JP3151363B2 - Method for producing trans-2-hexenal-containing composition, and perfume composition containing the composition - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing a trans-2-hexenal-containing composition having a preferable green note, a method for producing trans-2-hexenal, and a fragrance composition containing the composition.

[0002]

2. Description of the Related Art Trans-2-hexenal is a very strong green notebook [= blue scent, Japan Fragrance Association, “Encyclopedia of Fragrances” (June 25, 1989, published by Asakura Shoten Co., Ltd.)
on page xi), which are widely used as fragrances for imparting green notes to apple and grape flavors. Fragrances are roughly classified into two types depending on their raw materials or manufacturing methods: flavors of chemically synthesized products (so-called "synthetic flavors") and flavors other than chemically synthesized products (so-called "natural flavors"). People tend to prefer “natural flavors” to “synthetic flavors”. But transformer-2
-Hexenal is present in trace amounts in nature and is very expensive to extract from natural products, so that in most cases it is produced by chemical synthesis means.

On the other hand, as a natural fragrance having green notes, for example, a composition containing an acyclic alcohol having 6 carbon atoms, which is obtained by distilling mint oil derived from Mentha, is used. I have. This composition is mainly composed of 30 to 55% by weight of trans-2-hexenol, 15 to 50% by weight of cis-3-hexenol and 15 to 30% by weight of n-hexanol, among which cis-3-hexenol is excellent. Has a green note. However, since the odor of trans-2-hexenol is not preferable as green notes, when the composition is used for imparting green notes, trans-2 hexenol is used.
-It is desirable that the content of hexenol be as low as possible.

However, trans-2-hexenol and cis-
Since it is extremely difficult to separate from 3-hexenol, it is also extremely difficult to separate only trans-2-hexenol from the composition. It is also conceivable to selectively oxidize only trans-2-hexenol in the composition to convert it to trans-2-hexenal having a favorable odor, but an effective method for such selective oxidation is known. Not been. Therefore, at present, a fragrance having a more preferable green note has not been obtained from the acyclic alcohol-containing composition having 6 carbon atoms derived from mint oil as described above.

[0005] By the way, Candida Boidini (Ca
ndida boidinii) Strain AOU-1 is to oxidize various lower alcohols such as methanol, ethanol, n-propanol and allyl alcohol to convert them to the corresponding aldehydes in methanol or a medium containing methanol and glycerin. (Yoshiki Tani et al., Ag
ric. Biol. Chem., 49 (9) pp. 2699-2706 (1985) and Yasu
yoshi Sakai et al., Agric. Biol. Chem., 51 (9) pp.2617-26
20 (1987)]. According to the fermentation method using Candida boidini AOU-1 strain, alcohols are oxidized without a coenzyme by a reaction of an enzyme produced by the cells, and aldehydes can be easily obtained. Moreover, the reaction stops at the stage of oxidation to the aldehyde, and no further oxidation reaction proceeds.
For the purpose of obtaining aldehyde, there is an advantage that a step such as stopping the reaction in the middle is not required.

However, according to reports to date,
In the method using the Candida boidini AOU-1 strain, alcohols other than the lower alcohols described above, for example, alcohols having 5 to 8 carbon atoms, are not oxidized at all, or are very small even if oxidized. Also, the transformer
There is no report that an alcohol having a specific configuration such as -2-hexenol is selectively oxidized.

[0007]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is, for example, to selectively oxidize trans-2-hexenol contained in a composition containing trans-2-hexenol to trans-2-hexenal. And a method for producing a composition having more preferable green notes, and a method for producing trans-2-hexenal. Another object of the present invention is to provide a fragrance composition having more preferable green notes.

[0008]

