JPH11235192A - Method for producing tetrahydrocurcumin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a highly pure tetrahydrocurcumin having excellent antioxidative properties and useful for foods, alcoholics, medicaments, cosmetics, etc., in a high yield, by bringing fungus body of a specific microorganism, etc., specific curcumin derivative. SOLUTION: This production method of tetrahydrocurcumins is to bring fungus body of a microorganism having an activity to transform a derivative expressed by formula I (R<1> to R<4> are each H, hydroxy or a lower alkoxy) to a tetrahydrocurcumin derivative expressed by formula II, their culture solution or its treated product, to act on a curcumin derivative of formula I or a composition containing the same and obtain the tetrahydrocurcumin of formula II. The microorganism preferably belongs to the genus Debaryomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, Lactobacillus, Staphylococcus or Pediococcus.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for producing tetrahydrocurcumin and a composition containing tetrahydrocurcumin which are useful in the fields of food, alcohol, medicine, animal feed, marine feed, animal medicine, and cosmetics.

[0002]

2. Description of the Related Art Curcumin is well known as a yellow pigment contained in plants belonging to the genus Turmeric of the tropical ginger family. Turmeric rhizome dry powder (turmeric) and its purified product, curcumin, are used as flavoring agents and coloring agents for foods. Curcumin is known to have a pharmacological effect such as an antioxidant action, an anti-inflammatory action, a cholesterol-reducing action, a carcinogenesis-suppressing action, and is used as a safe medicinal ingredient derived from food.

It is known that tetrahydrocurcumin has a stronger antioxidant activity than curcumin [JP-A-2-49747, JP-A-2-51595.
Gazette, Bioscience Biotechnology and Biochemistry (Biosci. Biotech. Biochem.),
59 , 1609 (1995)]. In addition, since tetrahydrocurcumin is colorless and odorless, it is a material that overcomes the disadvantages of curcumin, particularly when coloring is an adverse effect.

As a method for producing tetrahydrocurcumin, there is known a method for producing curcumin by hydrogenation using a metal catalyst. However, in Japan, synthetic tetrahydrocurcumin produced by this method cannot be used for food and drink applications. It is known that a part of curcumin administered by intravenous injection is reduced and converted to tetrahydrocurcumin [Xenobiotica (Xenobiotica), 8, 761 (1978)]. This suggests that a mechanism for converting curcumin to tetrahydrocurcumin exists in vivo. However, the details of the mechanism have not been clarified to date, and no reductase has been identified. Furthermore, it is not known that microorganisms have such activity at all. For these reasons, there has been no report on a technique for producing tetrahydrocurcumin on an industrial scale by adding hydrogen to curcumin by the action of an organism, including microorganisms, or an enzyme.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a process for producing industrial tetrahydrocurcumins using microbial cells, cultures or processed products thereof, irrespective of the method of chemical synthesis of hydrogenation using a catalyst. And a method for producing a composition containing tetrahydrocurcumins. Another object of the present invention is to provide a microorganism containing tetrahydrocurcumin.

[0006]

According to the present invention, there is provided a compound of the formula (I)

[0007]

Embedded image

[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]] (hereinafter simply referred to as curcumins) of formula (II)

[0009]

Embedded image

[In the formula (II), R ¹ , R ² , R ³ and R
⁴ are the same or different and represent hydrogen, hydroxy or lower alkoxy]], a microbial cell, a culture solution or a treated product thereof, having an activity of converting into a tetrahydrocurcumin derivative (hereinafter, simply referred to as tetrahydrocurcumins) Is reacted with curcumin or a composition containing curcumin to produce tetrahydrocurcumin, and the produced tetrahydrocurcumin is collected.A method for producing tetrahydrocurcumin, wherein curcumin is converted to tetrahydrocurcumin A method for producing a composition containing tetrahydrocurcumins, which comprises reacting microbial cells, culture broth, or a processed product thereof with a curcumin-containing composition to produce tetrahydrocurcumins. Containing no curcumin and curcumin Curcumin or a composition containing curcumin is added to the resulting product, and cells of a microorganism having an activity of converting curcumin into tetrahydrocurcumin, a culture solution, or a processed product thereof are allowed to act on the resultant to produce tetrahydrocurcumin. The present invention relates to a method for producing a composition containing tetrahydrocurcumins, which is characterized by being produced.

[0011] The present invention also provides a method for producing a microorganism containing tetrahydrocurcumins, which comprises culturing a microorganism having an activity of converting curcumins into tetrahydrocurcumins in a medium containing curcumins. The present invention relates to microorganisms containing the same. Furthermore, the present invention provides a method for producing a tetrahydrocurcumin-containing edible oil, which comprises immersing a microorganism containing tetrahydrocurcumin or a processed product thereof in edible oil, and converting the microorganism containing tetrahydrocurcumin or a processed product thereof into liquor. The present invention relates to a method for producing a liquor containing tetrahydrocurcumin, which is characterized by being immersed.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION In formulas (I) and (II), the alkyl portion of lower alkoxy represented by R ¹ , R ² , R ³ and R ⁴ is a straight-chain or branched alkyl having 1 to 6 carbon atoms. For example, methyl, ethyl, propyl, isopropyl,
Butyl, sec-butyl, tert-butyl, pentyl, hexyl and the like. 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 that is hydroxy is preferable.

Specific curcumins include U1 [diferuloylmethane (E, E) -1,
7-bis (4-hydroxy-3-methoxyphenyl) -1,6-heptadiene-3,
5-dione, sometimes simply curcumin], U
2 [demethoxycurcumin,
bis (4-hydroxy-3-methoxycinnamoyl) methane], U3
[Bisdemethoxycurcumi
n), 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-dihydr
oxyphenyl) -1,6-heptadiene-3,5-dione], U1, U2, U3 and DHU1 are preferably used, and U1 and DHU1 are particularly preferably used.

A reaction for producing tetrahydrocurcumin by reacting a microbial cell, a culture solution or a treated product thereof with a curcumin or a composition containing curcumin having the activity of converting curcumin into tetrahydrocurcumin State. A culture solution of the microorganism, cells or a treated product thereof and curcumin or a composition containing curcumin are added to an aqueous medium such as water to form a reaction solution, and the reaction solution is 0 to 100 ° C, preferably 10 to 100 ° C. 60
C., particularly preferably 22-45 ° C., pH 2-11, preferably 3-9, particularly preferably 4-8,
The reaction is carried out for 0.01 to 168 hours, preferably for 0.5 to 72 hours.

