JPH09111285A - Natural vanilla perfume and its production - Google Patents

Abstract

PROBLEM TO BE SOLVED: To produce not only a unified vanilla perfume but also the subject variously differentiated perfume having delicately varying fragrance quality by reacting a vanilla pod or its pulverized material with a specific microorganism. SOLUTION: A vanilla pod or its pulverized material is reacted with a microorganism, having actions on decomposition of a precursor of a perfume ingredient contained in the vanilla and selected from a yeast (e.g. the genus Saccharomyces), a bacterium (e.g. the genus Bacillus) and a mold (e.g. the genus Aspergillus). Furthermore, the objective perfume is preferably obtained by preculturing, e.g. the microorganism in a suitable culture medium, sufficiently proliferating the microorganism, then adding the vanilla pod or its pulverized material thereto, carrying out the shaking culture in a heterogeneous system, completing the culturing and subsequently extracting the prepared supernatant with an organic solvent, etc.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a natural vanilla fragrance characterized by allowing a microorganism selected from specific yeasts, bacteria and fungi to act on a vanilla pod or a crushed product thereof.
And its manufacturing method.

[0002]

2. Description of the Related Art Vanilla fragrances mainly include vanillin, vanillic acid, parahydroxybenzoic acid and parahydroxybenzaldehyde. These aroma components do not exist in the freshly harvested vanilla blue pods of a kind of orchid plant, vanilla, and normally, after the process of curing, the aroma of vanilla is recognized. Become. The vanilla blue pods at the time of harvest contain a large amount of precursors of odor components such as odorless glucovanillin, which is a precursor of vanillin, and decomposition reactions such as cleavage of glucoside bonds due to enzymatic reaction in the curing process. Occurs, the vanilla-like aroma component is released, and aroma is generated. It is known that the method of this curing step differs in each production area, and the difference affects the quality of the vanilla flavor.

Generally, the curing step is carried out by alternating aging and drying until the vanilla blue pods change to a dark brown chocolate color, which requires a long time of 6 to 9 months. It was said that it was difficult to keep the quality of vanilla fragrance stable.

Therefore, in Japanese Patent Publication No. 6-502685, the conversion time from vanilla odorless glucovanillin, which is a precursor of vanillin, to vanillin by treating vanilla blue pods with an enzyme system having glucosidase activity. There has been proposed a method for producing a vanilla flavoring agent which can substantially shorten the above. However, in order to extract and purify the enzyme from the living body, various operation steps are required, and when the enzyme is taken out of the living body, it becomes unstable due to a change in the environment, and therefore, care must be taken in handling the enzyme. In addition, some enzymes are expensive and cannot be used repeatedly, resulting in high cost. Furthermore, the obtained fragrance was a uniform vanilla fragrance, and a differentiated vanilla fragrance having a slightly different fragrance quality was not obtained.

As a result of intensive studies by the present inventors, the present invention is selected from specific yeasts, bacteria, and fungi, which have an action of decomposing the precursor of the flavor component contained in vanilla into vanilla pods or pulverized products thereof. It has been found that vanilla flavor can be easily obtained in a short time of several hours by allowing a specific microorganism to act. Once obtained, the microorganism has the advantage that it can be grown and used repeatedly, and by controlling the type of microorganism used and the culture conditions, some of the liberated vanillin is converted to another compound for secondary metabolism. It also happens, so not only uniform vanilla flavors,
It has been found that a variety of differentiated vanilla fragrances with slightly different scents are obtained.

That is, the present invention makes it easy to produce a vanilla-like aroma component from a vanilla pod or a crushed product thereof, and
A speedy vanilla flavor production method and subtly modulated,
The purpose is to provide a differentiated natural vanilla flavor.

[0007]

In order to achieve the above object, the present invention provides a yeast, a bacterium or a fungus having an action of decomposing a precursor of a perfume ingredient contained in vanilla into a vanilla pod or a pulverized product thereof. A method for producing a natural vanilla flavoring characterized by allowing a microorganism selected from the above to act. The term “vanilla pod” as used herein refers to the vanilla pod itself and the entire portion covered with the pod such as seeds.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The type of vanilla used in the present invention includes vanilla planifol (Vanilla planifol).
ia), Vanilla tahitensis,
It is Vanilla pompona.

The microorganism used in the present invention may be one or more selected from specific yeasts, bacteria and fungi, which have the action of decomposing the precursor of the flavor component contained in vanilla. Mixtures can be used, but fast-growing yeast, bacteria or combinations thereof are preferred.

The yeasts used in the present invention include Saccharomyces and Hansenul.
a), Kluyveromyces (Kluyveromyces), Rodaromyces (Lodderomyces), Pichia (Pichia), Nadsonia, Saccharomycodes
des), Hanseniaspora, Schizosaccharomyces, Endomycopssis, Nematospora
ora), etc. and Endomycetale (Endomycetale
s), Leucospodium (Leucospo)
ridium), those belonging to Ustilaginales, such as Rhodosporidium, Bullera, Sporobomyces
oromyces), such as Sporidiobolus (Sporidiobolus), Sporobolomycetaceae
Belonging to the above (above Ascomycete yeast), Brettanomyces (Brettanomyces), Cryptococcu (Cryptococcu)
s), Kloeckera, Rhodotor
ula), Sterigmatomyces (Sterigmatomyces)
(Above-mentioned non-spore yeast) etc. are mentioned. Saccharomyces and Hansenula, which are easily available and easy to handle, are particularly preferable.