In order to solve the above problems, the present inventors have been known to produce Candida boidini bacterium, which is conventionally known to produce a corresponding aldehyde by oxidizing a lower alcohol such as methanol. Were examined. Candida boidini AOU-1 strain described in Japanese Patent Application Laid-Open No. 61-67480, which is a mutant having parent strain Strain Candida boidini SA051, can produce formaldehyde by culturing in methanol. It is known (Yasuyoshi Sakai et al., Agric. Bio
l. Chem., 51 (8) pp. 2177-2184 (1987)], the present inventors have found that the cells of the Candida boidini SA051 strain or the processed product thereof have a double bond at the carbon at position 2. It has been found that a trans-type acyclic alcohol (trans-2-alkenol) having about 4 to 10 carbon atoms having the following formula is oxidized to be converted to a corresponding aldehyde (trans-2-alkenal). Moreover, when a mixture of alcohols having the same number of carbon atoms is brought into contact with the cells of Candida boidini SA051 strain or a treated product thereof, the oxidation rate of trans-2-alkenols is significantly higher than that of other alcohols. It has been found that trans-2-alkenals can be obtained before the alcohol is oxidized. That is, if such a property of the cells of the Candida boidini SA051 strain or a processed product thereof is used, a plurality of trans-2-hexenol, cis-3-hexenol, n-hexanol and the like obtained from mint oil and the like can be used. It has been found that trans-2-hexenol in a composition containing an acyclic alcohol having 6 carbon atoms can be selectively oxidized to trans-2-hexenal to obtain a composition having a preferable green note, The present invention has been completed.

[0009] The present invention relates to a composition containing trans-2-hexenol containing Candida boidin.
ii) by contacting the cells or the processed product thereof,
Provided is a method for producing a trans-2-hexenal-containing composition, which comprises converting 2-hexenol to trans-2-hexenal. Further, the present invention provides a method for producing trans-2-hexenal, comprising isolating trans-2-hexenal from the trans-2-hexenal-containing composition obtained by the method for producing the composition. I do.

Further, the present invention provides a fragrance composition comprising a trans-2-hexenal-containing composition obtained by the method for producing the composition. The trans-2-hexenol-containing composition which is a starting material of the method for producing the composition of the present invention may be any composition as long as it contains trans-2-hexenol. It is suitable when the composition contains trans-2-hexenol and other alcohols, and particularly, trans-2-hexenol and other acyclic alcohols having 6 carbon atoms (for example, cis-3-hexol). Hexenol, n-hexanol, etc.). Representative examples of compositions containing trans-2-hexenol and other acyclic alcohols having 6 carbon atoms include a composition obtained by distilling mint oil derived from Mentha. Can be mentioned. In addition, for example, grape genus (Vi
tis) fruit essential oil; orange oil, mandarin oil, bergamot oil and their water-soluble fractions obtained by pressing the skin of Rutaceae; water-soluble fraction of apple juice; geranium (Pelargonium inquinan)
s) Geranium oil obtained by steam distillation of leaves and its water-soluble fraction; parsley (Petroselinum sativu)
m) Parsley leaf oil obtained by steam distillation of leaves; Absolute obtained by solvent extraction of flowers of leguminaceae (Genetica, also referred to as bloom; Cytisus scoparius or Genistetinctoria); Narcissus (Na)
rcissusTazetta) can also include a composition derived from a natural product such as an absolute obtained by extracting a flower with a solvent.

The above-mentioned composition obtained by distilling mint oil is obtained by steam distillation of whole plants of the genus Henta (Mentha) such as peppermint and spearmint, and Japanese varieties. About 0.08 in mint oil
~ 0.3% by weight of alcohol content,
Preferably, a fraction obtained by distilling mint oil (a commercially available mint oil can be used as it is) [distillation temperature: 70 to 80 ° C. (13 to 18 mmHg); % By weight] can be further purified by precision distillation or column chromatography. Such compositions consist mainly of acyclic alcohols having 6 carbon atoms, especially primary alcohols, and specifically comprise mainly about 30-55% by weight of trans-2-hexenol,
It is composed of about 15 to 50% by weight of cis-3-hexenol and about 15 to 30% by weight of n-hexanol, but if trans-2-hexenol is contained, the other components and the content ratio of each component are particularly limited. It is not something to be done. For example, in addition to the above components, a minor component such as an isomer of an acyclic alcohol having 6 carbon atoms such as cis-2-hexenol or trans-3-hexenol may be contained.

The microorganism used in the production method of the present invention may be any microorganism as long as it belongs to Candida boidini and selectively oxidizes trans-2-hexenol. For example, Candida boidini SA051 Strains. As described above, this Candida boidini SA051 strain was obtained from Agric. Biol. Chem., 51 (8) pp. 2177-
2184 (1987), a known mutant strain of Candida boidini, which is stored at the Faculty of Agriculture, Kyoto University. The present inventors have deposited this Candida boidini SA051 strain internationally on November 15, 1994 as FERM BP-4893 in the Institute of Biotechnology and Industrial Technology, National Institute of Advanced Industrial Science and Technology.

