JP3532682B2 - Natural vanilla flavor and method for producing the same - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to vanilla blue pod or ground product, certain yeasts, bacteria, natural vanilla flavor, characterized in that the action of microorganisms selected from among fungi, and to a method for producing .

[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, the present inventors have found that vanilla
By applying a specific microorganism selected from specific yeasts, bacteria and fungi, which has a function of decomposing the precursor of the flavor component contained in vanilla, to the blue pod or its crushed product, a short time of several hours It was found that the vanilla flavor can be easily obtained with. 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.

Namely, the present invention is easy to produce vanilla-like aroma of vanilla blue pod or ground product, and the vanilla flavor production method is rapid, subtly modulated, differentiated natural It is intended to provide vanilla flavoring.

[0007]

To achieve the above object, according to an aspect of the present invention, the vanilla green pods or ground product, Hansenula, that the action of microorganisms selected from microorganisms belonging to the genus Saccharomyces or Bacillus The method is for producing a characteristic natural vanilla flavor. Vanilla here
The blue pod refers to the blue pod itself of vanilla and the whole part 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.

[0009]

The microorganisms used in the present invention include Saccharomyces and Hansenu.
la), a Bacillus (Bacillus).

[0011]

[0012]

[0013] In the product invention, vanilla blue pod or ground product is used. As a method of obtaining a vanilla blue pods pulverized material after the addition of direct or water vanilla blue pod juice - like way to ground by such a method, such as minced by blade knife or the like - Sa - mixer. 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.

[0014] Vanilla blue pod or ground product, as a method for applying a microorganism having an activity of decomposing a precursor of perfume ingredients contained in vanilla, malt or potato extract medium, peptone medium, such as mineral salts medium precultured microorganisms in a suitable medium, after sufficient growth, a method of performing shaking culture in a heterogeneous system with the addition of vanilla blue pod or ground product, a microorganism and vanilla blue pod or into the medium A method of shaking culture in a heterogeneous system in which the pulverized product is added at the same time can be mentioned. Also, in order to increase the yield of vanilla flavor,
It is also possible to add a surfactant to the medium.

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 to act microorganisms vanilla blue pod or ground product, gas chromatography - can be determined while confirming the generation of aroma components using. Although it varies depending on the type of microorganisms used, the composition of the medium, the culture temperature, and the scent of the vanilla flavor to be produced, it is usually preferably 30 minutes to 20 hours. 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 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 condition 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 2 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. The results are shown in Table 1.
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 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 front page (51) Int.Cl. ⁷ Identification code FI // (C12P 7/22 C12R 1:78 C12R 1:78) 1: 865 (C12P 7/22 C12R 1:07 C12R 1: 865 ) (C12P 7/22 C12R 1:07) (72) Inventor Makoto Takahashi 335 Kariyajuku, Nakahara-ku, Kawasaki City, Kanagawa Prefecture Hasegawa Koryo Co., Ltd. Kawasaki Laboratory (72) Minor Iwamoto 335, Kariyajuku, Nakahara-ku, Kawasaki City, Kanagawa Prefecture Address Hasegawa Koryo Co., Ltd. Kawasaki Laboratory (56) Reference JP 58-43757 (JP, A) JP 5-244965 (JP, A) JP 7-87987 (JP, A) JP 3-30683 (JP, A) Lebensm Wiss Technol (1990), Vol. 23, No. 1, p. 1-3 Curr Microbiol (1987), Vol. 16, No. 2, p. 69-73 Fragrance (1993), No. 180, p. 113- 123 (58) Fields investigated (Int.Cl. ⁷ , DB name) C11B 9/00 A23L 1/221 C12P 1/00-39/00 BIOSIS / WPI (DIALOG) JSTPlus (STN)

Claims (2)

(57) [Claims] To 1. A vanilla blue pod or ground product, Hansenula, manufacturing method of natural vanilla flavor, characterized in that the action of microorganisms selected from microorganisms belonging to the genus Saccharomyces or Bacillus. 2. A natural vanilla flavor obtained by the method of claim 1.