JP4655295B2 - Production method of aromatic components derived from mushrooms - Google Patents

Description

  The present invention relates to a method for producing an aroma component derived from mushrooms, and more particularly to a method for efficiently producing an aroma component containing cinnamaldehyde and / or benzaldehyde by culturing a seedling of Grifola galgal.

  Cinnamaldehyde and benzaldehyde are widely used as fragrances and as raw materials for other fragrances and medicines. Many of these have been synthesized, but if they can be efficiently produced in a form derived from natural products, their utility value will be greatly increased in terms of safety.

Conventionally, there are a plurality of proposals for a method for producing an aromatic component derived from a mushroom that is a natural product (see, for example, Patent Documents 1 to 3). Some of these contain benzaldehyde as an aromatic component. However, these conventional methods have a problem that the production efficiency of cinnamaldehyde and benzaldehyde is extremely low.
Japanese Unexamined Patent Publication No. 64-16567 JP-A-6-67 JP 2004-337018 A

  The problem to be solved by the present invention is to provide a method for producing an aromatic component derived from a mushroom which is a natural product, which can efficiently produce cinnamaldehyde and benzaldehyde.

  The present invention that solves the above-mentioned problems is a method of inoculating a culture medium with an inoculum of Grifola galgal, culturing and growing a mycelium, and then aroma containing cinnamaldehyde and / or benzaldehyde from the culture. The present invention relates to a method for producing an aromatic component derived from a mushroom, characterized by separating the components.

  In the present invention, Grifola galgal (hereinafter simply referred to as galgal) is used as a mushroom. Gargar is a mushroom that grows naturally in the Patagonia region of Chile, South America, and its fruiting body is used for food, and has a stronger fragrance compared to Grifola frontosa (Japanese name is Maitake), which is a mushroom of the same Glyfora genus .

  In the present invention, the culture medium is inoculated with a galgal inoculum and cultured to grow mycelium. The medium may be a solid medium or a liquid medium, but a liquid medium is preferred in order to promote efficient production of aroma components and to separate and recover the aroma components from the culture. As the medium itself, a general mushroom medium can be applied. For example, a glucose peptone liquid medium can be applied as the liquid medium. As will be described in detail later, when an inoculum is inoculated into a medium, cultured, and mycelium is grown, an aromatic component mainly composed of cinnamaldehyde and / or benzaldehyde is produced, depending on the time. In order to promote the production of these aromatic components more efficiently, it is also effective to add a precursor of these aromatic components to the medium in advance, for example, phenylalanine in the case of benzaldehyde.

  The pH of the medium, the culture temperature, and the like can also follow general mushroom culture conditions. The pH of the medium can be 3.5 to 8.0, and the culture temperature can be 10 to 30 ° C., as described above. In order to promote the production of aroma components more efficiently, when a liquid medium is used as the medium, the pH is preferably 4.5 to 6.0, and the culture temperature is preferably 20 to 25 ° C.

  As described above, when a culture medium is inoculated with a galgal inoculum and cultured to grow mycelium, a strong aroma component is produced. In general, a fragrance component mainly composed of cinnamaldehyde is produced in the first growth phase of the mycelium, and a fragrance component mainly composed of benzaldehyde is produced in the second growth phase of the mycelium. More specifically, the production amount of cinnamaldehyde suddenly increases in the early growth stage of the mycelium, but this has a peak region, and after the peak region, the production amount suddenly decreases. As if it were replaced with this, the production amount of benzaldehyde suddenly increased in the later stage of growth of the mycelium, but this also has a peak region, and when the peak region is passed, the production amount suddenly decreases.

  In the present invention, the scent component containing cinnamaldehyde and / or benzaldehyde is separated from the culture in the process of inoculating the culture medium with the galgar inoculum, culturing and growing the mycelium. Separation can be performed by extraction using an organic solvent, steam distillation or the like. Cinnamaldehyde and benzaldehyde can also be isolated directly from the culture, usually from aroma components separated from the culture, by column chromatography, molecular distillation or the like. Although it depends on the culture conditions, in general, the production amount of cinnamaldehyde reaches a peak in 10 to 12 days of culture and can be 400 to 700 mg per liter of the culture medium. The production amount of benzaldehyde is 16 days of culture. It becomes a peak region in ˜22 days, and can be 800 to 1500 mg per liter of the medium.

  According to the present invention, cinnamaldehyde and benzaldehyde can be efficiently produced from mushrooms which are natural products.

Test category 1
Native to South America Chilean Patagonia, with Gargano Le that is edible as test bacteria. Glucose / peptone liquid medium (both mass%, glucose 2%, polypeptone 0.3%, yeast extract 0.3%, magnesium sulfate 0.1%, dipotassium phosphate 0.1%, the balance is water, pH 5 0.0) was prepared, and 10 ml each was dispensed into a 100 ml Erlenmeyer flask, and then pasteurized at 121 ° C. for 20 minutes and cooled. Each Erlenmeyer flask is inoculated with one piece of cultured mycelium of the test bacteria punched out with a cork borer with an inner diameter of 5.5 mm, and after standing for 15 days in 5 sections within a culture temperature range of 10 to 30 ° C., the culture is performed. The product was filtered by suction and divided into a mycelium section and a culture liquid section. The mycelium segment was dried at 105 ° C. for 5 hours, the mass was measured, and this was defined as the mass (g) of the mycelium. In addition, after adding ethyl ether to the culture medium and shaking, allowing to stand and separating the layers, the centrifugal solution of the upper ethyl ether layer is subjected to gas chromatography, and the amount of benzaldehyde is measured. The amount was converted to benzaldehyde production (μg) per 1 ml of the medium. The results are shown in FIG.