[0013] The Candida boidini SA051 strain finally used is one that has been cultured using a medium containing methanol alone or a mixture of methanol and glycerin as a carbon source. In such a case, in addition to methanol and glycerin, cultivation may be carried out with a substance that promotes the growth of cells such as ethanol, glucose, xylose, and mannitol. However, the cells cultivated in a medium containing no methanol, such as cells cultivated in a medium containing only glucose as a carbon source or a processed product thereof, when used directly in the production method of the present invention, the property of oxidizing alcohol. Is not sufficiently extracted, so that trans-2-hexenol may not be easily oxidized.

The medium used for culturing and pre-culturing includes, for example, assimilable inorganic nitrogen compounds such as ammonium chloride, ammonium sulfate, ammonium phosphate and urea; potassium phosphate, potassium dihydrogen phosphate, sodium phosphate;
Inorganic salts such as magnesium sulfate, calcium chloride, ferric chloride, ferric sulfate, manganese chloride, manganese sulfate and trace amounts of inorganic salts such as sodium molybdate and potassium iodide; such as disodium ethylenediaminetetraacetate Additives such as organic salts; salts of trace elements such as copper, zinc and cobalt; inorganic acids such as boric acid; and vitamins such as biotin and thiamine hydrochloride are appropriately used for cultivation of general microorganisms. It can be added in a concentration range. A liquid medium is prepared by dissolving these additives and the above carbon source in water. Note that the addition of organic substances such as yeast extract, meat extract, and corn steep liquor is not preferable because the production of alcohol oxidase for producing the intended trans-2-hexenal decreases.

The culture of Candida boidini SA051 strain is carried out by adjusting the pH of the medium to about 4 to 7 at the start of the culture, and thereafter, to about 4 to 7.
~ 6, and under aerobic conditions, at a temperature of about 20-30 ° C,
This is carried out by shaking culture or stirring culture using a rotary shaker, a jar fermenter or the like for 50 to 180 hours. When a jar fermenter is used, an antifoaming agent such as polypropylene glycol may be added to the medium in advance.

In addition, if the operation of adding methanol (and glycerin) is repeated before the methanol (and glycerin) added as a carbon source is completely consumed in the culture, the growth of the bacterial cells is linear. Subsequently, it is possible to increase the amount of dry cells in the culture to a high density of about 100 g / l. The amount of methanol or a mixture of methanol and glycerin is preferably in the range of about 0.5 to 3% by weight based on the total weight of the medium.

The method for producing the trans-2-hexenal-containing composition of the present invention comprises a step of centrifuging the culture solution containing the cells of the Candida boidini SA051 strain obtained as described above as it is or from this culture solution. The suspension obtained by suspending the cells obtained by separation by such means or the processed product thereof, and a trans-2-hexenol-containing composition are mixed, and the cells are shaken or stirred. Alternatively, the treatment is carried out by sufficiently bringing the treated product into contact with the trans-2-hexenol-containing composition.

The concentration of the cells in the reaction solution is preferably in the range of about 5 to 60 g / liter in terms of dry cells. The concentration of the trans-2-hexenol-containing composition in the reaction solution was such that the concentration of trans-2-hexenol contained was about 5 to 150 g /
Liters, and about 10
It is preferably in the range of ~ 300 g / l. The pH of the reaction solution is usually in the range of 6 to 10, preferably in the range of 7 to 9, and any buffer may be used within such a range, and an alkali such as sodium hydroxide may be used. May be added, but it is optimal to use a phosphate buffer. Reaction temperatures are usually in the range of about 4-30 ° C. The reaction is performed under aerobic conditions, but may be performed by injecting air or concentrated oxygen, preferably performed in a closed system under a pure oxygen atmosphere, or performed by injecting pure oxygen while applying pressure. . The reaction time is usually in the range of about 30 minutes to 2 hours, but may vary depending on the composition of the raw material composition and the like. Therefore, the amount of trans-2-hexenal in the composition is sampled and trans- The determination may be made while observing the conversion rate of 2-hexenol to trans-2-hexenal. However, if the reaction is performed for an excessively long time, other alcohols are also oxidized, and unnecessary substances may be generated in the composition.