The amount of the culture solution, microbial cells, or their treated products of the microorganisms to be added to the reaction solution is not particularly limited as long as it has the activity of converting curcumins into tetrahydrocurcumins. 1 μg to 80
A reaction solution is prepared so as to have a concentration of 0 mg / ml, preferably 10 to 500 mg / ml. Curcumin or a composition containing curcumin is contained in the reaction solution as curcumin in an amount of 0.001 to 500 mg / ml, preferably 0.01 to 500 mg / ml.
Add to よ う 100 mg / ml.

In the reaction solution, a buffer, a surfactant, an organic solvent, a coenzyme, and the like can be present as necessary. Examples of the buffer include sodium hydrogen phosphate, glycine, N-2-hydroxyethylpiperazine-N′-
2-ethanesulfonic acid, ammonium acetate, tris (hydroxymethyl) methylglycine and the like can be mentioned.

Examples of the surfactant include Triton X100. As an organic solvent, for example, methanol,
Examples include ethanol, isopropanol, butanol, propylene glycol, butylene glycol, glycerin, acetone and the like. Examples of the coenzyme include beta nicotinamide adenine dinucleotide, beta nicotinamide adenine dinucleotide phosphate, reduced beta nicotinamide adenine dinucleotide, reduced beta nicotinamide adenine dinucleotide phosphate, and the like.

The extraction of tetrahydrocurcumins formed in an aqueous medium is carried out by means of industrial extraction and purification of compounds such as organic solvent extraction, centrifugation, column chromatography, freeze-drying, recrystallization and hot-air drying. Extraction and purification can be carried out by a method generally used in the above. Analysis and quantification of curcumins and tetrahydrocurcumins in the fermentation reaction can be performed by high performance liquid chromatography, and structure determination can be performed by mass spectrometry and nuclear magnetic resonance.

When microbial cells are used, curcumin or a composition containing curcumins and microbial cells are:
It is not necessary to mix them uniformly in the reaction tank, and they may be isolated from each other by a membrane that allows the permeation of curcumins but does not allow the passage of microbial cells. Alternatively, the microbial cells may be embedded in an alginate gel, an agarose gel, or the like, and then added to a reaction vessel to obtain a tetrahydrocurcumin or a composition containing the tetrahydrocurcumin. Alternatively, a tetrahydrocurcumin or a composition containing tetrahydrocurcumin can also be obtained by reacting microbial cells immobilized in a hollow membrane thread (hollow fiber) with curcumin or a composition containing curcumin. . These techniques are effective when it is desired to avoid mixing microbial cells into a target tetrahydrocurcumin or a composition containing tetrahydrocurcumin, or when it is desired to recover only microbial cells.

For the reaction between the curcumin or the composition containing the curcumin and the microbial cells, a fermenter (jar fermenter) capable of automatically or semi-automatically controlling the temperature, aeration rate and hydrogen ion concentration is used. It may be used, for example, by simply mixing a microorganism and a composition containing curcumin or curcumin in a wooden barrel without controlling the temperature, aeration rate, or hydrogen ion concentration, and then allowing the mixture to stand for a certain period of time. A simple method may be used.

When the microorganism used is a microorganism having alcohol fermentation or lactic acid fermentation ability in addition to the activity of converting curcumins into tetrahydrocurcumins,
By adding curcumins or a composition containing curcumins during alcohol fermentation or lactic acid fermentation, an alcoholic beverage or lactic acid-fermented food containing tetrahydrocurcumins can also be produced.

The curcumin content in the curcumin-containing composition used for producing the tetrahydrocurcumin-containing composition according to the present invention is from 0.01% to 99.9%.
It is 99%, preferably 0.1% to 99.9%. The total volume of the reaction product of the composition containing the microorganism and the curcumins is not limited, but may be from 0.1 ml to 1000 kl, preferably from 1 ml.
1000 kl. The residual amount of curcumin after the reaction of the composition containing the microorganism and the curcumin is 99% to 0.001% with respect to the curcumin before the reaction, preferably 95%.
% To 0.001%. The amount of tetrahydrocurcumin produced after the reaction between the microbial cells and the curcumins is 1 mg to 1 g, preferably 10 mg to 1 g per 1 g of curcumin before the reaction.

According to the present invention, tetrahydrocurcumin is produced in a composition containing curcumin substantially free of tetrahydrocurcumin, resulting in a composition containing tetrahydrocurcumin. The present invention may be used for the purpose of increasing the content of tetrahydrocurcumins in the composition containing them and reducing the yellow color of curcumins.

The present invention is intended not only for the purpose of producing substantially pure tetrahydrocurcumin from a reaction product of a microorganism and a curcumin or a composition containing curcumin, but also for a microorganism containing tetrahydrocurcumin or a treated product thereof. May be used as a composition containing tetrahydrocurcumins for use as foods, alcoholic beverages, food additives, medicines, animal feeds, marine feeds, animal drugs, antioxidants, cosmetics, and raw materials thereof. Examples of the treated product include those obtained by treating microbial cells by ultrasonic crushing, heating, organic solvent treatment, pressurization, surfactant treatment, freeze-drying, or the like.

Turmeric is a dry powder of the 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 widely used in food production as safe edible natural pigments. Therefore, a composition containing curcumins purified from turmeric crushed plants, extracts, fractions or turmeric is used as a composition containing curcumins used in the present invention, and is fermented as another production raw material. By using microorganisms and medium components commonly used for production, food, alcoholic beverages, food additives, medicines, animal feed, marine feed, animal drugs, antioxidants, cosmetics and suitable for use as their raw materials,
An extremely safe composition containing tetrahydrocurcumins can be obtained.

Even when a purified or concentrated product of tetrahydrocurcumin is produced from the composition containing tetrahydrocurcumin, the contaminants derived from the reaction solution are only edible components and reaction products of edible microorganisms.
Food, liquor, food additives, medicine, animal feed, fishery feed,
It is possible to obtain a highly safe purified or concentrated tetrahydrocurcumin suitable for use as an animal drug, an antioxidant, a cosmetic, and a raw material thereof.

According to the method of the present invention, various compositions containing tetrahydrocurcumins, such as foods, alcoholic beverages, food additives, medicines, animal feeds, marine feeds, animal drugs, antioxidants, cosmetics, and their raw materials are used. 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 crushed turmeric and tetrahydrocurcumins that act yeast, containing tetrahydrocurcumin that acts turmeric and yeast Production of curry powder to make, production of yeast food containing tetrahydrocurcumin to act turmeric and yeast, yeast containing tetrahydrocurcumin obtained by acting curcumin and yeast to edible oil such as butter, salad oil or sesame oil Production of edible oil containing tetrahydrocurcumin to be immersed, production of soft drink containing tetrahydrocurcumin obtained by allowing turmeric and yeast to act, and the like can be mentioned.

