JP2003033195A - Method for producing tetrahydrocurcumin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing tetrahydrocurcumins and a method for producing a composition containing tetrahydrocurcumins using microbial cells, cultures or processed products thereof. SOLUTION: In the presence of an O / W emulsion, microbial cells, a culture solution or a processed product thereof having the activity of converting curcumins into tetrahydrocurcumins are converted into curcumins or compositions containing curcumins. A method for producing a tetrahydrocurcumin or a composition containing the tetrahydrocurcumin, the method comprising causing the compound to act.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing a tetrahydrocurcumin or a composition containing a tetrahydrocurcumin.

[0002]

BACKGROUND OF THE INVENTION Curcumins are well known as yellow pigments contained in tropical plants belonging to the genus Turmeric of the ginger family. Turmeric rhizome powder (turmeric) and its purified product, curcumin, are used as flavoring agents and coloring agents in foods. Curcumins are known to have medicinal effects such as an antioxidant effect, an anti-inflammatory effect, a cholesterol-reducing effect, and a carcinogenesis suppressing effect, and are used as safe medicinal components derived from foods.

Tetrahydrocurcumin (THU1) is
It is known to have stronger antioxidant activity than curcumin [JP-A-2-49747 and JP-A-2].
-51595, Bioscience, Biotechnology and Biochemistry (Biosci. Biotec
h. Biochem.), 59 , 1609 (1995)]. Further, since tetrahydrocurcumin is colorless and odorless, it is a material that overcomes the drawbacks of curcumin, especially when coloring is a problem.

As a method for producing tetrahydrocurcumin, a method for producing curcumin by hydrogenation using a metal catalyst is known. However, the use of the synthetic tetrahydrocurcumin produced by this method for food and drink has a problem in food hygiene. Suitable methods for producing edible tetrahydrocurcumins include:
A method for producing tetrahydrocurcumins by mixing curcumin with microbial cells and the like and keeping the mixture warm (Japanese Patent Application Laid-Open No. H11-115242)
No. 235192), but further improvement in yield is required.

The solubility of curcumin, which is a typical curcumin, in water is as low as 0.1 μg / ml or less at room temperature. When conducting a biochemical reaction using microbial cells in an aqueous system,
Curcumins, which are poorly soluble in water, are used by first dissolving them in an organic solvent that dissolves curcumins such as ethanol.However, if curcumin precipitates in the reaction solution or if the amount of organic solvent used is increased, microbial cells become inactive. However, it is difficult to react efficiently. In addition, some organic solvents exhibit toxicity, and use of the reaction product in the food and drink field may be limited.

Curcumin, which is sparingly soluble in water, can be well dispersed in an aqueous system without using an organic solvent, and the equilibrium between the reaction of forming the target product and the decomposition reaction of the target product can be inclined toward the former, and the recovery of the target product is easy. It is desired that such a technique be used. The emulsification technology is a method often used in food and drink and pharmaceutical fields because it disperses a poorly water-soluble compound in water.
For use in structural modification reactions, the combination of enzyme and product shows that alkaline phosphatase and p-nitropenylphosphat
e) [Indian Journal of Biochemistry & Biophysics, 3
2 , 261 (1995)], lipase and 16-hydroxyhexadecanoic acid [Biochemica e
Biophysica Acta, 1257 , 239 (1995)], bilirubin oxidase and bilirubin [European Journal of Bioche
mistry, 183 , 347 (1989)], and phospholipase A2 and phosphatidylcholine [Colloids and Surfa
ces, 128 , 17 (1997)], but no report on the structural conversion reaction of curcumins using microbial cells.

[0007]

DISCLOSURE OF THE INVENTION An object of the present invention is to provide a method for producing tetrahydrocurcumins using a microbial cell, a culture, or a treated product thereof, and a method for producing a composition containing tetrahydrocurcumins. Especially.

[0008]

The present invention includes the following (1).
About (9). (1) Formula (I) in the presence of an O / W emulsion

[0009]

[Chemical 3]

[In the formula (I), R ¹ , R ² , R ³ and R ⁴
Are the same or different and represent hydrogen, hydroxy or lower alkoxy], and a curcumin or a curcumin derivative (hereinafter simply referred to as curcumin) is represented by the formula (II)

[0011]

[Chemical 4]

[In the formula (II), R ¹ , R ² , R ³ and R
⁴ are the same or different and each represent hydrogen, hydroxy or lower alkoxy]], and a microbial cell, a culture solution or a microbial cell having an activity of converting to tetrahydrocurcumin or a tetrahydrocurcumin derivative (hereinafter simply referred to as tetrahydrocurcumins) A method for producing a tetrahydrocurcumin or a composition containing a tetrahydrocurcumin, which comprises causing the treated product of 1. to act on a curcumin or a composition containing a curcumin.

(2) The production method according to (1), wherein the curcumin or the composition containing the curcumin is contained in an O / W emulsion. (3) The production method according to (1) or (2), wherein the O / W type emulsion is obtained by mixing an oil, an emulsifier or a surfactant and an aqueous medium. (4) The average particle size of the O / W emulsion is 10 n
The method according to any one of (1) to (3), which is m to 10 μm.

(5) The method according to any one of (1) to (4), wherein the curcumin-containing composition is a composition obtained by treating a plant belonging to the genus Turmeric. (6) The microorganism is a microorganism belonging to the genera Devalyomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, Lactobacillus, Staphylococcus, or Pediococcus (1) to (5). The method described in any one of.

(7) A curcumin, a microbial cell, a culture solution or a treated product of the microorganism having an activity of converting curcumin to tetrahydrocurcumin in the presence of oils and an emulsifier or a surfactant in an aqueous medium. Alternatively, the method for producing a tetrahydrocurcumin or a composition containing a tetrahydrocurcumin is characterized by acting on a composition containing a curcumin.

(8) The method according to (7), wherein the curcumin-containing composition is a composition obtained by treating a plant belonging to the genus Turmeric. (9) The microorganism is a microorganism belonging to the genera Devalyomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, Lactobacillus, Staphylococcus or Pediococcus (7) or (8). The method described in.

[0017]

BEST MODE FOR CARRYING OUT THE INVENTION The alkyl moiety of the lower alkoxy of R ¹ , R ² , R ³ and R ⁴ in formula (I) and formula (II) is a straight or branched alkyl group having 1 to 6 carbon atoms. , For example, methyl, ethyl, propyl, isopropyl,
Butyl, sec-butyl, tert-butyl, pentyl, hexyl and the like can be mentioned. As the alkoxy, methoxy and ethoxy having 1 to 2 carbon atoms are preferable. Formula (I)
In the above, as R ² and R ⁴ , a compound which is hydroxy is preferable.

Specific curcumins include, for example, U
1 [diferuloylmethane,
(E, E) -1,7-bis (4-hydroxy-3-methoxyphenyl) -1,6-heptadiene-3,5-dione, sometimes hereinafter simply referred to as "curcumin"], U2
[Demethoxycurcumin, 4-
Hydroxycinnamoyl (feruloyl) methane], U
3 [bisdimethoxycurcumin
in), bis (4-hydroxycinnamoyl) methane], DMU1
[(E, E) -1,7-bis (3,4-dimethoxyphenyl) -1,6-heptadien
e-3,5-dione], DHU1 [(E, E) -1,7-Bis (3,4-dihydro
xyphenyl) -1,6-heptadiene-3,5-dione] and the like,
U1, U2, U3 and DHU1 are preferably used, U
1 and DHU1 are particularly preferably used.

