物がアルデヒド濃度と比較してアルコール濃度が高い条件下でアルデヒドを分解できることを特徴とする、請求項1記載のアルデヒド除去組成物。
- 前記グルコノバクター属に属する微生物がグルコノバクター・コンドニイ(Gluconobacter kondonii)又はグルコノバクター・カンチャナブリエンシス(Gluconobacter kanchanaburiensis)に属する微生物である、請求項1記載のアルデヒド除去組成物。
- 前記グルコノバクター・コンドニイ又はグルコノバクター・カンチャナブリエンシスに属する微生物がNBRC3266又はNBRC103587で特定される微生物である、請求項3記載のアルデヒド除去組成物。
- 前記アルデヒドがアセトアルデヒドである、請求項1記載のアルデヒド除去組成物。
- 前記アルコールがエタノールである、請求項2記載のアルデヒド除去組成物。
- 上部消化管癌罹患リスク低減組成物である、請求項1記載のアルデヒド除去組成物。
- 上部消化管癌が口腔癌、咽頭癌及び食道癌から成る群より選択される、請求項7記載のアルデヒド除去組成物。
- 臭い除去組成物である、請求項1記載のアルデヒド除去組成物。
- 臭いが熟柿臭である、請求項9記載のアルデヒド除去組成物。
- 飲酒後の口腔内アセトアルデヒド除去に使用するための、請求項1記載のアルデヒド除去組成物。
- 請求項1~11のいずれか1項記載のアルデヒド除去組成物を含有する飲食品。
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US13/698,331 US20130058887A1 (en) | 2010-05-18 | 2011-05-13 | Aldehyde-removing composition |
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Cited By (6)
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CN102526166A (zh) * | 2012-01-21 | 2012-07-04 | 赵庭义 | 一种治疗小儿口疮的外用药物及其制备方法 |
JP2015199701A (ja) * | 2014-03-31 | 2015-11-12 | キユーピー株式会社 | 酢酸菌由来アルデヒド脱水素酵素含有物を配合した化粧料及び加齢臭抑制剤 |
JP2015205862A (ja) * | 2014-04-09 | 2015-11-19 | キユーピー株式会社 | 化粧料及び加齢臭抑制剤 |
JP2016160203A (ja) * | 2015-02-27 | 2016-09-05 | キユーピー株式会社 | 化粧料及び加齢臭抑制剤 |
JP2017075130A (ja) * | 2015-10-16 | 2017-04-20 | サッポロホールディングス株式会社 | アセトアルデヒド除去剤 |
JP2019083771A (ja) * | 2017-11-08 | 2019-06-06 | アサヒビール株式会社 | 微生物培養用培地及び酢酸菌の検出方法 |
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Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
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CN102526166A (zh) * | 2012-01-21 | 2012-07-04 | 赵庭义 | 一种治疗小儿口疮的外用药物及其制备方法 |
JP2015199701A (ja) * | 2014-03-31 | 2015-11-12 | キユーピー株式会社 | 酢酸菌由来アルデヒド脱水素酵素含有物を配合した化粧料及び加齢臭抑制剤 |
JP2015205862A (ja) * | 2014-04-09 | 2015-11-19 | キユーピー株式会社 | 化粧料及び加齢臭抑制剤 |
JP2016160203A (ja) * | 2015-02-27 | 2016-09-05 | キユーピー株式会社 | 化粧料及び加齢臭抑制剤 |
JP2017075130A (ja) * | 2015-10-16 | 2017-04-20 | サッポロホールディングス株式会社 | アセトアルデヒド除去剤 |
JP2019083771A (ja) * | 2017-11-08 | 2019-06-06 | アサヒビール株式会社 | 微生物培養用培地及び酢酸菌の検出方法 |
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JPWO2011145518A1 (ja) | 2013-07-22 |
JP5637209B2 (ja) | 2014-12-10 |
US20130058887A1 (en) | 2013-03-07 |
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