After completion of the reaction, a known means, for example, an organic solvent such as a hydrocarbon such as n-hexane, an ester such as ethyl acetate, etc.
It can be used as a mixed solvent of more than one kind. ), And concentrated and distilled, so that most of the trans-2-hexenol in the starting material is selectively trans-
The target composition converted to 2-hexenal can be obtained. Since this composition has a strong and preferable green note, it can be used by blending it with a fragrance composition for the purpose of giving a green note. In particular, when the obtained trans-2-hexenal-containing composition is used by being blended with a fragrance composition, the content of trans-2-hexenol remaining in the trans-2-hexenal-containing composition is 25% by weight or less. And more preferably 20% by weight or less.

Further, when the composition obtained by the production method of the present invention is separated into an aldehyde component and an alcohol component by means such as column chromatography, trans-2-hydroxy having a purity of about 90 to 95% as aldehyde component. Hexenal can be obtained. If a trace amount of n-hexanal is removed by such means as further precision distillation,
Further, high-purity trans-2-hexenal can be obtained. The isolated trans-2-hexenal can also be used by blending it in the flavor composition.

The alcohol content obtained by removing the aldehyde component from the composition obtained by the production method of the present invention as described above reduces the content of trans-2-hexenol, which has an unpleasant aroma as a green note. And
It is a component having a preferable green note with an increased content ratio of cis-3-hexenol and n-hexanol having a preferable aroma. This alcohol content can also be used by being blended with the fragrance composition. Further, the alcohol component having a reduced content of trans-2-hexenol is repeatedly brought into contact with the cells of Candida boidini or a processed product thereof based on the production method of the present invention, and the alcohol component remains in the alcohol component. Trans-2-hexenol can be converted to trans-2-hexenal to give a composition having more preferable green notes.

The fragrance composition of the present invention is characterized by containing the trans-2-hexenal-containing composition obtained as described above. The fragrance composition of the present invention comprises a trans
It can be obtained by blending the 2-hexenal-containing composition as it is, or with other commonly used compounded fragrances, that is, synthetic fragrances, natural fragrances, essential oils and the like. The perfume composition of the present invention can be added to foods and drinks, cosmetics, and the like to increase its commercial value.

Trans-2 blended in the fragrance composition of the present invention
As the -hexenal-containing composition, it is preferable to use a composition having a higher content of trans-2-hexenal and a lower content of trans-2-hexenol.Specifically, as a main component, About 20 to 45% by weight of trans-2-hexenal, 0 to 20% by weight of trans-2-hexenol, about 15 to 45% by weight of cis-3-hexenol, and about 15 to 30% by weight of n-hexanol It is preferred to use a composition that does. The content ratio of the trans-2-hexenal-containing composition in the fragrance composition of the present invention is preferably in the range of about 0.01 to 1% by weight.

Further, the fragrance composition of the present invention includes, for example, synthetic fragrances and natural fragrances having a fruity fragrance such as apple, strawberry, orange, grapefruit, and grape; for example, almond oil, neroli oil, orris oil Essential oils such as rose oil, mandarin oil, orange essence, grapefruit essence, and grape essence; for example, absolutes such as Gene Absolute and processed products of essential oils such as terpene-less mandarin oil; Alcohol extract of perfume raw material such as infusion such as tincture and strawberry infusion; alcohols such as ethyl alcohol and octanal; esters such as ethyl butyrate and ethyl isovalerate; fusel oil It can be formulated. The content ratio of these is not particularly limited and can be appropriately changed depending on the use.

The amount of the obtained fragrance composition when blended in foods and drinks, cosmetics and the like depends on the application and is not particularly limited. For example, when blended in beverages, It is preferable to mix the flavor composition in the beverage in an amount of about 0.03 to 1% by weight.

The transformer obtained as described above
Trans-2-hexenal isolated from the 2-hexenal-containing composition can also be used as a fragrance composition by blending it with other compounded fragrances as described above. In this case, the blending ratio of trans-2-hexenal is preferably in the range of about 0.002 to 0.45% by weight based on the flavor composition.

The production method of the present invention can be widely applied to a method for producing trans-2-alkenal by oxidizing trans-2-alkenol in addition to trans-2-hexenol. That is, by contacting the cells of Candida boidini SA051 strain or a treated product thereof, a branched or straight-chain trans-2-alkenol having about 4 to 10 carbon atoms is converted into the corresponding trans-2-alkene. It can be efficiently converted to alkenal. When the trans-2-alkenol is present as a mixture with another alcohol having the same number of carbon atoms, the oxidation rate of the trans-2-alkenol in the mixture is significantly higher than that of the other alcohol, and the Before is oxidized,
Selectively oxidize trans-2-alkenol to trans-
2-Alkenal can be obtained.