The method for producing the O / W emulsion used in the present invention is not particularly limited, but it can be produced, for example, by mixing an oil, an emulsifier or a surfactant and an aqueous medium. The mixing ratio is not particularly limited, but, for example, to 1 part by weight of oils, 0.01 emulsifier or surfactant
10 to 10 parts by weight, preferably 1 to 100 parts by weight of the aqueous medium,
More preferably, the emulsifier or surfactant is 0.1 to 1 part by weight, and the aqueous medium is 2 to 50 parts by weight.

The method for preparing the emulsion is not particularly limited, and a stirrer such as a propeller mixer or a turbine mixer,
High-speed rotating rotor disk type emulsifier such as colloid mill, high-speed rotating dispersion emulsifier such as homomixer, high-pressure high-speed injection type emulsifier such as jet homogenizer, ultrasonic wave, high-pressure homogenizer, vortex mixer, magnetic stirrer, preferably Can be prepared by using a high pressure homogenizer.

As a method for producing an O / W emulsion, for example, 0.1 to 1 part by weight of fats and oils, 0.1 to 1 part by weight of an emulsifier and 1 to 10 parts by weight of an aqueous medium are mixed to prepare about 40 to 6 parts.
1,000 to 20,000 with a mixer while keeping the temperature at 0 ℃
A method of stirring at 0 rpm for 1 to 10 minutes can be mentioned. O
The average particle size of the / W emulsion is not particularly limited,
10 nm to 10 μm is preferable, 50 nm to 5 μm is more preferable, and 100 nm to 2 μm is particularly preferable.

The O / W type emulsion used in the present invention may include curcumins or compositions containing curcumins. As a specific method for preparing an O / W emulsion containing curcumin or a composition containing curcumin, for example, 1 part by weight of curcumin and 2 to 1,000 parts by weight of oil, preferably 5 to 1 are used.
It is added to 100 parts by weight, and is 0 to 300 ° C, preferably 50.
Heat and melt at ˜200 ° C. to prepare curcumin-containing oils. The curcumin-containing oils can be used in the same manner as in the emulsion preparation method described above to produce an O / W emulsion containing curcumin or a composition containing curcumin.

The oils are not particularly limited as long as they can dissolve curcumins, and examples thereof include soybean oil, sesame oil,
Vegetable oil such as safflower, cottonseed oil, coconut oil, coconut oil, diacylglycerol-containing vegetable oil, long chain fatty acid triglyceride or medium chain fatty acid triglyceride (MC
T), α-tocopherol, saturated fatty acids, unsaturated fatty acids, cholesterol and the like.

Examples of emulsifiers or surfactants include lecithin emulsifiers, saponin emulsifiers, polysaccharide emulsifiers, protein emulsifiers, other emulsifiers, and synthetic surfactants. Examples of the lecithin-based emulsifier include egg yolk lecithin, plant lecithin such as soybean-derived lecithin, enzyme-treated lecithin such as phosphatidylglycerol, and enzymatically-decomposed lecithin such as lysolecithin and phosphatidic acid.

Examples of saponin-based emulsifiers include quillaja saponin, enju saponin, soybean saponin, yucca.
Home etc. Examples of the polysaccharide emulsifier include gum arabic, carrageenan, xanthan gum, modified starch such as octenyl succinic starch, and propylene glycerol alginate.

Examples of protein-based emulsifiers include gluten partially decomposed products, vegetable protein partially decomposed products, gliadin, casein, sodium caseinate, whey protein, casein, egg white powder, soybean protein and the like. Examples of other emulsifiers include sphingolipid, plant sterol, animal sterol, bile powder, tomato glycolipid and the like.

Examples of the synthetic surfactant include polyether type polymer surfactant, glycerin fatty acid ester (glycerin fatty acid ester, glycerin acetic acid ester, glycerin acetic acid fatty acid ester, glycerin lactic acid fatty acid ester, glycerin cenoic acid fatty acid ester, glycerin succinic acid). Fatty acid ester, glycerin diacetyl acid tartaric acid fatty acid ester, polyglycerin fatty acid ester, etc.), sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, calcium stearoyl lactylate, oxyethylene higher fatty acid alcohol, sodium oleate, morpholine fatty acid salt, poly Examples thereof include oxyethylene higher aliphatic alcohol.

As the aqueous medium, for example, glycerol,
Pure water, a buffer solution or the like is used. As the buffer solution, Cla
rk-Lubs buffer, Sorensen buffer,
Kolthoff buffer, Michaelis buffer, Atkins-Patin buffer, Palit
zsch buffer, Mcllvaine buffer, Me
nzel buffer, Walpole buffer, Hast
Examples include ing-Sendroy buffer and Gomori buffer.

As the Clark-Lubs buffer,
Examples thereof include potassium chloride-hydrochloric acid, potassium hydrogen phthalate-hydrochloric acid, potassium hydrogen phthalate-sodium hydroxide, potassium dihydrogen phosphate-sodium hydroxide, potassium borate-sodium hydroxide and the like. Sorense
Examples of the n buffer include glycine sodium chloride-
Examples include hydrochloric acid, sodium glycine chloride-sodium hydroxide, sodium citrate-hydrochloric acid, sodium citrate-sodium hydroxide, borax-hydrochloric acid, borax-sodium hydroxide, potassium dihydrogen phosphate-dipotassium hydrogen phosphate. .

Examples of Kolthoff's buffer include potassium hydrogen citrate-citric acid, potassium hydrogen citrate-hydrochloric acid, potassium hydrogen citrate-sodium hydroxide, succinic acid-borax, potassium hydrogen citrate-borax, and phosphorus. Examples thereof include potassium dihydrogen acid-borax, borax-sodium carbonate, hydrochloric acid-sodium carbonate, disodium hydrogen phosphate-sodium hydroxide and the like.

Examples of the Michaelis buffer include tartaric acid-sodium tartrate, lactic acid-sodium lactate, acetic acid-sodium acetate, potassium dihydrogen phosphate-disodium hydrogen phosphate, ammonium chloride-ammonia water, and sodium diethylbarbiturate. Sodium-
Examples thereof include hydrochloric acid, sodium diethylbarbiturate-hydrochloric acid, dimethylglycine sodium-hydrochloric acid and the like.

Examples of the Atkins-Pantin buffer solution include potassium borate-sodium carbonate. Examples of the Palitsch buffer include sodium borate-borax and the like. Mc
Examples of the llvaine buffer include disodium hydrogen phosphate-citric acid and the like.

Examples of Menzel buffers include sodium carbonate-sodium hydrogen carbonate. W
Examples of the buffer for alpole include sodium acetate-hydrochloric acid and acetic acid-sodium acetate. Has
Examples of the buffer solution for towing-Sendroy include disodium hydrogen phosphate-potassium dihydrogen phosphate.

As the buffer solution of Gomori, for example, tris buffer solution such as 2,4,6-trimethylpyridine-hydrochloric acid or trisaminomethane-hydrochloric acid, 2-aminomethyl-1,
Examples include 3-propanediol-hydrochloric acid. In the present invention, the oils, emulsifiers or surfactants and aqueous media used for preparing the O / W type emulsion are preferably of natural origin, and are preferably edible ingredients.