Turmeric tea containing tetrahydrocurcumins can be produced by reacting microbial cells, cultures, or processed products of microorganisms having an activity of converting curcumins into tetrahydrocurcumins with crushed turmeric. Curry powder containing tetrahydrocurcumins is obtained by allowing microbial cells having the activity of converting curcumins to tetrahydrocurcumins, culture solution or a processed product thereof to act on a composition containing curcumins or turmeric. By using a composition containing tetrahydrocurcumins as a raw material of curry powder, curry powder having an improved antioxidant function and pharmacological effect derived from curcumins in the 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 the curry powder is due to the curcumin contained in the turmeric of the curry powder raw material.Therefore, the turmeric in the curry powder is used for the purpose of producing curry powder with enhanced effects other than coloring and flavoring among curcumins. It is not practical to increase the amount of curcumin or to increase the amount of curcumin, because it leads to unnecessarily coloring and flavoring. However, when curcumin or turmeric with an increased content of tetrahydrocurcumin in the present invention is used as a raw material of curry powder, the curry is curried because the yellow color is reduced according to the content of colorless and odorless tetrahydrocurcumin. The functions possessed by the tetrahydrocurcumins can be imparted without exceeding the range of the degree of coloring and the fragrance permitted as powder. However, the composition obtained according to the present invention may be added in addition to the usual curry powder composition as a substitute for turmeric, as long as it is within the edible range.

The yeast food containing tetrahydrocurcumin is prepared by reacting an edible yeast having an activity of converting curcumin into tetrahydrocurcumin on a composition containing curcumin or turmeric to produce tetrahydrocurcumin, By recovering the whole reaction product by freeze-drying or hot-air drying, it is possible to obtain a new yeast food product having a function of a tetrahydrocurcumin added to a yeast food product conventionally produced for nutritional purposes.

An edible yeast having an activity of converting curcumins into tetrahydrocurcumins is allowed to act on a composition containing curcumins or turmeric to generate tetrahydrocurcumins, and then centrifuged or the like to produce tetrahydrocurcumin. After collecting the yeasts containing the yeasts, the yeast foods containing the tetrahydrocurcumins can also be produced by drying the yeasts. Furthermore, the extract obtained by subjecting the yeast containing tetrahydrocurcumin produced in the present invention to hot water extraction or alcohol extraction and then drying or concentrating is compared with yeast extract conventionally produced for nutritional purposes. It can be used as a new edible yeast extract with a class of functions.

The edible oil containing tetrahydrocurcumins is obtained by allowing cells, cultures or processed products of microorganisms having an activity of converting curcumins to tetrahydrocurcumins to a composition containing curcumins or turmeric. After the tetrahydrocurcumin is produced, the edible oil is mixed with the edible oil, and then the edible oil is recovered by a method such as centrifugation, whereby an edible oil containing the tetrahydrocurcumin can be produced. 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 cause air oxidation, and has various physiological functions of tetrahydrocurcumins. It can be used as edible oil.

The alcoholic beverages containing tetrahydrocurcumins are obtained by reacting microbial cells, culture solutions or processed products thereof having the activity of converting curcumins into tetrahydrocurcumins on compositions containing curcumins or turmeric. After producing tetrahydrocurcumins, the alcohol is mixed with beverage alcohol, and then the alcohol is recovered by a method such as centrifugation.If necessary, sweeteners, amino acids, saccharides, coloring agents, flavorings are added, and the alcohol is further added. When it is necessary to increase the concentration, an alcoholic beverage containing tetrahydrocurcumins can be produced by adding ethanol.

Grape or rice is fermented using wine yeast or sake yeast having an activity of converting curcumin into tetrahydrocurcumin, and curcumin or turmeric is added during or at the end of fermentation to add tetrahydrocurcumin. Is produced and then squeezed to produce wine or sake containing tetrahydrocurcumins. Anti-atherosclerotic activity and the like derived from the antioxidant activity of red pigments and tannins contained in red wine has attracted attention, but according to the present invention, white wine with enhanced antioxidant activity without affecting color and taste, and Sake can be manufactured.

Furthermore, milk or the like is fermented using lactic acid bacteria having an activity of converting curcumin into tetrahydrocurcumin, and curcumin or turmeric is added during or at the end of fermentation to produce tetrahydrocurcumin. Fermented milk products containing tetrahydrocurcumins can be produced. As the microorganism used in the present invention, any microorganism can be used as long as it has the activity of converting curcumins into tetrahydrocurcumins. Examples of the microorganism include bacteria, actinomycetes, yeasts, filamentous fungi, mushrooms, and algae.

Examples of the bacteria include Acetobacter and Achromobacte.
r ), the genus Arthrobacter , the genus Bacillus , the 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 ), genus Halobacterium ( Halobacterium ), genus Klebsiella ( Klebsiella ), genus Leuconostoc ( Le
uconostoc ), Micrococcus , Pediococcus , Propionibacterium , Protaminobacter , Providencia
a), the genus Pseudomonas (Psudomonas), Serratia (Serratia), Streptomyces genus (Streptobac
terium), Streptococcus (Streptococcus),
Xanthomonas genus ( Xanthomonas ), Zymomonas genus ( Zy
momonas ), Lactobacillus , Lactobacillus , Pediococcus , Staphylococcus and the like.

Examples of actinomycetes include the genus Actinoplanes and the genus Micromonospora.
pora), Streptomyces (Streptomyces), and the like.

As the yeast, for example, the genus Debaryomyces and the genus Saccharomy
ces), Pichia (Pichia k), Kluyveromyces (Kluyveromyces), Torulaspora genus (Torulaspora
), Candida , Ashby
a ), the genus Brettanomyces , the genus Cryptococcus , the genus Eremothecium , the genus Issatchenki
a ), genus Klockera , lipomyces ( Lip
omyces ), Metschnikowia
), Rodotorua genera (Rhodotorula), Schizosaccharomyces Roma Ise genus (Shizosaccharomyces), Gigot Saccharomyces genus (Zygosaccharomyces), and the like.