The bacterium used in the present invention is a bacterium having no action of decomposing the precursor of the flavor component contained in vanilla, for example, Staphylococcus aureus (S
taphylococcus aureus) (ATCC12082) and Corynebacterium minuticimas (Cornynebacterium mi
nutissimum) (ATCC23348), and specific examples of the bacterium used in the present invention include, for example, Bacillus, Eschrichia,
Examples thereof include Leuconostoc, but Bacillus, which is easy to handle, is particularly preferable.

Molds used in the present invention include Aspergillus, Rhizopus, Trichoderma and Penicillium.
ium) and the like.

In the present invention, a vanilla pod or a crushed product thereof is used. As a method of obtaining a crushed product of vanilla pods, the vanilla pods may be added directly or after adding water.
Examples thereof include a method of crushing with a mixer and the like, and a method of finely chopping with a knife such as a knife. It is preferable that the seeds and the like are crushed to such an extent that they can be crushed, and crushed more finely so that the degree of contact with the microorganisms is increased as much as possible.

As a method of causing a microorganism having an action of decomposing a precursor of a flavor component contained in vanilla to act on a vanilla pod or a crushed product thereof, malt or potato extract medium, peptone medium, inorganic salt medium or the like is suitable. After preculturing the microorganism in a suitable medium and allowing it to proliferate sufficiently, a method of shaking culture in a heterogeneous system by adding vanilla pod or its crushed product, culturing the microorganism and vanilla pod or its crushed product in the medium A method of shaking culture in a heterogeneous system added at the same time can be mentioned. It is also possible to add a surfactant to the medium in order to increase the yield of vanilla flavor.

After completion of the culture, solid contents such as bacterial cells and vanilla residue are spun down from the culture mixture, the supernatant is extracted with an organic solvent, the organic layer is dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. This gives an essential oil having a vanilla odor.

The time for which the microorganisms act on the vanilla pods or the pulverized products thereof can be determined by confirming the production of aroma components using gas chromatography. Depending on the type of microorganism used, the composition of the medium, the culture temperature, etc., and the scent of the vanilla flavor to be produced, it is usually 30 minutes to 30 minutes.
20 hours is preferred. Within this range, the precursor of the aroma component is sufficiently assimilated, so that the yield of the vanilla flavor is good, and the excessive secondary metabolism of the produced aroma component is suppressed, so that the fragrance of the vanilla flavor is deteriorated. I can't.

The culture temperature of the microorganism is preferably 25 to 40 ° C. Within this range, the microorganisms proliferate smoothly, so that the production reaction of the vanilla-like fragrant substance rapidly progresses, and deterioration of the obtained vanilla fragrance due to heat is suppressed.

[0018]

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to examples.

Example 1 In an Erlenmeyer flask, 20% of malt extract broth medium was inoculated with 1% by weight of Hansenula anomala (isolated strain) and cultured for 24 hours (30 ° C., 100 rpm) to grow sufficiently. To this was added 2.5 g of vanilla blue pods (yellow-green without vanilla odor and pods containing seeds) crushed in advance with a juicer mixer, and the mixture was incubated for 1 hour (30 ° C., 10 ° C.).
0 rpm). The solid content was spun down from the culture mixture, and the supernatant was extracted with the same amount of diethyl ether. The organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure to obtain 5 mg of an essential oil having a vanilla odor.

The gas chromatogram of the essential oil obtained by the method of Example 1 is shown in FIG. The analysis method of gas chromatography described in Examples and Comparative Examples is as follows. Analytical conditions Instrument: Hewlett Packard 5890 column; DB-WAX (30 m x 0.25 nm)
I. D = 0.25 μm carrier gas; 1 ml / min. (He) Extractor inlet pressure =
14 psi column temperature; 100 ° C (1 min.) To 250 ° C
(10 ° C / min.) Injector; 250 ° C, splitless, 1 μl detector; FID, 250 ° C

Example 2 In an Erlenmeyer flask, 16 ml of a 15% by weight aqueous sucrose solution,
Commercial baker's yeast (Saccharomyces cerevisiae) 1.
Add 05g and pre-incubate for 30 minutes (30 ° C, 130rp
m) was performed and allowed to grow well. To this, 5.0 g of vanilla pods crushed in advance with a juicer mixer was added, and the mixture was further cultured for 1 hour (30 ° C, 130 rpm). The solid content was spun down from the culture mixture, and the supernatant was extracted with the same amount of diethyl ether. The organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure to obtain an essential oil having a vanilla odor.

A gas chromatogram of the essential oil obtained by the method of Example 2 is shown in FIG.