In particular, medium-chain fatty acid triglyceride (MCT) is preferable as an oil used for preparing an O / W type emulsion, and a mixture of polyglycerin fatty acid ester and enzymatically decomposed lecithin is used as an emulsifier or a surfactant. It is preferable to use pure water as the aqueous medium. The method for producing a tetrahydrocurcumin or a composition containing a tetrahydrocurcumin of the present invention is a microbial cell, a culture solution or a microbial cell having an activity of converting a curcumin to a tetrahydrocurcumin in the presence of an O / W type emulsion. And the composition containing curcumin or a curcumin.

The method for producing a tetrahydrocurcumin or a composition containing a tetrahydrocurcumin of the present invention is a method for converting a curcumin into a tetrahydrocurcumin in an aqueous medium in the presence of an oil and an emulsifier or a surfactant. The method includes the step of allowing a bacterial cell, a culture solution or a processed product thereof having an active microorganism to act on curcumin or a composition containing curcumin.

The reaction solution is a culture solution of the microorganism, cells or treated products thereof, curcumin or a composition containing curcumin, and an O / W emulsion prepared by the above-mentioned method, if necessary, with water. Is prepared in addition to an aqueous medium such as. In addition, the reaction solution is prepared by adding cells of the microorganism, a culture solution or a treated product thereof, curcumin or a composition containing curcumin, an oil and an emulsifier or a surfactant to an aqueous medium.

For the production of tetrahydrocurcumin or a composition containing tetrahydrocurcumin, the reaction solution is, for example, 0 to 100 ° C., preferably 10 to 60 ° C., particularly preferably 22 to 45 ° C., and the pH is 2 to 11 , Preferably 3-9, particularly preferably 4-8, under the conditions of 0.01-
The temperature can be kept for 168 hours, preferably 0.5 to 72 hours.

The amount of the microbial cells, the culture solution or the processed product thereof in the reaction solution is not particularly limited as long as it has an activity of converting curcumins to tetrahydrocurcumins, but, for example, in terms of wet cell weight. 1 μg to 800 mg /
ml, preferably 10 to 500 mg / ml, is added to the reaction solution. The concentration of the curcumin or the composition containing the curcumin in the reaction solution is not particularly limited, but, for example, as the curcumin in the reaction solution is 0.001 to 500 m
g / ml, preferably 0.01 to 100 mg / ml.

The concentration of the O / W type emulsion in the reaction solution is not particularly limited, but is, for example, 0.01 to 990 mg / ml,
It is preferably added so as to be 1 to 100 mg / ml. The concentration of oils in the reaction solution is not particularly limited, but is, for example, 0.01 to 990 mg / ml, preferably 1 to 100 m.
Add to g / ml. The concentration of the emulsifier or surfactant in the reaction solution is not particularly limited, but may be, for example, 0.
01-990 mg / ml, preferably 1-100 mg /
Add to make up to ml.

The reaction liquid is, for example, 0.5 to 1000 parts by weight of microbial cells, culture liquid or treated product thereof,
Preferably 1 to 10 parts by weight, 0.5 to 1000 parts by weight, preferably 1 to 10 parts by weight of curcumin or a composition containing curcumin, and 0.
0001 to 1000 parts by weight, preferably 0.05 to 10
Examples include parts by weight.

When the curcumin or the composition containing the curcumin is contained in the O / W type emulsion, the tetrahydrocurcumin or the composition containing the tetrahydrocurcumin is produced by converting the curcumin into the tetrahydrocurcumin. The method comprises the step of acting a microbial cell, a culture solution, or a treated product thereof having a microorganism-converting activity on an O / W emulsion containing curcumin or a composition containing curcumin.

If necessary, an O / W type emulsion containing the bacterial cells of the microorganism, a culture solution or a treated product thereof and curcumin or a composition containing curcumin is added to an aqueous medium such as water to obtain a reaction solution. And the reaction solution is 0 to 10
0 ° C, preferably 10 to 60 ° C, particularly preferably 20 to
The reaction is carried out under conditions of 50 ° C. and pH of 2 to 11, preferably 3 to 9, and particularly preferably 4 to 0.01 for 0.01 to 168 hours, preferably 0.5 to 72 hours.

The amount of the microbial cells, the culture solution or the processed product thereof in the reaction solution is not particularly limited as long as it has an activity of converting curcumins to tetrahydrocurcumins, but, for example, in terms of wet cell weight. 1 μg to 800 mg /
ml, preferably 10 to 500 mg / ml, is added to the reaction solution. The concentration of the O / W type emulsion containing the curcumin or the composition containing the curcumin in the reaction solution is not particularly limited, but is, for example, 0.01 to 990 m.
g / ml is preferably added so as to be 1 to 100 mg / ml.

The concentration of curcumin or the composition containing curcumin in the O / W emulsion is not particularly limited, but, for example, 0.01 to 999 mg / ml, preferably 1 to 200 m as curcumin in the emulsion.
Preference is given to using emulsions which are g / ml.
The reaction liquid is, for example, 0.5 to 1,000 parts by weight, preferably 1 to 1000 parts by weight of microorganism cells, culture liquid or a treated product thereof.
The composition O / W type emulsion containing 10 parts by weight and curcumin or curcumin is 0.0001-100.
0 parts by weight, preferably 0.05 to 10 parts by weight, and the aqueous medium comprises 0 to 1000 parts by weight, preferably 10 to 100 parts by weight.

If necessary, a buffer solution, an emulsifier or a surfactant, an organic solvent, a coenzyme and the like can be present in the reaction solution. Examples of the buffer include sodium hydrogenphosphate, glycine, N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid, ammonium acetate, tris (hydroxymethyl) methylglycine, and the like in addition to the above-mentioned buffers. .

Examples of the emulsifier or surfactant include Triton X100 and the like in addition to the above-mentioned emulsifier or surfactant. Examples of the organic solvent include ethanol, methanol, glycerin and the like. Examples of coenzymes include beta nicotinamide adenine dinucleotide phosphate, beta nicotinamide adenine dinucleotide phosphate, reduced beta nicotinamide adenine dinucleotide, reduced beta nicotinamide adenine dinucleotide phosphate, and the like.

The tetrahydrocurcumins formed in the reaction solution or the composition containing the tetrahydrocurcumins can be used as they are as food and drink or feed.
You may refine and use it. The purification method is not particularly limited, and examples thereof include organic solvent extraction, centrifugation, column chromatography, freeze drying, recrystallization, hot air drying, and the like.

The products of curcumins and tetrahydrocurcumins can be detected by analysis by HPLC or the like.
For the reaction of curcumin or a composition containing curcumin with a microbial cell, a culture solution or a treated product thereof, for example, a container such as a wooden barrel is used without controlling temperature, aeration amount and hydrogen ion concentration. Although it can be carried out by leaving it for a certain period of time, it is preferably carried out using a fermentation apparatus (jar fermenter) capable of automatically or semi-automatically controlling the temperature, aeration amount and hydrogen ion concentration.

In addition, when the microorganism used is a microorganism having an alcohol fermentation or lactic acid fermentation ability in addition to the activity of converting curcumins to tetrahydrocurcumins,
During alcohol fermentation or lactic acid fermentation, an O / W type emulsion containing curcumin or a composition containing curcumin or a curcumin or a composition containing curcumin and an O / W type emulsion not containing them are prepared. It is also possible to produce an alcoholic beverage or a lactic acid fermented food containing tetrahydrocurcumins by adding or by adding curcumins or a composition containing curcumins and an oil and an emulsifier or a surfactant. .