[0028]

EXAMPLES Next, the present invention will be described specifically with reference to examples, but the present invention is not limited to these examples. Example 1 Preparation of Cell Suspension I Glucose 20 g, ammonium sulfate 3 g, potassium dihydrogen phosphate 4 g, magnesium sulfate 0.4 g, calcium chloride
10 mg, manganese chloride 2 mg, zinc sulfate 5 mg, biotin 0.05
mg and thiamine hydrochloride (5 mg) were dissolved in ion-exchanged water (1000 ml), and the resulting solution was adjusted to pH 6.0 by adding an 8% by weight aqueous sodium hydroxide solution. Thus, a liquid medium was prepared. Next, 100 ml of the liquid medium was added to a 500 ml foldable Sakaguchi flask with a fold, and the flask was placed in an autoclave and heated at 121 ° C. for 10 minutes for sterilization. Subsequently, the medium in the flask was filled with Candida
Apply Boidini SA051 strain twice using a platinum loop and use a rotary shaker at 140 rpm and 28 ° C under ventilation.
The cells were shake-cultured for 48 hours to obtain a preculture solution.

Next, a liquid medium having the same composition as the above liquid medium except for glucose was prepared and sterilized in the same manner as above, and then 1.5 ml of methanol was added per 100 ml of the liquid medium, and the preculture was added thereto. 5 ml of the solution was added, and the cells were shake-cultured at 140 rpm at 28 ° C. for 65 hours using a rotary shaker. The cells are separated from the obtained culture by centrifugation, washed with a 0.1 M (= mol / l) phosphate buffer solution at pH 7.5, and dried so that the amount of cells is 60 g / l.
Resuspended in 0.1 M phosphate buffer. Thus, the cell suspension I was obtained.

Trans-2-hexenol containing composition I
A commercially available spearmint oil (manufactured by AM TODD) was distilled to obtain a first obtained fraction [distillation temperature: 70 to 80 ° C (13 to 18 mmHg)
, A ratio to spearmint oil: about 10% by weight], and the fraction was again subjected to precision distillation [distillation temperature: 70-80 ° C. (13-80%).
18 mmHg)] to remove contaminants due to azeotropic distillation during the first distillation. Subsequently, silica gel 60 (manufactured by Merck) was used as a stationary phase, and n-hexane: ethyl acetate = 98:
The eluted product was removed by column chromatography using the mixed solution of No. 2 and then eluted through a mixed solution of n-hexane: ethyl acetate = 80: 20 as a mobile phase through the stationary phase to collect the eluted product. By removing n-hexane and ethyl acetate in the fractionated eluate, a purified trans-2-hexenol-containing composition I was obtained (yield based on spearmint oil: 0.08%). This trans-2-hexenol-containing composition I comprises trans-2-hexenol 32.
It contained 6% by weight, 45.3% by weight of cis-3-hexenol, 16.2% by weight of n-hexanol and other acyclic alcohol isomers having 6 carbon atoms.

Trans-2-hexenal-containing composition I
The above cell suspension I (10 ml) and the trans-2-hexenol-containing composition I (0.3 g) were placed in a 100-ml Erlenmeyer flask, the inside of the flask was replaced with pure oxygen, and the flask was sealed. And shaken at 170 rpm for 1 hour at 25 ° C.

After the obtained reaction solution was extracted with 10 ml of n-hexane, n-hexane was removed using a KD concentrator and a rotary evaporator. Thus, trans-2-hexenal 20.8% by weight, trans-2-hexenol 8.0% by weight, cis-3-hexenol 36.3% by weight, n-
Hexanol 15.8% by weight, n-hexanal 1.2% by weight
A trans-2-hexenal-containing composition I 0.
28 g (93.3% yield) were obtained. This trans-2-hexenal-containing composition I had a strong and favorable green note.