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

[0039] Examples of the mushrooms, for example Agaricus genus (Ag
aricus), Aguroshibe genus (Agrocybe), Arumiraria genus (Armillaria), Aurikuraria genus (Auricularia),
Flammulina , Ganoderma ( Ganoderm )
a ), genus Grifola , genus Hypsizigus , genus Irpex , genus Lentinula , genus Lepista , genus Lyophyllum , genus Mycoleptodonoides , namaetro Genus ( Naematol
oma ), Panellus , Poliota ( Pholio )
ta ), genus Pleurotus , genus Pycnoporus , genus Tremella , genus Tricholoma , genus Volvariell
a ) and the like.

As the algae, for example, the genus Analipus ( Anar
ipus ), Condrus ( Chondrus ), Aisenia ( 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 , genus Spirulina , genus Undaria and the like.

Specifically, yeasts such as the genus Devaryomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, lactic acid bacteria such as Lactobacillus and Pecoccus, and grape such as Staphylococcus. Microorganisms such as streptococci can be mentioned. More specifically preferred strains are as follows.

[0042] Debaryomies Hansenii ( Debaryom)
yces hansenii ) ATCC-20261, Debaryomyces hansenii IFO-0094,

Saccharomyces cerevisiae ( Saccha
romyces cerevisiae ) Saccharomyces cerevisiae (manufactured by dry yeast Lallmand)
IAM-4500, Saccharomyces cerevisiae ( Saccharo
myces cerevisiae ) IAM-4519, Saccharomyces cerevisiae ATCC-7754,
Saccharomyces cere
visiae ) FERM-P-6189, Saccharomyces cerevisiae ( Saccharomyces cerevisiae ) IFO-2044, Saccharomyces cerevisiae ( Saccharomyces cerevisiae )
ATCC-20018, Saccharomyces cerevisiae ( Sacc
haromyces cerevisiae FERM-P-6213, Saccharomyces cerevisiae FERM-
P-6214, Saccharomyces cerevisiae ( Saccharomy
ces cerevisiae FERM-P-7614, Saccharomyces cerevisiae FERM-P-761
Five,

Pichia subpericulosa
pelliculosa ) ATCC- 16766 , Pichia Anomara ( Pich
ia anomala) ATCC-2149, Pichia Men bra seedlings Faye tumefaciens (Pichia membranaefaciens) IAM-4986, Pichia Men bra seedlings Faye tumefaciens (Pichia membranaefac
iens) ATCC-36908, Pichia kluyveri (Pichia klu
yveri ) ATCC-9768,

Kluyveromyces Mark Cyanus ( Kl
uyveromyces marxianus ) IFO-0433, Kluyveromyces marxianus ( Kluyveromyces marxianus ) IFO-
1090, Kluyveromyces polysporus
ces polysporus ) ATCC-22028,

[0046] Torulasp
ora delbrueckii ) IAM-4816,

[0047] Candida utilis
ATCC-9950, Candida famat
a ) ATCC-2560,

Lactobacillus plantarum ( Lactobac)
illus plantarum ) Kyowa High Foods LPT, Staphylococcus carnosus
TEXEL manufactured by M72, Pedjiokokkasu-Ashidjirakuchishi (Pe
diococcus acidilactici ) TEXEL P2M120, Staphylococcus xylosus 2 (T)
EXEL P2M120.

Any of these strains can be used alone or in combination. In addition, even when these strains are mutated by artificial mutation methods, for example, ultraviolet irradiation, X-ray irradiation, treatment with a mutagenic agent, genetic manipulation or the like, or naturally mutated, the curcumins are converted to tetrahydrocurcumins. Any competent strain can be used in the present invention.

For culturing these microorganisms, a natural medium containing a carbon source, a nitrogen source and other nutrients, a semi-synthetic medium, and a synthetic medium can be used as long as the medium can grow on a medium used for culturing ordinary yeasts and bacteria. 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, molasses and the like, and these can be used alone or in combination. Furthermore, depending on the assimilation ability of the bacteria, hydrocarbons, alcohols,
Organic acids and the like may be used.

Examples of the nitrogen source include ammonium chloride, ammonium nitrate, ammonium sulfate, sodium nitrate, urea, peptone, meat extract, yeast extract, dried yeast, corn steep liquor, soybean flour, casamino acid and the like. Can be used. Examples of the inorganic salts include sodium chloride, potassium chloride, magnesium sulfate, calcium carbonate, potassium dihydrogen phosphate, ferrous sulfate, calcium chloride, manganese sulfate, zinc sulfate, copper sulfate and the like. These may be used alone or in combination. be able to.

Examples of the trace components include vitamins such as biotin, thiamine and nicotinic acid, and amino acids such as β-alanine and glutamic acid. These can be used alone or in combination. As a culture method,
The liquid culture method, particularly the deep stirring culture method, is suitable. The cultivation is carried out at a temperature of 10 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 takes 1-7 days. Ammonia water or ammonium carbonate solution is used to adjust the pH of the medium.

The treated product of the culture solution used in the present invention includes:
Concentrate, dried, frozen, of a culture of a microorganism having an activity of converting curcumins into tetrahydrocurcumins
Refrigerated, freeze-dried, heated, pressurized, ultrasonically crushed,
A surfactant or an organic solvent-treated product, a lysed enzyme-treated product, and the like can be given. In addition, as the treated cells, dried, frozen, chilled, lyophilized, heated, pressurized, and ultrasonically crushed cells of microorganisms having an activity of converting curcumins into tetrahydrocurcumins. And a surfactant or an organic solvent-treated product, a lysed enzyme-treated product, immobilized cells or an enzyme extracted from the cells.

For purification of the enzyme from the cells, a general protein purification method can be used. For example, homogenizers, glass beads, dissolution of ammonia, crushing of cells by enzymatic method, filtration, collection of enzyme solution by filtration, centrifugation, salting out, organic solvent precipitation, collection of enzymes by antibodies, concentration by dialysis, etc. External filtration, gel filtration, electrophoresis, separation by liquid phase distribution, ion exchanger, adsorbent,
Chromatography using an affinity adsorbent, a batch method, crystallization and the like can be used alone or in combination.

Curcumins can be obtained as a reagent, for example, from Sigma-Aldrich Japan. The composition containing curcumins is not particularly limited, and examples thereof include a composition obtained by treating a plant belonging to the genus Turmeric. Plants belonging to the genus Turmeric include, for example, Kyou-oh (commonly called spring turmeric) and turmeric (commonly referred to as spring turmeric).
Called autumn turmeric. ) And gejutsu. The treatment method is not particularly limited, and examples thereof include methods such as pulverization, drying, and extraction. Examples of the composition containing curcumins include common foods such as curry powder and turmeric tea.

In addition, curcumin-free foods, alcoholic beverages, feeds, and cosmetics may be used in addition to curcumin or curcumin-containing foods, alcoholic beverages, feeds, and cosmetics. Curcumin-free foods, alcoholic beverages, feed,
Examples of the cosmetic include alcohol for drinking, edible oil, fruit juice and the like. Curcumins and curcumin-containing compositions can be easily obtained as commercial products as foods.

Hereinafter, the present invention will be described by way of examples, which are illustrative and do not limit the present invention. The following analyzers were used for the test results shown below. Mass spectrometry (instrument: JMS-HX110 / 110A (JEOL), FAB gas: X
e ^o , pressurized voltage: 10kV, matrix: m-nitrobenzyl alcho
l), nuclear magnetic resonance method (Equipment: JNM-A400 (JEOL, 400MHz for
¹ H-NMR, 100 MHz for ¹³ C-NMR), solvent: CDCl ₃ )

[0058]

EXAMPLE 1 Conversion of Curcumin to Tetrahydrocurcumin by Yeast Debaryomyces hansenii ATCC-20261 Strain Debaryomyces hansenii TCC-20261 Strain TCC-20261 Medium 20 g / L in YM Medium (DIFCO) in a 20 ml test tube
The resulting mixture was inoculated into a ml culture and shake-cultured (130 rpm) at 30 ° C. for 24 hours to obtain a seed medium. The seed medium was inoculated at a ratio of 5% into 300 ml of YM medium in a 2 liter Erlenmeyer flask, and cultured with shaking at 30 ° C. for 24 hours. 300 ml of the obtained culture was centrifuged at 10,000 rpm for 20 minutes, and the supernatant was removed to obtain cells. The cells were suspended in water in a 300 ml Erlenmeyer flask to make a total volume of 30 ml, and 1.5 mg of a 4 mg / ml curcumin solution (solvent: ethanol) was added to the suspension to prepare a reaction solution. The reaction solution was added to 30
The mixture was shaken at 3 ° C. for 3 hours. After 3 hours, the reaction solution was extracted with 100 ml of ethyl acetate to obtain an ethyl acetate extract.

When the ethyl acetate extract was analyzed by HPLC under the following conditions, peaks at a retention time of 6.0 minutes and 11.4 minutes were observed. The peaks coincided with the retention times of 6.1 minutes and 11.4 minutes of the tetrahydrocurcumin preparation (manufactured by Niken Kasei) under the same conditions.

HPLC analysis conditions Column: ODS 120A S-5 (AM312, manufactured by YMC) Solvent: acetonitrile-water (50:50, V / V) (including 0.05%
(Trifluoroacetic acid) Flow rate: 1 ml / min Detection: 280, 430 nm Retention time 6.0 min and 1 min by HPLC analysis of the ethyl acetate extract
The compound eluted at 1.4 minutes was isolated as follows. After concentrating the ethyl acetate extract, it was developed and fractionated with an n-hexane-ethyl acetate (10% stepwise) mixed solution using a silica gel column (Wakogel C-200, manufactured by Wako Pure Chemical Industries, Ltd.). The eluted 50% and 60% ethyl acetate-n-hexane fractions were concentrated, and this was concentrated to n-hexane-ethyl acetate (1: 1).
Fractionation by preparative thin-layer chromatography using 1, V / V) as a developing solvent gave a fraction having an Rf value of 0.4 to 0.5. The fraction was fractionated by preparative thin-layer chromatography using n-hexane-acetone (2: 1, V / V) as a developing solvent to give an Rf value of 0.25.
Yielded a fraction of 0.3. The fraction was further fractionated by preparative thin-layer chromatography using toluene-acetone (3: 1, V / V) as a developing solvent to obtain a fraction having an Rf value of 0.5, and this fraction was concentrated and dried. By solidification, 2.9 mg of compound was obtained.

Instrumental analysis results of isolated compound High resolution FAB mass spectrum (positive mode; matrix: m-NBA): m / z aum 372. 1555 Δ-1.8 mmu (as C ₂₁ H ₂₄ O ₆ ) ¹ H NMR spectrum [400 MHz, CDCl ₃ solution, 30
° C] δppm [Multiplicity, coupling constant (Hz)] 6.82 (d, 8.3), 6.6
8 (s), 6.66 (m), 3.86 (s), 3.50 (s), 2.85 (t, 7.8), 2.55
(t, 7.8) ¹³ C NMR spectrum [100 MHz, CDCl ₃ solution, 30
° C] δppm (multiplicity) 193.2 (s), 146.5 (s), 144.1 (s), 132.
6 (s), 129.4 (s), 120.9 (d), 115.4 (d), 114.0 (d), 111.
0 (d), 99.8 (d), 57.7 (d), 55.9 (q), 45.5 (t), 40.4 (t),
31.4 (t), 29.2 (t)

Instrumental analysis result of tetrahydrocurcumin sample High-resolution FAB mass spectrum (positive mode; matrix: m-NBA): m / z aum 372. 1562 Δ-1.1 mmu (as C ₂₁ H ₂₄ O ₆ ) ¹ HNMR spectrum [400 MHz, CDCl ₃ solution, 30
° C] δppm [Multiplicity, coupling constant (Hz)] 6.82 (d, 8.1), 6.6
7 (s), 6.66 (m), 3.85 (s), 3.50 (s), 2.84 (t, 7.8), 2.54
(t, 7.8) ¹³ C NMR spectrum [100 MHz, CDCl ₃ solution, 30
° C] δppm (multiplicity) 193.2 (s), 146.5 (s), 144.1 (s), 132.
6 (s), 129.4 (s), 120.9 (d), 115.4 (d), 114.0 (d), 111.
0 (d), 99.8 (d), 57.7 (d), 55.9 (q), 45.5 (t), 40.4 (t),
31.3 (t), 29.2 (t)

The results of mass spectrometry and nuclear magnetic resonance spectrum of this compound were consistent with those of the tetrahydrocurcumin standard.
This compound was confirmed to be tetrahydrocurcumin. In the above conversion reaction, the incubation time was 0.5, 1,
Table 1 shows the conversion rates of curcumin to tetrahydrocurcumin at 2 and 4 hours. The concentration was determined by HPLC conversion of the concentration in the reaction solution from the standard product.
The conversion rate is represented by the product tetrahydrocurcumin concentration / added curcumin concentration × 100 (%).

[0064]

[Table 1]

Example 2 Conversion of Curcumin in Turmeric into Tetrahydrocurcumin by Yeast Debaryomyces hansenii TCC-20261 Strain Debaryomyces hansenii ATCC-20261 was used as a seed for the conversion reaction. The strain was inoculated into 50 ml of a medium consisting of 21 g / l of YM medium (DIFCO) in a 20 ml test tube, and cultured with shaking (130 rpm) at 30 ° C. for 24 hours to obtain a seed medium. YM medium in a 2 liter Erlenmeyer flask
The cells were inoculated at a ratio of 5% into 300 ml and cultured with shaking at 30 ° C. for 24 hours. 300 ml of the obtained culture was centrifuged at 10,000 rpm for 20 minutes, and the supernatant was removed to obtain cells. The cells were suspended in water in a 300 ml Erlenmeyer flask to a total volume of 30 ml, and a 332 mg / ml turmeric solution (solvent: ethanol) was added to the suspension.
1.5 ml was added to obtain a reaction solution. The reaction solution is heated at 30 ° C for 3 hours.
After shaking for 3 hours, the reaction solution was
Extraction with 1 gave an ethyl acetate extract.

When the ethyl acetate extract was analyzed by HPLC under the following conditions, peaks at a retention time of 6.0 minutes and 11.4 minutes were observed. The peaks coincided with the retention times of 6.1 minutes and 11.4 minutes of the tetrahydrocurcumin preparation (manufactured by Niken Kasei) under the same conditions.

HPLC analysis conditions Column: ODS 120A S-5 (AM312, manufactured by YMC) Solvent: acetonitrile-water (50:50, V / V) (including 0.05%
(Trifluoroacetic acid) Flow rate: 1 ml / min Detection: 280 nm, 430 nm

Example 3 Ability of Converting Curcumin in Turmeric from Various Microorganisms to Tetrahydrocurcumin The strains shown in Tables 2 and 3 were used. Microorganisms belonging to the genus Devaryomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, and Candida were isolated from YM medium (manufactured by DIFCO) in test tubes having a capacity of 20 ml.
The cells were inoculated into 5 ml of a medium containing 1 g / l, and cultured with shaking (130 rpm) at 30 ° C. for 24 hours. LPT, a commercial lactic acid bacteria starter
(Manufactured by Kyowa High Foods), M72 (manufactured by TEXEL) and P2
M120 (manufactured by TEXEL) was used for LS medium (1.25% yeast extract, 1.25% polypeptone, 1.1% glucose, 1.5% maltose, 0.025% K ₂ HPO ₄ , KH ₂ PO ₄
0.025%, ₂ O 1.0% Sodium acetate _{· 3H, MgSO 4 · 7H 2} O 0.0
_{1%, MnSO 4 · 4H 2} O 0.0005%, FeSO 4 0.0005%, Tween80
0.03%, L-cysteine / HCl 0.03%, adenine 0.002%,
5 ml of uracil (0.002%) was inoculated and cultured at 37 ° C. for 24 hours.

5 ml of the obtained culture was centrifuged at 10,000 rpm for 20 minutes, and the supernatant was removed to obtain cells. The cells were suspended in a 1.5 ml polypropylene tube, and the total volume was
The suspension is added with 25 μl of a 332 mg / ml or 33.2 mg / ml turmeric solution (solvent: ethanol) (curcumin concentration is 4 mg / ml and 0.4 mg / ml, respectively). And reacted. After 3 hours, the whole reaction solution containing the cells was extracted with 1 ml of ethyl acetate to obtain an ethyl acetate extract. The ethyl acetate extract was concentrated to dryness under reduced pressure, dissolved in 100 μl of dimethyl sulfoxide,
l was analyzed by HPLC. The tetrahydrocurcumin content was calculated by comparing the area ratio of the HPLC peak with the tetrahydrocurcumin standard.

The concentration of added curcumin is the initial concentration of curcumin in the reaction solution (μg / ml), and the concentration of formed tetrahydrocurcumin is the concentration of tetrahydrocurcumin after incubation (μg / ml)
Was determined by HPLC conversion from the standard product. The conversion rate is the concentration of the produced tetrahydrocurcumin / the concentration of added curcumin × 100
(%). As a result, as shown in Tables 2 and 3, it was shown that tetrahydrocurcumin was produced at various concentrations.

[0071]

[Table 2]

[0072]

[Table 3]

Example 4 Mass Production of a Composition Containing Tetrahydrocurcumin Using Turmeric as a Raw Material (Using the Yeast Devaryomyces hansenii ATCC-20261 Strain)
The 0261 strain was used. The strain was placed in a 2 liter Erlenmeyer flask.
YM medium (DIFCO) was inoculated into 300 ml of a medium containing 21 g / l, and cultured with shaking (130 rpm) at 30 ° C. for 48 hours. YM in a 30-liter jar fermenter was prepared.
The medium was inoculated at a rate of 5% (volume) into 18 liters of a medium consisting of 21 g / l of the medium, and cultured under aeration and agitation (rotation speed: 250 rpm, aeration rate: 18 liters / minute) at 28 ° C.

After culturing for 24 hours, cells were obtained using a continuous centrifugal separator, and the cells were suspended in water to a total volume of 1.8 liters. A 332 mg / ml turmeric solution (solvent:
Ethanol (90 ml) was added and stirred at 28 ° C. for 3 hours to react. After 3 hours, the reaction solution is freeze-dried,
Yeast powder containing 4 mg of tetrahydrocurcumin per gram was obtained.

Example 5 Mass Production of a Composition Containing Tetrahydrocurcumin Using Curcumin as a Raw Material (Staphylococcus carnosus TEXE as a starter for lactic acid bacteria)
L-type M72) Lactobacillus starter, Staphylococcus carnosus TEXEL M72, was used as the inoculum. The strain was inoculated into 300 ml of a medium containing 21 g / l of YM medium (DIFCO) in a 2 liter Erlenmeyer flask, and cultured at 37 ° C. for 48 hours. The seed culture solution thus obtained was inoculated into 2 liters of LS medium in a 3 liter Erlenmeyer flask at a ratio of 5% (volume), and cultured at 37 ° C. for 48 hours (10 tubes). After culturing for 24 hours, cells were obtained using a continuous centrifuge, and the cells were suspended in water to a total volume of 2 liters. 100 ml of a 33.2 mg / ml turmeric solution (solvent: ethanol) was added to this suspension, and the mixture was stirred at 28 ° C. for 3 hours to react. After 3 hours, the reaction solution was freeze-dried to obtain lactic acid bacteria powder containing 1 mg / g of tetrahydrocurcumin.

Example 6 Example of Production of Tetrahydrocurcumin Using Crude Enzyme Extracted from Bacteria (Using Yeast Debaryomyces hansenii ATCC-20261) 300 ml of Debaryomyces hansenii ATCC-20261 was used.
Inoculate 50 ml of a medium consisting of 21 g / l of YM medium (DIFCO) in a Erlenmeyer flask with a capacity and shake culture at 30 ° C for 24 hours (13
0 rpm) to obtain a culture solution. The culture was incubated at 10,000 rpm for 20 minutes.
After centrifugation for minutes, the cells were obtained by removing the supernatant. Water the cells
The suspension was suspended in 2 ml, centrifuged at 10,000 rpm for 20 minutes, and the supernatant was removed to obtain 0.8 ml of cells. 0.8 ml of the cells were added to an extraction buffer [20 mM Tris-Hcl (pH 7.5), 1 mM EDTA, 5 mM MgC
_{l 2, 50 mM KCl, 5} % glycerol, 1mM phenylmethanes
ulfonyl fluoride], and added thereto 3.2 ml of glass beads. In a cold room, the suspension was shaken vigorously for 30 seconds using a vortex mixer and immediately cooled on ice for 1 minute. This shaking operation and cooling operation
This was repeated twice to disrupt the cells. The crushed suspension of the cells was centrifuged at 15,000 rpm for 30 minutes to remove the precipitate, and 0.8 ml of a supernatant cell extract was obtained.

A reaction solution was prepared by adding 5 μl of a 4 mg / ml curcumin solution (solvent: ethanol) to 100 μl of the bacterial cell extract. The reaction solution was kept at 37 ° C. for 2 hours, and after 2 hours, the reaction solution was extracted with 1 ml of ethyl acetate to obtain an ethyl acetate extract. The ethyl acetate extract was subjected to HPLC under the following conditions.
Analysis revealed peaks with retention times of 6.0 minutes and 11.4 minutes. The peaks coincided with the retention times of 6.1 minutes and 11.4 minutes of the tetrahydrocurcumin preparation (manufactured by Niken Kasei) under the same conditions.

HPLC analysis conditions Column: ODS 120A S-5 (AM312, manufactured by YMC) Solvent: acetonitrile-water (50:50, V / V) (including 0.05%
(Trifluoroacetic acid) Flow rate: 1 ml / min Detection: 280 nm, 430 nm

Example 7 Production of Tetrahydrocurcumin-Containing Edible Oil (1) Yeast Debaryomyces hansenii ATCC-20261 strain was inoculated into 300 ml of 21 g / l YM medium (DIFCO) in a 2 liter Erlenmeyer flask. Shaking culture (130 rpm) was performed at 30 ° C. for 48 hours. The obtained seed culture solution was inoculated at a rate of 5% (volume) into 18 liters of a medium comprising 21 g / l of YM medium in a jar fermenter having a capacity of 30 liters at a rate of 5% (volume), and aerated and stirred at 28 ° C. (Aeration rate of 18 liters / minute). After culturing for 24 hours, cells were obtained using a continuous centrifuge, and the cells were suspended in water to a total volume of 1.8 liters. 332m to this suspension
90 ml of a g / ml turmeric solution (solvent: ethanol) was added, and the mixture was stirred and reacted at 28 ° C. for 3 hours. After 3 hours, 1 liter of sesame oil was added to the reaction solution, and the mixture was further stirred for 6 hours to extract tetrahydrocurcumin into sesame oil. This was allowed to stand for 2 hours to separate an aqueous layer and an oil layer. After collecting the oil layer, the oil layer was filtered to obtain 1 liter of sesame oil containing tetrahydrocurcumin. Almost no coloring due to unreacted curcumin was observed.

The sesame oil fraction was developed and fractionated with an n-hexane-ethyl acetate (10% stepwise) mixed solution using a silica gel column (Wakogel C-200, manufactured by Wako Pure Chemical Industries, Ltd.). The eluted 50% and 60% ethyl acetate-n-hexane fractions were concentrated and analyzed by HPLC under the following conditions. As a result, peaks with a retention time of 6.0 minutes and 11.4 minutes were observed. The peaks coincided with the retention times of 6.1 minutes and 11.4 minutes of the tetrahydrocurcumin preparation (manufactured by Niken Kasei) under the same conditions.

HPLC analysis conditions Column: ODS 120A S-5 (AM312, manufactured by YMC) Solvent: acetonitrile-water (50:50, V / V) (including 0.05%
(Trifluoroacetic acid) Flow rate: 1 ml / min Detection: 280 nm, 430 nm

Example 8 Production of Tetrahydrocurcumin-Containing Edible Oil (2) 10 g of the tetrahydrocurcumin-containing dry yeast obtained by the method of Example 4 was placed in a 3 liter Erlenmeyer flask, and 1 liter of sesame oil was added. After stirring for 6 hours, tetrahydrocurcumin was extracted into sesame oil. This was filtered to obtain 1 liter of sesame oil containing tetrahydrocurcumin. Almost no coloring due to unreacted curcumin was observed.

The sesame oil fraction was developed and fractionated with an n-hexane-ethyl acetate (10% stepwise) mixed solution using a silica gel column (Wakogel C-200, manufactured by Wako Pure Chemical Industries, Ltd.). The eluted 50% and 60% ethyl acetate-n-hexane fractions were concentrated and analyzed by HPLC under the following conditions. As a result, peaks with a retention time of 6.0 minutes and 11.4 minutes were observed. The peaks coincided with the retention times of 6.1 minutes and 11.4 minutes of the tetrahydrocurcumin preparation (manufactured by Niken Kasei) under the same conditions.

HPLC analysis conditions Column: ODS 120A S-5 (AM312, manufactured by YMC) Solvent: acetonitrile-water (50:50, V / V) (including 0.05%
(Trifluoroacetic acid) Flow rate: 1 ml / min Detection: 280 nm, 430 nm

Example 9 Production of Sake Containing Tetrahydrocurcumin (1) Yeast Debaryomyces hansenii ATCC-20261 strain was inoculated into 300 ml of a medium containing 21 g / l of YM medium (DIFCO) in a 2 liter Erlenmeyer flask. Shaking culture (130 rpm) was performed at 48 ° C for 48 hours. The obtained seed culture solution was inoculated at a rate of 5% (volume) into 18 liters of a medium comprising 21 g / l of YM medium in a jar fermenter having a capacity of 30 liters at a rate of 5% (volume), and aerated and stirred at 28 ° C. (Aeration rate of 18 liters / minute). After culturing for 24 hours, cells were obtained using a continuous centrifuge, and the cells were suspended in water to a total volume of 1.8 liters. 332m to this suspension
90 ml of a g / ml turmeric solution (solvent: ethanol) was added, and the mixture was stirred and reacted at 28 ° C. for 3 hours. Three hours later, 1 liter of 95% ethanol was added to the reaction solution, and then a further 6 hours.
Stirred for hours. This was filtered to obtain 2.8 liters of a liquor containing tetrahydrocurcumin. Almost no coloring due to unreacted curcumin was observed.

When the ethyl acetate extract was analyzed by HPLC under the following conditions, peaks with a retention time of 6.0 minutes and 11.4 minutes were observed. The peaks coincided with the retention times of 6.1 minutes and 11.4 minutes of the tetrahydrocurcumin preparation (manufactured by Niken Kasei) under the same conditions.

HPLC analysis conditions Column: ODS 120A S-5 (AM312, manufactured by YMC) Solvent: acetonitrile-water (50:50, V / V) (containing 0.05%
(Trifluoroacetic acid) Flow rate: 1 ml / min Detection: 280 nm, 430 nm

Example 10 Production of Tetrahydrocurcumin-Containing Sake (2) 10 g of the tetrahydrocurcumin-containing dry yeast obtained by the method of Example 4 was placed in a 3-liter Erlenmeyer flask, and 1 liter of shochu having an alcohol content of 25 degrees was added. After stirring for 6 hours,
Tetrahydrocurcumin was extracted. This was filtered to obtain 990 ml of shochu containing tetrahydrocurcumin. Almost no coloring due to unreacted curcumin was observed.

The shochu containing tetrahydrocurcumin was analyzed by HPLC under the following conditions, and peaks having a retention time of 6.0 minutes and 11.4 minutes were observed. Under the same conditions, the retention time of tetrahydrocurcumin standard (manufactured by Niken Kasei) was 6.1 minutes and
This coincided with the peak at 11.4 minutes.

HPLC analysis conditions Column: ODS 120A S-5 (AM312, manufactured by YMC) Solvent: acetonitrile-water (50:50, V / V) (including 0.05%)
(Trifluoroacetic acid) Flow rate: 1 ml / min Detection: 280 nm, 430 nm

Example 11 Production of Tetrahydrocurcumin-Containing Sake (3) 10 g of tetrahydrocurcumin-containing dry yeast obtained by the method of Example 4 was placed in a 3 liter Erlenmeyer flask, and 1 liter of white wine was added. For 6 hours to extract tetrahydrocurcumin. This was filtered to obtain 990 ml of white wine containing tetrahydrocurcumin. Almost no coloring due to unreacted curcumin was observed.

After the tetrahydrocurcumin-containing white wine was concentrated, it was analyzed by HPLC under the following conditions.
Min and 11.4 min peaks were observed. The peaks coincided with the retention times of 6.1 minutes and 11.4 minutes of the tetrahydrocurcumin preparation (manufactured by Niken Kasei) under the same conditions.

HPLC analysis conditions Column: ODS 120A S-5 (AM312, manufactured by YMC) Solvent: acetonitrile-water (50:50, V / V) (containing 0.05%
(Trifluoroacetic acid) Flow rate: 1 ml / min Detection: 280 nm, 430 nm

[0094]

According to the present invention, curcumin or a composition containing curcumin is reacted with a microorganism which catalyzes a hydrogenation reaction using curcumin as a substrate to produce a composition containing tetrahydrocurcumin. be able to.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI (C12P 7/26 C12R 1:84) (C12P 7/26 C12R 1:84) (C12P 7/26 C12R 1: 865) (C12P 7/26 C12R 1:01 1:44)

Claims (14)

[Claims] 1. A compound of the formula (I) [In the formula (I), R ¹ , R ² , R ³ and R ⁴ are the same or different and each represents hydrogen, hydroxy or lower alkoxy] (hereinafter, simply referred to as curcumins). II) [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]
A microbial cell, a culture solution or a treated product thereof, which has an activity of converting to a tetrahydrocurcumin), is allowed to act on curcumin or a composition containing curcumin to produce tetrahydrocurcumin, and the resulting tetrahydrocurcumin is produced. A method for producing tetrahydrocurcumins, which comprises collecting the same. 2. A method of producing a tetrahydrocurcumin by reacting a microbial cell, a culture solution or a treated product thereof with a composition containing the curcumin, which has an activity of converting the curcumin into a tetrahydrocurcumin. A method for producing a composition containing tetrahydrocurcumins. 3. adding curcumin or a composition containing curcumin to a composition not containing curcumin;
Manufacture of a composition containing tetrahydrocurcumin, wherein the composition is produced by reacting a microorganism having an activity of converting curcumin into tetrahydrocurcumin, a culture solution or a processed product thereof to produce tetrahydrocurcumin. Method. 4. The method according to claim 1, wherein the composition containing curcumins is a composition obtained by treating a plant belonging to the genus Turmeric. 5. The method according to claim 3, wherein the curcumin-free composition is a beverage alcohol, fruit juice or edible oil. 6. The method according to claim 3, wherein the curcumin-free composition is an organic solvent or a surfactant. 7. The microorganism according to claim 1, wherein the microorganism belongs to the genus Devaryomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, Lactobacillus, Staphylococcus, or Pecoccus. Any of the methods described. 8. A method for producing a microorganism containing tetrahydrocurcumins, which comprises culturing a microorganism having an activity of converting curcumins into tetrahydrocurcumins in a medium containing curcumins. 9. A microorganism having an activity of converting curcumins into tetrahydrocurcumins, wherein the microorganism is Debaryomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, Lactobacillus, Staphylococcus, 9. The method according to claim 8, which is a microorganism belonging to the genus Pecoccus. 10. A microorganism containing tetrahydrocurcumins in cells. 11. The microorganism according to claim 10, wherein the microorganism belongs to the genus Devaryomyces, Saccharomyces, Pichia, Kluyveromyces, Torulaspora, Candida, Lactobacillus, Staphylococcus, or Pecoccus. Microorganisms. 12. A method for producing an edible oil containing tetrahydrocurcumins, which comprises immersing a microorganism containing tetrahydrocurcumins or a processed product thereof in edible oil. 13. A method for producing a liquor containing tetrahydrocurcumin, which comprises immersing a microorganism containing tetrahydrocurcumin or a processed product thereof in liquor. 14. The production method according to claim 12 or 13, wherein the microorganism containing tetrahydrocurcumins or a treated product thereof is obtained by the method according to claim 8.