The curcumin content in the curcumin-containing composition used to prepare the tetrahydrocurcumin-containing composition according to the present invention is from 0.01% to 99.999%, preferably from 0.1%. It is 99.9%. The total volume of the reaction solution is not limited, but is 0.1 ml to 1000 kl, preferably 1 ml to 1000 kl. The residual amount of the curcumins in the reaction solution after the reaction is 99% to 0.001%, preferably 95% to 0.001% of the curcumins before the reaction. Also,
The amount of tetrahydrocurcumins produced after the reaction is 1 mg to 1 g, preferably 1 mg per 1 g of the curcumins before the reaction.
It is 0 mg to 1 g.

According to the present invention, tetrahydrocurcumins are produced in a composition containing curcumins substantially free of tetrahydrocurcumins, and a composition containing tetrahydrocurcumins can be obtained. Further, according to the present invention, the content of tetrahydrocurcumins in a composition containing both tetrahydrocurcumins and curcumins can be increased to reduce the yellow color of the curcumins in the composition.

According to the present invention, substantially pure tetrahydrocurcumins can be produced. However, the reaction solution is used as it is as a food without separating the bacterial cells of the microorganism used in the reaction, the culture solution or the processed products thereof from the reaction solution. , Alcoholic beverages, food additives, medicines, animal feeds, aquatic feeds, animal medicines, antioxidants, cosmetics and raw materials thereof.

Turmeric is a dry powder of rhizome of a plant belonging to the genus Turmeric, and is a food used as a raw material for turmeric tea and spices. Curcumins purified from turmeric are substances widely used in food production as safe edible natural pigments. Therefore, in the present invention, by using a composition containing curcumin purified from crushed products, extracts, fractions or turmeric of plants belonging to the genus Turmeric as a composition containing curcumin, foods, alcoholic beverages , Food additives, medicines,
An extremely safe composition containing tetrahydrocurcumins, which is suitable for use as animal feed, aquatic feed, veterinary drugs, antioxidants, cosmetics and raw materials thereof, can be obtained.

By the method of the present invention, various compositions containing tetrahydrocurcumins, such as foods, liquors, food additives, medicines, animal feeds, aquatic feeds, animal drugs, antioxidants, cosmetics and their raw materials can be prepared. Can be manufactured. Although the scope of the present invention is not limited by the following production methods, for example, production of turmeric tea containing tetrahydrocurcumin that causes turmeric crushed product and yeast to act, and termeric and yeast to act tetrahydrocurcumin include To produce curry powder, to produce a yeast food containing tetrahydrocurcumins that act on turmeric and yeast, curcumin and yeast containing tetrahydrocurcumins obtained by acting yeast to butter, edible oils such as sesame oil Examples include production of an edible oil containing tetrahydrocurcumins to be dipped and production of an alcoholic beverage containing tetrahydrocurcumins obtained by reacting turmeric and yeast.

Turmeric tea containing tetrahydrocurcumins can be prepared, for example, in the presence of an O / W emulsion.
It can be produced by reacting a turmeric crushed product with a microbial cell, a culture solution or a treated product thereof having an activity of converting a curcumin into a tetrahydrocurcumin.

Curry powder containing tetrahydrocurcumins is, for example, a curcumin of a microbial cell, a culture solution or a processed product thereof which has the activity of converting curcumins to tetrahydrocurcumins in the presence of an O / W type emulsion. Alternatively, by using a composition containing tetrahydrocurcumin obtained by acting on a composition containing turmeric as a raw material of curry powder, the antioxidant function and the pharmacological effect derived from curcumin in curry powder were improved. Curry powder can be produced. That is, the composition obtained in the present invention may be used as a substitute for turmeric in the process of producing curry powder in which various spices are mixed. The yellow color of curry powder is due to curcumin contained in turmeric, which is the raw material of curry powder, so turmeric in curry powder is used for the purpose of producing curry powder with enhanced effects other than coloring and aroma among curcumin's effects. Attempting to increase the amount or amount of curcumins is not practical as it causes more coloring and flavoring than necessary. However, when curcumin or turmeric with an increased content of tetrahydrocurcumins in the present invention is used as a raw material for curry powder, the curry is reduced in color depending on the content of colorless and odorless tetrahydrocurcumins. The functions possessed by tetrahydrocurcumins can be imparted without exceeding the range of the coloring degree and the aroma that are allowed as powder.
However, the tetrahydrocurcumins obtained by the invention may be added to the composition of the curry powder, not as a substitute for turmeric, as long as it is within the edible range.

The yeast food containing tetrahydrocurcumins can be obtained by allowing edible yeast having the activity of converting curcumins to tetrahydrocurcumins to act on a composition containing curcumins or turmeric in the presence of an O / W type emulsion. After the tetrahydrocurcumins are produced by the reaction, the whole reaction product can be collected by freeze-drying or hot-air drying. It is possible to obtain a new yeast food in which the functions of tetrahydrocurcumins are added to the yeast food conventionally produced for nutritional purposes.

The yeast food containing tetrahydrocurcumins can be prepared, for example, in the presence of an O / W emulsion.
After the edible yeast having the activity of converting curcumins to tetrahydrocurcumins is allowed to act on the composition containing curcumins or turmeric to generate tetrahydrocurcumins in the cells, the tetrahydrocurcumins are separated by a method such as centrifugation. After the yeast contained is recovered, it is dried or the like to produce a tetrahydrocurcumin-containing yeast food. Furthermore, the extract obtained by drying or concentrating the yeast containing tetrahydrocurcumin produced in the present invention by hot water extraction or alcohol extraction is tetrahydrocurcumin as compared with the yeast extract conventionally produced for nutritional purposes. It can be used as a new edible yeast extract having a similar function.

The edible oil containing tetrahydrocurcumins is, for example, a microbial cell, a culture solution or a processed product thereof which has the activity of converting curcumins to tetrahydrocurcumins in the presence of an O / W type emulsion. Alternatively, by producing a tetrahydrocurcumin by acting on a composition containing turmeric, mixing with an edible oil, and then recovering the edible oil by a method such as centrifugation to produce an edible oil containing a tetrahydrocurcumin can do. Since the solubility of tetrahydrocurcumins in edible oil is higher than that of curcumins, by using this method, an edible oil containing tetrahydrocurcumins having a low content of curcumins as contaminants can be obtained. Since tetrahydrocurcumins have an antioxidant effect, the obtained edible oil containing tetrahydrocurcumins can be used as a highly stable edible oil that is unlikely to undergo air oxidation, and has various physiological functions of tetrahydrocurcumins. It can be used as edible oil.

Alcoholic beverages containing tetrahydrocurcumins can be prepared, for example, in the presence of an O / W emulsion.
After producing a tetrahydrocurcumin by applying a bacterial cell of a microorganism having an activity of converting curcumin to tetrahydrocurcumin, a culture solution or a treated product thereof to a composition containing curcumin or turmeric to produce tetrahydrocurcumin, a beverage alcohol and Mix and then recover alcohol by a method such as centrifugation, add sweeteners, amino acids, sugars, colorants, flavors as necessary, and add ethanol if it is necessary to further increase the alcohol concentration. Can be manufactured by.