Example 2 Preparation of Cell Culture Solution II Glucose 1 was placed in a 70-liter jar fermenter.
20 g, ammonium chloride 305.2 g, potassium dihydrogen phosphate 112.4 g, magnesium sulfate 23.6 g, calcium chloride
2.2 g, ferric chloride 1.5 g, manganese sulfate 0.68 g, disodium ethylenediaminetetraacetate 18 g, zinc sulfate 0.88
g, copper sulfate 0.16 g, cobalt chloride 0.11 g, sodium molybdate 0.1 g, boric acid 0.16 g, potassium iodide 24 m
g, 2 mg of biotin, 200 mg of thiamine hydrochloride and 1.6 g of polypropylene glycol were added and dissolved by adding 40 liters of ion-exchanged water. The resulting solution was adjusted to pH 6.0 by adding an 8% by weight aqueous sodium hydroxide solution. Thus, a liquid medium was obtained. This liquid medium is grown at 121 ° C for 10
It was sterilized by heating for minutes. Subsequently, 800 ml of a culture solution of Candida boidini SA051 strain precultured in a liquid medium having the same composition as the liquid medium in the same manner as the preculture method of Example 1 in the liquid medium in the jar fermenter. The mixture was added and agitated culture was performed at 28 rpm at 200 rpm under aeration at 20 liter / min.

Sixteen hours after the start of the culture, the pH was adjusted to 5.0 by adding an 8% by weight aqueous sodium hydroxide solution, and then 400 ml of methanol was added to continue the culture while maintaining the pH at 5.0. Before the added methanol was completely digested, the addition of 1.2% by weight of methanol to the culture was repeated, and the culture was performed for a total of 105 hours from the start of the culture. g / liter. The time-dependent changes in the concentration of methanol and the amount of bacterial cells were examined. The result is shown in FIG. The cells are separated from the obtained culture by centrifugation, washed with 0.1 M phosphate buffer at pH 7.5, and then re-dissolved in 0.1 M phosphate buffer so that the dry cell amount becomes 60 g / l. Suspended. Thus, a cell suspension II was obtained.

The composition of trans-2-hexenol-containing composition II
A commercially available spearmint oil (manufactured by AM TODD) was distilled to obtain a first obtained fraction [distillation temperature: 70 to 80 ° C (13 to 18 mmHg)
, Relative to spearmint oil: about 10% by weight], and the fraction was purified in the same manner as in the preparation of trans-2-hexenol-containing composition I of Example 1.
Thus, trans-2 comprising 50.8% by weight of trans-2-hexenol, 19.9% by weight of cis-3-hexenol, 25.8% by weight of n-hexanol and other acyclic alcohol isomers having 6 carbon atoms. -Hexenol-containing composition II was obtained (yield based on spearmint oil: 0.08
%).

The trans-2-hexenal-containing composition II
Preparation 3 liters of the above-mentioned cell suspension II and 150 g of the trans-2-hexenol-containing composition II were placed in a jar having a capacity of 5 liters, and pure oxygen was supplied at a rate of 150 ml / min. Stirred at 25 ° C. for 1 hour. The resulting reaction solution was subjected to steam distillation at normal pressure to obtain a 600 ml fraction. 30 g of common salt was added to this fraction and allowed to stand, and the upper layer was separated. Thus, trans-2-hexenal 31.5% by weight, trans-2-hexenol 19.1% by weight, cis-3-hexenol 1
141 g of trans-2-hexenal-containing composition II containing 6.3% by weight, 26.0% by weight of n-hexanol and 1.6% by weight of n-hexanal was obtained (yield: 94.0%). This trans-2-hexenal-containing composition II had a strong and favorable green note.

Column chromatography using 700 g of silica gel 60 (manufactured by Merck) as a stationary phase for isolating trans-2-hexenal and 2 liters of a mixed solution of n-hexane: ethyl acetate = 98: 2 as a mobile phase was carried out.
The aldehyde component contained in the trans-2-hexenal-containing composition II was separated together with the solvent. The solvent was removed by distilling the solution containing the fractionated aldehyde, and 93.0% by weight of trans-2-hexenal and n
An aldehyde content of 45.0 containing 4.7% by weight of hexanal
g (30.0% yield) was obtained. This component had a very strong and favorable green note. Furthermore, precision distillation of this component [distillation temperature: 45 to 55 ° C (13 to 18 mmHg)
As a result, 40.0 g of trans-2-hexenal having a purity of 97% was obtained (yield: 26.7%).

The above transformer was obtained by column chromatography using 700 g of silica gel 60 (manufactured by Merck) as the stationary phase and 2 liters of a mixed solution of n-hexane: ethyl acetate = 80: 20 as the mobile phase. The alcohol content contained in the component obtained by removing the aldehyde component from the 2-hexenal-containing composition II was separated together with the solvent. After distilling the solvent containing the fractionated alcohol to remove the solvent, 30.8% by weight of trans-2-hexenol, cis-
85.6 g (yield: 57.1%) of an alcohol containing 2hex% of 3-hexenol and 42.0% by weight of n-hexanol was obtained. This component had an improved green note over the raw trans-2-hexenol containing composition II.