Example 3 In an Erlenmeyer flask, 20 ml of SCD broth medium was inoculated with 1% by weight of Bacillus subtilus (IAM1069),
The cells were precultured for 24 hours (30 ° C., 100 rpm) and grown sufficiently. To this, 2.5 g of vanilla pods crushed in advance with a juicer mixer was added, and culturing was carried out for 8 hours (30 ° C., 100 rpm). The solid content was spun down from the culture mixture, and the supernatant was extracted with the same amount of diethyl ether. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain an essential oil having a vanilla odor.

A gas chromatogram of the essential oil obtained by the method of Example 3 is shown in FIG.

Example 4 In an Erlenmeyer flask, 20 ml of SCD broth medium was inoculated with 1% by weight of Bacillus subtilus (IAM1069),
The cells were precultured for 24 hours (30 ° C., 100 rpm) and grown sufficiently. To this, 2.5 g of vanilla pods crushed in advance with a juicer mixer was added, and the mixture was cultured for 48 hours (30 ° C, 100 rpm). The solid content was spun down from the culture mixture, and the supernatant was mixed with an equal volume of diethyl ether.
Extracted with tel. The organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure to obtain an essential oil.

Comparative Example 1 In an Erlenmeyer flask, 20 ml of SCD broth medium was inoculated with 1% by weight of Staphylococcus aureus (ATCC12082), and precultured for 24 hours (30 ° C., 100 rpm).
Was carried out and allowed to grow sufficiently. In advance for this
Add 2.5 g of vanilla pods crushed with a mixer,
Culture was performed for 48 hours (30 ° C., 100 rpm). The solid content was spun down from the culture mixture, and the supernatant was extracted with the same amount of diethyl ether. The organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure, but almost no oil was obtained.

Comparative Example 2 In an Erlenmeyer flask, 20% of SCD broth medium was inoculated with 1% by weight of Corynebacterium minutisimum (ATCC 23348), and precultured for 24 hours (30 ° C., 100 r).
pm) and allowed to grow sufficiently. To this, 2.5 g of vanilla pods crushed in advance with a juicer mixer was added, and the mixture was cultured for 48 hours (30 ° C, 100 rpm). The solid content was spun down from the culture mixture, and the supernatant was extracted with the same amount of diethyl ether. The organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure, but almost no oil was obtained.

Comparative Example 3 A gas chromatogram of commercial vanilla essence is shown in FIG.

Comparative Example 4 A gas chromatogram of synthetic vanillin is shown in FIG.

Sensory test The essential oils of Examples 1 to 4 were subjected to a sensory test by two expert panelists. Using the vanilla essence of Comparative Example 4 as a reference, four items of vanilla odor, fresh sensation, fermentation odor, and difference from vanilla essence were evaluated. Table 1 shows the results
Shown in (+: Yes,-: None for differences from fermentation odor and vanilla essence, ○: excellent, △: comparable, ×: inferior for the other two items)

[0031]

[Table 1]

From the results in Table 1 (the same answer for both panelists), it can be seen that the oils of Examples 1 to 4 are vanilla fragrances which are well differentiated from the commercially available vanilla essence and have excellent sensory characteristics. Examples 1 to 3 which are particularly short-term culture products
The oil had no fermentation odor and had a vanilla odor with a fresh feeling. In addition, from the analysis results of gas chromatography, the oils of Examples 1 to 3 contained phenylethyl alcohol (peak P), vanillyl alcohol (peak B), and guaiacol (peak G) in addition to vanillin (peak V). , Vanilla essence on the market and synthetic vanillin were found to have different fragrances.

[0033]

From the above, the production method of the present invention is
It is clear to provide a subtly modulated and differentiated natural vanilla flavor.

[Brief description of the drawings]

FIG. 1 is a gas chromatogram of the vanilla fragrance of Example 1.

FIG. 2 is a gas chromatogram of the vanilla flavor of Example 2.

FIG. 3 is a gas chromatogram of the vanilla flavor of Example 3.

4 is a gas chromatogram of vanilla essence of Comparative Example 3. FIG.

5 is a gas chromatogram of synthetic vanillin of Comparative Example 4. FIG.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Office reference number FI technical display location // (C12P 7/22 C12R 1:78) (C12P 7/22 C12R 1: 865) (C12P 7 (22) C12R 1:07) (72) Inventor Fumiichi Okabe 5-3 28, Kotobuki-cho, Odawara-shi, Kanagawa, Kanebo Co., Ltd. Cosmetic Research Institute (72) Makoto Takahashi, 335, Kadojuku, Nakahara-ku, Kawasaki-shi, Kanagawa Hasegawa Kaori Co., Ltd., Kawasaki Research Institute (72) Inventor Minoru Iwamoto 335, Kayajuku, Nakahara-ku, Kawasaki, Kanagawa Prefecture Hasegawa Kaori Co., Ltd., Kawasaki Research Institute

Claims (2)

[Claims] 1. A natural vanilla characterized by causing a microorganism selected from yeast, bacteria and fungi, which has an action of decomposing a precursor of a flavor component contained in vanilla, to a vanilla pod or a crushed product thereof. Method for producing fragrance. 2. A natural vanilla flavor obtained by the method of claim 1.