In addition, wine or sake containing tetrahydrocurcumins is prepared by fermenting grapes or rice with wine yeast or sake yeast having the activity of converting curcumins into tetrahydrocurcumins, and during or after fermentation. For example, it can be produced by adding curcumin or turmeric in the presence of an O / W type emulsion to generate tetrahydrocurcumin and then squeezing. Although the anti-atherogenic effect derived from the antioxidant activity of red pigments and tannins contained in red wine has been attracting attention, the present invention has enhanced the antioxidant activity without affecting the color or taste of white wine or Sake can be produced.

Fermented dairy products containing tetrahydrocurcumins are fermented milk using lactic acid bacteria having the activity of converting curcumins to tetrahydrocurcumins, and are fermented during or after fermentation, for example, O / W type. It can be produced by adding curcumins or turmeric in the presence of an emulsion.

As the microorganism used in the present invention, any microorganism can be used as long as it has an activity of converting curcumins into tetrahydrocurcumins. Examples of microorganisms include bacteria, actinomycetes, yeasts, filamentous fungi, mushrooms,
Examples include algae.

[0065] Examples of the bacteria, for example Aserobakuta genus (Acetobacter), Achromobacter sp (Achromobacte
r ), genus Arthrobacter , genus Bacillus , genus Bifidobact
erium), the genus Brevibacterium (Brevibacterium),
Cellulomonas sp (Cellulomonas), the genus Chromobacterium (Chromobacterium), Shitobakuta genus (Citobacte
r), Kurosutoridjiumu genus (Clostridium), Corynebacterium (Corynebacterium), Enterococcus (Enterococcus), Euinia genus (Erwinia), the genus Escherichia (Escherichia), Flavobacterium (Flavobacterium), the genus Gluconobacter (Gluconob
acter ), Halobacterium ( Halobacterium ), Klebsiella ( Klebsiella ), Leuconostoc ( Le
uconostoc ), genus Micrococcus , genus Pediococcus , genus Propionibacterium , genus Protaminobacter , genus Providencia
a), the genus Pseudomonas (Psudomonas), Serratia (Serratia), Streptomyces genus (Streptobac
terium ), Streptococcus genus ( Streptococcus ),
The genus Xanthomonas (Xanthomonas), Zymomonas genus (Zy
momonas ), Lactobacillus ( Lactobacillus ), Pediococcus ( Pediococcus ), Staphylococcus ( Staphylococcus ), and the like.

The actinomycetes include, for example, Actinoplanes and Micromonospora.
pora), Streptomyces (Streptomyces), and the like. Examples of yeasts include, for example, the genus Deballomyces ( De
baryomyces ), Saccharomyces ( Saccharomyces
), Pichia k , Kluyveromyces ( K
luyveromyces ), Torulaspora ( Torulaspora ),
Kyandjida genera (Candida), Shubia genus (Ashbya), breccias Tano My genus (Brettanomyces), Cryptococcus (Cryptococcus), eremotherium genus (Eremothe
cium ), genus Issatchenkia (genus Issatchenkia ), genus Klockera (genus Klockera ), genus Lipomyces ( lipomyces ),
Meth Shuni Kou Oia genus (Metschnikowia), Rodotorua genera (Rhodotorula), Schizosaccharomyces genus (Shiz
osaccharomyces ), Zygosacc
haromyces ) and the like.

Examples of filamentous fungi include the genus Acremonium and the genus Actinomucor.
), Aspergillus (Aspergillus), Out Leo Basi an indium belonging to the genus (Aureobasidium), Bakusera genus (Backus
ella), Botrytis genus (Botrytis), Charara genus (Chal
ara ), Claviceps ( Claviceps ), Corticium ( Corticium ), Cryphonectria ( Cryphonect)
ria), Yuurochiumu genus (Eurotium), Fusarium (Fusarium), Geotorichumu genus (Geotrichum), Monascus genus (Monascus), Mortierella genus (Mortierella
), Mucor (Mucor), Miroteshiumu genus (Myrothe
cium ), Neurospora ( Neurospora ), Paecilomyces ( Paecilomyces ), Penicillium ( Peni )
cilium ), Pestalotiopsis ( Pestalotiopsis ), Rhizomucor ( Rhizomucor ), Rhizopu ( Rhizopu )
s ), the genus Sclerotinia , the genus Syncephalastrum , and the genus Trichoderma .

Mushrooms include, for example, the genus Agaricus ( Ag
aricus), Aguroshibe genus (Agrocybe), Arumiraria genus (Armillaria), Aurikuraria genus (Auricularia),
Furamurina genus (Flammulina), Ganoderma sp (Ganoderm
a), Gurifora genus (Grifola), Hipushijigasu genus (Hypsizigus), Irupekkusu genus (Irpex), Renchinura genus (Lentinula), Repisuta genus (Lepista), Riofiriumu genus (Lyophyllum), Maiko streptomycin de Neu death genus (Mycoleptodonoides), Naematoroma Genus ( Naematol
oma ), genus Panellus , genus Polita ( Pholio )
ta ), Pleurotus ( Pleurotus ), Pycnoporus ( Pycnoporus ), Tremella ( Tremella ), Trichoroma ( Tricholoma ), Bolvariella ( Volvariell )
a ) etc.

As the algae, for example, genus Analipus ( Anar)
ipus), Chondrus genus (Chondrus), Eisenia species (Ei
senia), Yushuma species (Eucheuma), Furuseraria genus (Furcellaria), Jigachina genus (Gigartina), Hijikia genus (Hizikia), click Gela Maniera genus (Kjellama
niella), Laminaria species (Laminaria), macro Chris Chis genus (Macrocrystis), Petaronia genus (Petalonia
), Porphyra genus (Porphyra), Rodjimenisu genus (Rhody
menis ), genus Scytosiphon ( Scytosiphon ), genus Spirulina ( Spirulina ), genus Undaria ( Undaria ) and the like.

Specifically, yeasts such as Devalyomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora and Candida, lactic acid bacteria such as Lactobacillus and Pediococcus, and grapes such as Staphylococcus and the like Examples of microorganisms include cocci. More specifically suitable strains include, for example, Devalyomyces
Debaryomyces hansenii ATCC-20261, Debaryomyces hanseni
i ) IFO-0094, Saccharomyces cerevisiae ( Sacch
aromyces cerevisiae ) Dry yeast Lallmand, Saccharomyces cerevisiae
e ) IAM-4500, Saccharomyces cerevisiae ( Sacch
aromyces cerevisiae ) IAM-4519, Saccharomyces cerevisiae ATCC-77
54, Saccharomyces
cerevisiae ) FERM-P-6189, Saccharomyces cerevisiae IFO-2044, Saccharomyces cerevisiae
ae ) ATCC-20018, Saccharomyces cerevisiae FERM-P-6213, Saccharomyces cerevisiae
FERM-P-6214, Saccharomyces cerevisiae ( Sacc
haromyces cerevisiae ) FERM-P-7614, Saccharomyces cerevisiae FERM-
P-7615, Pichiasubpelli
culosa) ATCC-16766, Pichia anomala (Pichia an
omala) ATCC-2149, Pichia Men bra seedlings Faye tumefaciens (Pichia membranaefaciens) IAM-4986, Pichia Men bra seedlings Faye tumefaciens (Pichia membranaefacien
s ) ATCC-36908, Pichia kluyveri
) ATCC-9768, Kluyveromyces marxianus (IFO-0433), Kluyveromyces marxianus ( Kluyveromyces marxianus )
IFO-1090, Kluyveromyces Polysporus ( Kluyve
romyces polysporus) ATCC-22028, Torulaspora-Deruburukkii (Torulaspora delbrueckii) IAM-4816, Kyandjida-Uchirisu (Candida utilis) ATCC-9950, Kyandjida-Famata (Candida famata) ATCC-2560,
Lactobacillus planta
rum ) LPT (Kyowa High Foods Co., Ltd.), Staphylococcus carnosus M72 (TE
XEL), Pediococcus acidophilus ( Pedioc
occus acidilactici ) P2M120 (made by TEXEL), Staphylo coccus xylosus 2)
P2M120 (made by TEXEL) is an example.

Any of these strains can be used alone or in combination. Also, curcumins can be converted to tetrahydrocurcumins even in a mutant strain which is mutated by an artificial mutagenesis method such as ultraviolet irradiation, X-ray irradiation, treatment with a mutagenic agent, or genetic manipulation, or a naturally mutated mutant strain. Any competent strain can be used in the present invention.

For the culture of these microorganisms, a natural medium containing a carbon source, a nitrogen source and other nutrients, a semi-synthetic medium, a synthetic medium can be used as long as the medium can grow in the medium used for culturing ordinary yeasts, bacteria and the like. Any medium such as a medium can be used. Examples of the carbon source include starch, dextrin, sucrose, glucose, mannose, fructose, raffinose, rhamnose, inositol, lactose, xylose, arabinose, mannitol and molasses, and these can be used alone or in combination. Furthermore, hydrocarbons, alcohols, organic acids and the like may be used depending on the assimilation ability of the bacterium.

Examples of the nitrogen source include ammonium chloride, ammonium nitrate, ammonium sulfate, sodium nitrate, urea, peptone, meat extract, yeast extract, dry yeast, corn steep liquor, soybean powder, casamino acid, etc., either alone or in combination. Can be used. As inorganic salts, sodium chloride, potassium chloride, magnesium sulfate, calcium carbonate, potassium dihydrogen phosphate,
Examples thereof include ferrous sulfate, calcium chloride, manganese sulfate, zinc sulfate and copper sulfate, which can be used alone or in combination.

Examples of the trace component include vitamins such as biotin, thiamine or nicotinic acid, amino acids such as β-alanine or glutamic acid, and these can be used alone or in combination. As the culturing method, a liquid culturing method, particularly a deep agitation culturing method is suitable. Culture temperature 1
It is carried out at 0 to 80 ° C., preferably 10 to 60 ° C., particularly preferably 20 to 40 ° C., pH 2 to 11, pH 3 to 10, preferably pH 5 to 8, and usually 1 to 7 days. P of medium
Ammonia water, ammonium carbonate solution, or the like is used for H adjustment.

As the processed product of the culture solution used in the present invention,
Concentrate, dried product, frozen product of a culture solution of a microorganism having an activity of converting curcumins to tetrahydrocurcumins,
Refrigerated, freeze-dried, heated, pressurized, ultrasonically crushed,
Examples include surfactant or organic solvent-treated products, lytic enzyme-treated products, and the like. In addition, as the treated microbial cells, dried, frozen, chilled, freeze-dried, lyophilized, heated, pressurized, and ultrasonically crushed products of microbial cells having the activity of converting curcumins to tetrahydrocurcumins , Surfactants or organic solvent-treated products, lytic enzyme-treated products, immobilized cells or enzymes purified from cells.

For purification of the enzyme from the bacterial cells, a general protein purification method can be used. For example, homogenizer, glass beads, ammonia dissolution, disruption of bacterial cells by enzyme method, filtration, recovery of enzyme solution by centrifugation, etc., salting out, organic solvent precipitation, enzyme recovery by antibody, concentration by dialysis, etc. External filtration, gel filtration, electrophoresis, separation by liquid phase partitioning method, chromatography using an ion exchanger, adsorbent, affinity adsorbent, etc., batch method, crystallization, etc. can be used alone or in combination. .

Curcumins can be obtained as reagents, for example, from Sigma-Aldrich Japan. The composition containing curcumin is not particularly limited, and examples thereof include a composition obtained by treating a plant belonging to the genus Turmeric. The plants belonging to the genus Turmeric include, for example, rhubarb (commonly called spring turmeric), turmeric (generally,
Called autumn turmeric. ), Plants such as zedoary. The treatment method is not particularly limited, and examples thereof include pulverization, drying and extraction. The curcumin-containing composition also includes general food products such as curry powder and turmeric tea.

Examples of the composition containing curcumin include a composition obtained by adding curcumin or a composition containing curcumin to food, liquor, feed, cosmetics which does not contain curcumin. Examples of curcumin-free foods, alcoholic beverages, feeds, cosmetics include:
Examples thereof include alcohol for beverages, edible oil and fruit juice. Curcumins and compositions containing curcumins can be easily obtained as commercial products as foods.

The present invention will be described below with reference to examples.
This example does not limit the present invention.

[0080]

EXAMPLES Example 1 Conversion Reaction in the Presence of O / W Emulsion 1) Preparation of O / W Emulsion 63.6 g of glycerin (manufactured by Kishida Chemical Co., Ltd.), polyglycerin monooleate (Poem J-0381, RIKEN VITAMIN) (Manufactured by the company), 5 g of enzymatically decomposed lecithin (Elmizer AC, manufactured by Trurecitin Industry Co., Ltd.), and 20 g of pure water are mixed in a homogenizer cup, and a homogenizer (AM
-7 type, manufactured by NISSEI) and stirred almost uniformly. MCT (Panasate 810, NOF Corporation)
10 g, and stirred at 17,000 rpm for 10 minutes,
An O / W type emulsion containing no curcumin was prepared. The average particle size of this O / W emulsion is 110 n.
m (particle size measurement: model N4MD, manufactured by COULTER).

2) Conversion reaction by cells in the presence of O / W type emulsion Debaryomyces hansenii (PRISCA, LACTO LABO) YM
Cultured in a medium (manufactured by Nissui) at 25 ° C. for 2 days and night (0.
(03 g cells / ml) and 2.0 ml Eppendorf tube were centrifuged at 12,000 rpm for 10 minutes to obtain cells. The cells were suspended in the above-described curcumin-free O / W type emulsion to 0.4 g cells / ml to make 1 ml. 2 mg of curcumin powder (manufactured by Sigma-Aldrich Japan) was added to the suspension, and the mixture was vigorously shaken at 200 rpm using a shaking culture machine (TC-C-500R type, manufactured by Takasaki Scientific Instruments Co., Ltd.) at 37 ° C. for 48 hours. I kept it warm.

Tetrahydrocurcumin in the reaction solution was analyzed under the following HPLC measurement conditions. HPLC conditions: Solvent: 50% acetonitrile aqueous solution containing 0.05% trifluoroacetic acid Flow rate: 1 ml / min, Detection: 280 nm As a result, after 48 hours, 0.66 mg / ml of tetrahydrocurcumin was produced.

Example 2 1) Preparation of Oil Containing Curcumin 2 g of curcumin (manufactured by Sigma-Aldrich Japan) was added to 10 g of MCT (Panasate 810, manufactured by NOF CORPORATION), oil bath (BO-11 type, Yam
(produced by ato) was heated at 115 ° C. for 10 minutes to prepare a curcumin-containing oil.

2) Preparation of Curcumin-Containing O / W Emulsion (G) 63.6 g of glycerin, 5 g of polyglycerin monooleate (Poem J-0381, manufactured by Riken Vitamin Co.), enzymatically decomposed lecithin (Elmizer AC, Trulecithin industrial stock) (Manufactured by the company) 1 g, 20 g of water are mixed in a homogenizer cup, and a homogenizer (AM-7 type, NIS)
The mixture was stirred almost uniformly with SEI). The entire amount of the curcumin-containing oil was added to this, and the mixture was stirred at 17,000 rpm for 10 minutes to produce an O / W type emulsified curcumin.
Curcumin concentration in the O / W emulsion is 2.5 m
g / ml, average particle size 150 nm (particle size measurement:
Model N4MD, manufactured by COULTER). Book O
The / W emulsion is hereinafter simply referred to as O / W emulsion (G).

3) Preparation of O / W type emulsion (W) containing curcumin 5 g of polyglycerin monooleate (Poem J-0381, manufactured by Riken Vitamin Co.), 1 g of enzymatically decomposed lecithin (Elmizer AC, manufactured by Trulecithin Industry Co., Ltd.), Water 8
3.6 g was mixed in a homogenizer cup, and stirred with a homogenizer (AM-7 type, manufactured by NISSEI) almost uniformly. Add all of the curcumin-containing oil to this and add 1
The mixture was stirred at 7,000 rpm for 10 minutes to prepare an O / W type emulsified curcumin. The curcumin concentration in the O / W type emulsion was 2.5 mg / ml, and the average particle size was 150 nm (particle size measurement: model N4MD, COUL
TER). Hereinafter, the present O / W type emulsion is simply referred to as an O / W type emulsion (W).

4) Debaryomyces hansenii (PRISCA, manufactured by LACTO LABO) was used as a Y-type conversion reaction in the presence of an O / W type emulsion.
The cells were cultured in M medium (manufactured by Nissui Co., Ltd.) at 25 ° C. for 2 days and night (0.03 g bacterial cells / ml), and centrifuged in a 2.0 ml Eppendorf tube at 12,000 rpm for 10 minutes to obtain bacterial cells. 200 μ of O / W type emulsion curcumin stock solution (100%) or O / W type emulsion curcumin water dilution solution (25 and 50%)
0.4 g of cells / ml was suspended in 1 to give a reaction solution, which was then shaken at 30 ° C. for 4 hours in a shaking culture machine (TC-C-
200 rpm using 500R type, manufactured by Takasaki Scientific Instruments Co., Ltd.
I kept it warm while shaking vigorously. After 4 hours, add 800 μl of ethanol and vortex to 12000 rp.
The supernatant obtained by centrifugation for 1 min at
Analysis was performed to calculate the concentration of curcumin and tetrahydrocurcumin in the reaction solution.

Tetrahydrocurcumin in the reaction solution was analyzed under the above-mentioned HPLC analysis conditions. As a result, 25%,
0.047 mg / ml to 0.275 mg / m 2 when adding O / W emulsion at 50% and 100% volume
l of tetrahydrocurcumin was produced. The results are shown in Table 1.

Comparative Example 1 Debaryomyces hansenii (PRISCA, manufactured by LACTO LABO) was used as Y
The cells were cultured in M medium (manufactured by Nissui Co., Ltd.) at 25 ° C. for 2 days and night (0.03 g bacterial cells / ml), and centrifuged in a 2.0 ml Eppendorf tube at 12,000 rpm for 10 minutes to obtain bacterial cells. The cells were suspended in 200 μl of water so that the concentration was 0.4 g of cells / ml to give a reaction solution, and 4 m
1/20 volume of a g / ml curcumin ethanol solution was added, and this was shaken for 4 hours at 30 ° C (TC-C-
200 rpm using 500R type, manufactured by Takasaki Scientific Instruments Co., Ltd.
I kept it warm while shaking vigorously. The ethanol concentration in the reaction solution is 5%. After 4 hours, 800 μl of ethanol
The supernatant obtained by adding and vortexing and centrifuging at 12,000 rpm for 1 minute was subjected to HPLC analysis under the following conditions to calculate the curcumin and tetrahydrocurcumin concentrations in the reaction solution. The HPLC analysis conditions are the same as in Example 1.
The results are shown in Table 1.

[0089]

[Table 1]

When O / W type emulsified curcumin is used, THU1 can be efficiently produced.

Example 3 Microbial conversion reaction using O / W type emulsified curcumin The preparation method and concentration were the same as in Example 1. 10 cells were prepared by suspending the cells in 200 μl of an O / W type emulsified curcumin stock solution (cell concentration of 0.4 g cells / ml) and incubated at 30 ° C. with vigorous shaking.
After 4, 8, 24 and 72 hours, 2 bottles of ethanol each, 80
The reaction was stopped by adding 0 μl, and the supernatant obtained by centrifugation at 12,000 rpm for 1 minute was subjected to HPLC analysis.

In the reaction with the initial U1 concentration of 2.5 mg / ml using the O / W type emulsion (G), the production amount of THU1 increased up to 72 hours, which was 10 times or more than that of the ethanol solution added. It became 0.3 mg / ml. The results are shown in Table 2.

Comparative Example 2 Cells were suspended in water (0.4 g cells / ml cell concentration), and 1/10 volume of a 18 mg / ml curcumin ethanol suspension was added (curcumin final concentration 1 .8m
(g / ml) was kept warm at 30 ° C. with vigorous shaking.
After 4, 8, 24 and 72 hours, 2 bottles of ethanol each, 80
The reaction was stopped by adding 0 μl, and the supernatant obtained by centrifugation at 12,000 rpm for 1 minute was subjected to HPLC analysis.
The results are shown in Table 2.

[0094]

[Table 2]

As shown in Table 2, most of U1 remained unconverted by the conventional ethanol method. As shown in Table 2, in the conversion reaction using the O / W type emulsified curcumin, 9% of the curcumin added 72 hours after the reaction was added.
In the conversion reaction using curcumin dissolved in ethanol, 0% or more could be converted to tetrahydrocurcumin, while 10% or less of the curcumin added 72 hours after the reaction could be converted to tetrahydrocurcumin.

Example 4 Application of O / W Emulsified Curcumin to Enzymatic Reaction PR according to Biomedical Series 10 "Gene Experiment Method by Yeast" (edited by Masayuki Yamamoto, Yodosha)
An ISCA cell disruption solution (crude enzyme solution) was prepared. That is, PR
25 ml (0.045 g bacterial cells / ml) of ICSCA cultured in YM medium was centrifuged (6,000 rpm, 20 minutes), and the obtained precipitate was suspended in 2 ml of ice-cooled DIW and transferred to an Eppendorf tube. After vortexing and washing, 1.1 g ml of bacterial cells was obtained by centrifugation again.
To the cells, 1.2 ml of Lysis buffer (the following composition) and 1.6 ml of glass beads were added and cooled on ice. This is a vortex mixer (MULTI BEADS SHOCKER MB-200,
Vigorously stirred with Yasui Kikai Co., Ltd.). The operation of stirring for 30 seconds and then cooling on ice for 1 minute was repeated 10 times. After this, centrifuge (15,000 rpm, 30 minutes, 4 ° C)
The resulting supernatant was recovered and used as a crude enzyme solution.

Lysis buffer composition (concentration is final concentration): 20
mmol / L Tris-HCl (pH 7.5), 1 m
mol / L EDTA, 5 mmol / L MgCl2,
50 mmol / L KCl, 5% glycerol, 3 m
mol / L dithiothreitol, 1 mmol / L P
MSF (The above is autoclaved at 120 ° C. for 20 minutes and stored. Dithiothreitol and PMS are added just before use).

0.5 mol / L per 100 μl of the crude enzyme solution
ATP, 100 mmol / L NADH, 100 mm
1/100 volume of ol / L NADPH and O / W type emulsified curcumin (G) prepared in Example 2
Or use 1 of O / W type emulsified curcumin (W)
/ 10 volume was added, and the mixture was kept warm at 37 ° C with shaking. 0.5,
After 1, 2 and 24 hours, it was diluted with 10 volumes of ethanol and the centrifuged supernatant was subjected to HPLC. The results are shown in Table 3.

[0099]

[Table 3]

By using the O / W emulsified curcumin, the curcumin was almost completely converted to tetrahydrocurcumin after 24 hours.

Comparative Example 3 Debaryomyces hansenii (PRISCA, LACTO LABO) was treated with YM.
Cultured in a medium (manufactured by Nissui) at 25 ° C. for 2 days and night (0.
(03 g / ml) and a 2.0 ml Eppendorf tube were centrifuged at 12,000 rpm for 10 minutes to obtain bacterial cells. The cells were suspended in 200 μl of water to a concentration of 0.4 g / ml to prepare a reaction solution, and 2 mg / ml thereof was added thereto so that the concentration of curcumin in the reaction solution was 0.1 mg / ml.
1/20 volume of the curcumin ethanol solution of ml (final ethanol concentration 5%) or 1/10 volume of the curcumin ethanol solution of 1 mg / ml (final ethanol concentration 1
0%) or 0.5 mg / ml of curcumin ethanol solution was added to 1/5 volume (final ethanol concentration: 20%).
%), And this was kept at 30 ° C. for 4 hours while vigorously shaking at 200 rpm using a shaking culture machine (Takasaki Scientific Instruments, TC-C-500R type). After 4 hours, add 8
Add 00 μl and vortex, 12,000 rpm
The supernatant obtained by centrifuging for 1 minute was analyzed by HPLC under the following conditions to calculate the curcumin and tetrahydrocurcumin concentrations (THU1 concentration) in the reaction solution. The HPLC analysis conditions are the same as in Example 1. The results are shown in Table 4.

[0102]

[Table 4]

When the ethanol concentration was 5% and 10%, T
The amount of HU1 produced was 70 μg / ml, while 20
%, The amount of THU1 produced was as low as 15 μg / ml, suggesting that the enzyme in the reaction system may have been inactivated by high-concentration ethanol.

Comparative Example 4 Debaryomyces hansenii (PRISCA, LACTO LABO) was treated with YM.
Cultured in a medium (manufactured by Nissui) at 25 ° C. for 2 days and night (0.
(03 g cells / ml) and 2.0 ml Eppendorf tube were centrifuged at 12,000 rpm for 10 minutes to obtain cells. 20 μl of the cells were added to 0.4 g cells / ml
To obtain a reaction solution, and add 2 mg / m
l, or 4 mg / ml, or 6 mg / ml, 8
A 1/20 volume of a curcumin ethanol solution of mg / ml or 10 mg / ml was added so that the ethanol concentration in the reaction solution would be 5%, and this was added at 30 ° C. for 4 hours.
Using a shaking culture machine (TC-C-500R type, manufactured by Takasaki Kagaku Kikai Co., Ltd.), the temperature was kept violently shaking at 200 rpm. After 4 hours, 800 μl of ethanol was added, the mixture was shaken with a vortex and centrifuged at 12,000 rpm for 1 minute, and the obtained supernatant was analyzed by HPLC under the following conditions to calculate the curcumin and tetrahydrocurcumin concentrations in the reaction solution. . The HPLC analysis conditions are the same as in Example 1. The results are shown in Table 5.

[0105]

[Table 5]

Undissolved curcumin was observed in those to which a curcumin ethanol solution having a concentration higher than 4 mg / ml was added. Produced tetrahydrocurcumin (THU1)
The concentration showed a maximum value of 120 μg / ml for U1 of 4 mg / ml and 6 mg / ml, and even if the concentration of added curcumin was increased, the concentration of THU1 produced was not increased.

[0107]

INDUSTRIAL APPLICABILITY According to the present invention, a method for industrially producing tetrahydrocurcumins using a microbial cell, a culture, or a treated product thereof, without depending on the method of chemically-synthesizing hydrogenation using a catalyst, and A method for producing a composition containing tetrahydrocurcumins can be provided.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) (C12P 7/26 C12R 1:72 C12R 1:44) 1:84 (C12P 7/26 C12R 1:85 C12R (1: 645) (C12P 7/26 C12R 1:72) (C12P 7/26 C12R 1:84) (C12P 7/26 C12R 1:85)

Claims (9)

[Claims] 1. A compound of formula (I) in the presence of an O / W emulsion. [Wherein R ¹ , R ² , R ³ and R ⁴ in the formula (I) are the same or different and each represents hydrogen, hydroxy or lower alkoxy], or a curcumin or curcumin derivative (hereinafter, simply referred to as curcumin) Formula (II): [In the formula (II), R ¹ , R ² , R ³ and R ⁴ are the same or different and each represents hydrogen, hydroxy or lower alkoxy] or a tetrahydrocurcumin or tetrahydrocurcumin derivative (hereinafter, simply referred to as tetrahydrocurcumins A tetrahydrocurcumin or a tetrahydrocurcumin characterized by allowing a bacterial cell, a culture solution or a treated product thereof having an activity of converting into a curcumin or a composition containing the curcumin to act on the composition. A method for producing a composition. 2. The production method according to claim 1, wherein the curcumin or the composition containing the curcumin is contained in an O / W type emulsion. 3. The production method according to claim 1, wherein the O / W emulsion is obtained by mixing an oil, an emulsifier or a surfactant and an aqueous medium. 4. The average particle diameter of the O / W emulsion is
The method according to claim 1, which has a thickness of 10 nm to 10 μm. 5. The method according to claim 1, wherein the composition containing curcumin is a composition obtained by treating a plant belonging to the genus Turmeric. 6. The microorganism is a microorganism belonging to the genera Devalyomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, Lactobacillus, Staphylococcus, or Pediococcus. The method described in any one of. 7. A fungus body, a culture solution or a treated product of a microorganism having an activity of converting curcumins to tetrahydrocurcumins in the presence of oils and an emulsifier or a surfactant in an aqueous medium, is treated with curcumins or A method for producing a tetrahydrocurcumin or a composition containing a tetrahydrocurcumin, which comprises acting on a composition containing a curcumin. 8. The method according to claim 7, wherein the composition containing curcumin is a composition obtained by treating a plant belonging to the genus Turmeric. 9. The microorganism according to claim 7, wherein the microorganism belongs to the genus Devalyomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, Lactobacillus, Staphylococcus, or Pediococcus. The method described in.