Example 3 Preparation of Apple Flavor No. 1 Trans-2 obtained in Example 1 according to the recipe shown in Table 1.
-Apple flavor No. 1 blended with hexenal-containing composition I was prepared. Preparation of Apple Flavor No. 2 (Reference Example) According to the formulation shown in Table 1, Apple Flavor No. 2 containing trans-2-hexenal having a purity of 97% obtained in Example 2 was prepared. Preparation of Apple Flavor No. 3 (Reference Example) According to the formulation shown in Table 1, the trans
Apple flavor No. 3 was prepared by blending an alcohol component obtained by removing an aldehyde component from the 2-hexenal-containing composition II. Preparation of Apple Flavor No. 4 (Comparative Example) According to the formulation shown in Table 1, an apple flavor containing trans-2-hexenol-containing composition II used as a starting material in Example 2 was prepared.

[0040]

[Table 1]

The obtained apple flavors No. 1 to No. 4
Is added to commercially available fruitless apple juice and 10% fruit juice apple juice at a ratio of 500 ppm, respectively, to produce a specialized flavorist (the creator of food flavors. Motoichi Indo "Practical knowledge of flavors" (March 1975) 25) Toyo Keizai Shimpo 205
The fragrance was evaluated according to the method of the present invention, and the apple juice to which the trans-2-hexenal-containing composition I obtained by the production method of the present invention was added and the apple flavor No. 1 was added was all of Comparative Example. Than the apple juice to which apple flavor No. 4 containing the starting material trans-2-hexenol-containing composition II was added,
The characteristics of the natural apple had a very pronounced aroma.
As reference examples, apple juice containing apple flavor No. 2 containing trans-2-hexenal and apple juice containing apple flavor No. 3 containing alcohol except for aldehyde were also used as comparative examples. Compared to apple juice to which Apple Flavor No. 4 was added, the characteristics of natural apples had a pronounced aroma.

Example 4 Preparation of strawberry flavor According to the following formulation, a strawberry flavor containing the trans-2-hexenal-containing composition II obtained in Example 2 was prepared. The resulting strawberry flavor had an aroma with very distinctive characteristics of natural strawberry juice. (Prescription) Strawberry infusion: 99.9 parts by weight (Strawberry alcohol extract, manufactured by Takasago International Corporation) Trans-2-hexenal-containing composition II: 0.1 parts by weight

Example 5 Preparation of flavor for orange juice According to the following formulation, a flavor for orange juice containing the composition II containing trans-2-hexenal obtained in Example 2 was prepared. The resulting orange juice flavor had a very pronounced aroma characterized by the characteristics of natural orange juice. (Prescription) Natural orange essence: 99.9 parts by weight (essential oil, manufactured by Takasago International Corporation) Trans-2-hexenal-containing composition II: 0.1 parts by weight

Example 6 Preparation of Grapefruit Flavor A grapefruit flavor containing the trans-2-hexenal-containing composition II obtained in Example 2 was prepared according to the following formulation. The grapefruit flavor obtained had an aroma with very distinctive characteristics of natural grapefruit juice. (Prescription) Natural grapefruit essence: 99.5 parts by weight (essential oil, manufactured by Takasago International Corporation) Natural octanal: 0.2 parts by weight (octanal derived from natural products, manufactured by Takasago International Corporation) Natural nootkatone: 0.2 parts by weight (natural products) Derived nootkatone, manufactured by Takasago International Corporation Co., Ltd. Trans-2-hexenal-containing composition II: 0.1 part by weight

Example 7 Preparation of Grape Flavor A grape flavor containing the trans-2-hexenal-containing composition II obtained in Example 2 was prepared according to the following formulation. The grape flavor obtained had an aroma very distinctive of natural grape juice. (Prescription) Natural grape essence: 99.3 parts by weight (essential oil, manufactured by Takasago International Corporation) Ethyl natural butyrate: 0.5 parts by weight (ethyl acetate derived from natural products, manufactured by Takasago International Corporation) Terpeneless mandarin oil: 0.1 parts by weight ( Essential oil excluding terpenes, manufactured by Takasago International Corporation. Composition containing trans-2-hexenal II: 0.1 part by weight

Reference Example Trans-2-alkenol and other alcohol mixtures having the same number of carbon atoms were prepared by using Candida boidini SA.
The selective oxidation of trans-2-alkenol was examined by contact with the cells of the strain 051.