  FIG. 1 is a graph showing changes in mycelial mass (g) and benzaldehyde production (μg / ml) with respect to culture temperature (° C.) in the present invention. In FIG. 1, a curve 11 connecting triangles indicates the mass (g) of mycelia, and a curve 21 connecting white circles indicates the amount of benzaldehyde produced (μg / ml). As apparent from FIG. 1, the culture temperature can be in the temperature range of 10 to 30 ° C., but is preferably in the temperature range of 20 to 25 ° C.

Test category 2
The cultured mycelium of the test bacteria was cultured in the same manner as in test category 1. However, here, the culture temperature is set to 22 ° C., the liquid medium is left to stand for 11 days in 11 sections within a pH range of 3.0 to 8.0, the mycelium mass (g), the medium The production amount (μg) of benzaldehyde per ml was determined. The results are shown in FIG.

  FIG. 2 is a graph showing changes in the mass (g) of mycelium and the production amount (μg / ml) of benzaldehyde with respect to the pH of the liquid medium in the present invention. In FIG. 2, a curve 12 connecting triangles indicates the mass (g) of mycelia, and a curve 22 connecting white circles indicates the production amount of benzaldehyde (μg / ml). As is apparent from FIG. 2, the pH of the liquid medium can be in the range of 3.0 to 8.0, but is preferably in the range of 4.5 to 6.0.

Test category 3
Into a 3L jar fermenter, add 2.0L of phenylalanine adjusted to 0.02% by mass and adjusted to pH 5.0 in the same liquid medium as in Test Category 1, and pasteurized at 120 ° C for 20 minutes. And cooled. To this, 0.1 L of a preculture solution of a test bacterium similar to that in Test Category 1 was added, followed by aeration and stirring culture at a culture temperature of 22 ° C. for 22 days. During the culture, the culture was appropriately sampled, and the sampled culture was suction filtered to divide it into mycelium sections and culture medium sections. The mycelium segment was dried at 105 ° C. for 5 hours, the mass was measured, and this was defined as the mass (g) of the mycelium. In addition, the centrifuged solution of the culture medium is subjected to high performance liquid chromatography, the amount of cinnamaldehyde and benzaldehyde is measured, and these are converted into cinnamaldehyde production (mg) and benzaldehyde production (mg) per liter of medium. Converted. The results are shown in FIG.

  FIG. 3 is a graph showing changes in mycelial mass (g), benzaldehyde production (mg / L), and cinnamaldehyde production (mg / L) with respect to the number of days of culture (days) in the present invention. In FIG. 3, a curve 13 connecting the triangle marks indicates the mass (g) of the mycelium, a curve 23 connecting the open circle marks indicates the production amount (mg / L) of benzaldehyde, and a curve 33 connecting the black circle marks. Indicates the production amount of cinnamaldehyde (mg / L). As is apparent from FIG. 3, the production amount of cinnamaldehyde is a peak region in the culture period of 10 to 12 days, and is 400 to 550 mg per liter of the culture medium. Therefore, the example of the culture days of 10 to 12 corresponds to a preferred embodiment in the case of producing cinnamaldehyde in the present invention. Moreover, the production amount of benzaldehyde has a peak region at 16 to 22 days of culture, and is 900 to 1050 mg per liter of the medium. Therefore, the example of the culture days of 16 to 22 corresponds to a preferred embodiment when producing benzaldehyde in the present invention.

The graph which shows the change of the mass (g) of a mycelium with respect to culture | cultivation temperature (degreeC) and the production amount (microgram / ml) of a benzaldehyde in this invention. The graph which shows the change of the mass (g) of a mycelium with respect to pH of a liquid culture medium, and the production amount (microgram / ml) of benzaldehyde in this invention. The graph which shows the change of the mass (g) of a mycelium with respect to culture days (days), the production amount (mg / L) of a benzaldehyde, and the production amount (mg / L) of a cinnamaldehyde in this invention.

Explanation of symbols

11, 12, 13 Curve showing the mass of mycelium 21, 22, 23 Curve showing the production amount of benzaldehyde 33 Curve showing the production amount of cinnamaldehyde

Claims (4)

  Mushrooms characterized by separating aroma components containing cinnamaldehyde and / or benzaldehyde from the culture in the process of inoculating the culture medium with an inoculum of Grifola galgal, culturing and growing the mycelium. A method for producing an aromatic component derived from the origin.   The method for producing mushroom-derived aroma components according to claim 1, wherein the medium is a liquid medium.   The method for producing an aroma component derived from a mushroom according to claim 2, wherein the liquid medium is adjusted to pH 4.5 to 6.0. The method for producing an aroma component derived from a mushroom according to any one of claims 1 to 3, wherein the culturing is performed in a temperature range of 20 to 25 ° C.

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