Experiment No. 1 was treated with 0.1 M so that the dry cell mass was 33 g / l.
3 ml of a cell suspension III prepared in the same manner as the cell suspension I prepared in Example 1 except that the cell suspension was suspended in a phosphate buffer.
And a mixture of trans-2-hexenol and another alcohol having 6 carbon atoms (trans-2-hexenol,
n-hexanol, cis-2-hexenol, trans-3-
90 mg of a mixture containing the same weights of hexenol and cis-3-hexenol), and reacted at 27 ° C. to form an aldehyde (trans-2) corresponding to each alcohol.
-Hexenal, n-hexanal, cis-2-hexenal, trans-3-hexenal, and cis-3-hexenal) in the reaction solution were examined over time. The result is shown in FIG.

Experiment No. 2 Except that trans-2-heptenol and another alcohol mixture having 7 carbon atoms (a mixture containing the same weight of trans-2-heptenol and n-heptanol) were used as the alcohol mixture. The reaction was carried out in the same manner as in Experiment No. 1, and the change over time in the concentration of the aldehyde (trans-2-heptenal and n-heptanal) corresponding to each alcohol in the reaction solution was examined. The result is shown in FIG.

Experiment No. 3 As the alcohol mixture, trans-2-octenol and another alcohol mixture having 8 carbon atoms (a mixture containing the same weight of trans-2-octenol and n-octanol) were used. The reaction was carried out in the same manner as in Experiment No. 1, and the change over time in the content of aldehydes (trans-2-octenal and n-octanal) corresponding to each alcohol in the reaction solution was examined. FIG. 4 shows the results. From these experiments, it was confirmed that in all cases, trans-2-alkenol was selectively oxidized remarkably quickly to produce trans-2-alkenal.

[0050]

According to the production method of the present invention, transformer-2
-Trans-2-hexenol having an unpleasant odor as a green note in a hexenol-containing composition is selectively oxidized to be converted to trans-2-hexenal, whereby trans having a preferred green note is obtained.
A 2-hexenal-containing composition can be obtained. Also, trans-2-hexenal having a preferable green note can be obtained. Since the composition obtained by the method of the present invention has a preferable green note, the perfume composition containing the composition can be added to foods and drinks and cosmetics to enhance its commercial value.

[Brief description of the drawings]

FIG. 1 is a graph showing a time-dependent change in the concentration of methanol and the amount of cells in Example 2;

FIG. 2 is a graph showing a change over time in the concentration of an aldehyde (such as trans-2-hexenal) in a reaction solution.

FIG. 3 is a graph showing a change over time in the concentration of an aldehyde (such as trans-2-heptenal) in a reaction solution.

FIG. 4 is a graph showing a change over time in the concentration of an aldehyde (such as trans-2-octenal) in a reaction solution.

──────────────────────────────────────────────────続 き Continued on the front page (58) Field surveyed (Int. Cl. ⁷ , DB name) C12P 7/24 C11B 9/00-9/02 CA (STN) REGISTRY (STN)

Claims (6)

(57) [Claims] 1. A method for converting trans-2-hexenol to trans-2-hexenal by bringing a cell of Candida boidinii or a processed product thereof into contact with a composition containing trans-2-hexenol. A method for producing a trans-2-hexenal-containing composition, comprising: 2. The trans-2-hexenal according to claim 1, wherein the trans-2-hexenol-containing composition contains trans-2-hexenol and another acyclic alcohol having 6 carbon atoms. A method for producing the composition. 3. The method for producing a trans-2-hexenal-containing composition according to claim 1, wherein the trans-2-hexenol-containing composition is obtained by distilling mint oil. 4. Candida boidini (Candida bo)
The trans-2 according to any one of claims 1 to 3, wherein the cells of idinii) or a processed product thereof are cultured in a medium containing methanol or a mixture of methanol and glycerin as a carbon source. -A method for producing a hexenal-containing composition. 5. A trans-2-hexenal, which is isolated from the trans-2-hexenal-containing composition obtained by the production method according to any one of claims 1 to 4. Method for producing hexenal. 6. A fragrance composition comprising a trans-2-hexenal-containing composition obtained by the production method according to claim 1. Description: