TWI670016B - Use of yeast extract - Google Patents

Abstract

Although glutathione is known to have various cosmetic effects such as a high melanin production inhibitory effect and antioxidant effect, it is still classified as a pharmaceutical product in Japan and cannot be used as a food. However, especially in recent years, such functions are required in foods.

A mouse B16 melanoma cell was used to confirm the effect of inhibiting melanin production on a glutathione-containing yeast extract extracted from a yeast that has been edible and recognized for its safety. The results confirmed that the same concentration of glutathione Compared with peptide simple substance, the melanin production inhibitory effect can be more than doubled.

Description

Uses of yeast extract

The invention provides the use of yeast with edible experience and recognized safety, extracting a yeast extract after bacterial cell culture, and using the obtained yeast extract and a melanin production inhibitor of glutathione.

It is important for women to counteract the deterioration of skin symptoms such as pigmentation, wrinkles, and sagging caused by age, stress, and exposure to ultraviolet rays, and to maintain beautiful skin. Therefore, I have sought various means for preventing or improving the deterioration of skin symptoms, and various studies have been completed so far. In particular, it is known that pigmentation, spots, pigmentation after sun exposure, and the like are caused by the activation of pigment cells (melanocytes) existing in the skin, which significantly increases melanin production. As one of the materials for preventing or improving the deterioration of the skin symptoms involved in these pigmentation, glutathione is known.

Glutathione refers to a tripeptide that is a bond of three amino acids, glutamic acid, cysteine, and glycine. It has the antioxidant function of protecting cells from reactive oxygen or peroxidants in cells Important effects in living organisms, such as the action of substances, or the detoxification of foreign matter that is excreted out of cells. In the skin, live Sexual oxygen is also the cause of melanin pigmentation, and it is thought that pigmentation and pigmentation can be suppressed by the antioxidant effect of glutathione.

[Prior technical literature]

[Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open No. 2002-29920

Although glutathione is known to have various cosmetic effects such as a high melanin production inhibitory effect and antioxidant effect, it is still classified as a pharmaceutical in Japan and cannot be used as a food.

An object of the present invention is to provide a melanin production inhibitor having high safety and excellent whitening effect as a food or a pharmaceutical.

In view of this fact, I have been working hard to repeat the research and found that: Torula yeast, which has experience in food consumption and is recognized as safe, extracts yeast extract after bacterial culture, and the resulting glutathione-rich yeast extract The compound has a higher melanin production inhibitory effect than that of glutathione alone, and completed the present invention.

That is, the present invention provides: (1) a melanin production inhibitor containing yeast extract and glutathione; (2) a food and drink product containing a melanin production inhibitor such as (1); (3) a cosmetic product containing (1) ), A melanin production inhibitor; (4) A pharmaceutical product containing a melanin production inhibitor such as (1).

A mouse B16 melanoma cell was used to confirm the effect of suppressing melanin production on a glutathione-containing yeast extract extracted from a edible yeast that has experience in food consumption and is recognized for its safety. Compared to glycine alone, it has more than two times better melanin production inhibitory effect.

1‧‧‧control group

2‧‧‧glutathione

3‧‧‧ Invention of the case (yeast extract containing glutathione)

4‧‧‧ Positive control (sodium lactate)

Figure 1 shows the melanin production inhibitory effect.

The yeast used in the present invention can be used without particular limitation. For example, when extracted from a glutathione-containing yeast, it is preferable to obtain a glutathione-containing yeast extract containing a yeast extract and glutathione. A particularly good yeast line is Candida utilis, commonly known as "round yeast". The culture form of the yeast is not particularly limited, and generally, either batch culture or continuous culture is used. The medium can also be used by ordinary users. For example, as the carbon source, glucose, acetic acid, ethanol, glycerin, molasses, sulfurous acid pulp waste liquid, and the like can be used. As the nitrogen source, urea, ammonia, ammonium sulfate, Ammonium chloride, nitrate, etc. As for the sources of phosphoric acid, potassium, and magnesium, general industrial raw materials such as superphosphate lime, ammonium phosphate, potassium chloride, potassium hydroxide, magnesium sulfate, and magnesium chloride are also preferred. In addition, inorganic salts such as zinc, copper, manganese, and iron ions are generally added. In addition, vitamins, amino acids, and nucleic acid-related substances are also added. Organic substances such as casein, yeast extract, meat extract, coriander can still be added. The culture temperature is 21 to 37 ° C, preferably 25 to 34 ° C; the pH is 3.0 to 8.0, and preferably 3.5 to 7.0. Since the productivity of amino acids or nucleic acids varies depending on the culture conditions, the conditions are set in accordance with the product specifications of the target yeast extract.

After the bacterial cells are cultured, the yeast cells are extracted. The extraction method is not particularly limited, and generally, it can be performed by a self-digestion method, a hot water extraction method, an enzyme extraction method, an acid or alkali extraction method, or a combination thereof.

When extraction is performed by autodigestion, for example, it is stirred at 55 ° C for 4 hours. For example, the enzyme extraction method uses cell wall to dissolve enzymes or proteases for stirring extraction. For example, the acid extraction method is performed after the acid is adjusted to be acidic with sulfuric acid or the like. In the case of alkali extraction, extraction is performed after adjustment to alkalinity. After self-digestion (for example, stirring at 55 ° C for 4 hours), a combination of enzyme extraction and the like may be used.

After extraction, the yeast residue is separated by centrifugation, and concentrated and then freeze-dried or hot air dried to obtain a yeast extract containing glutathione. The glutathione contained in the yeast extract may be any of reduced glutathione, oxidized glutathione, and mixtures thereof. The content of glutathione in the yeast extract may be any content as long as the glutathione is contained in the yeast extract. For example, it is a yeast extract containing glutathione of 3 mass% or more, Preferably it is 10 mass% or more. More preferably, it is a yeast extract which contains 15 mass% or more of glutathione in a dry yeast extract. In order to maximize the content of glutathione in yeast extract Above 15%, the higher the content of glutathione in the cultured yeast, the better. According to this, the efficiency of extracting the yeast extract according to the aforementioned extraction method is better. As a method for increasing the content of glutathione in yeast, a known method is preferred. For example, there is a method of adding zinc ions to a culture medium (Japanese Patent Laid-Open No. 2000-279164); a method for obtaining a glutathione-rich yeast strain using cadmium resistance and macrolide resistance as indicators (Japan) (Japanese Laid-Open Patent Publication No. 2006-42637, Japanese Laid-Open Patent Publication No. 2006-42638). As described in Japanese Patent Application Laid-Open No. 2011-103789, it can also be obtained by extraction of mutant yeast cells. Alternatively, a commercially available yeast extract may be used. For example, there are "Hithionextract" YH-15 and YH-D18 manufactured by KOHJIN Life Sciences.

The method for measuring the glutathione concentration was performed by a DTNB-HPLC method (Journal of Chromatography, 194 (1980) 424-428). The glutathione content in this case refers to the total glutathione content of a combination of reduced and oxidized glutathione.

In the present invention, due to the synergistic effect of the ingredients in the yeast extract and glutathione, it is also possible to add glutathione to a yeast extract that has a low content of glutathione or does not contain glutathione Made of yeast extract containing glutathione.

The present invention provides a food or drink, a cosmetic, a skin external preparation, or a medicinal product characterized by containing the above-mentioned glutathione-rich yeast extract of the present invention.

The food and drink in the present invention refers to a person who orally ingests foods, drinks, hobby products, and supplements with the above composition blended. The form is not particularly limited, and can be used: those who can make staple foods such as bread and noodles; It can be made into non-staple food such as cheese, ham, viennese sausage, processed fish and shellfish; fruit juice, carbonated beverage, milk beverage and other beverages; biscuits, cake, jelly, pudding, candy, yogurt, etc. Moreover, the form of a supplement is not specifically limited, The form of a tablet, a capsule, a soft capsule, and a nutrition drink can also be used.

The blending amount of the above composition in food and drink is not particularly limited as long as it has a melanin production suppressing effect. For example, a yeast extract containing glutathione may contain 10 pg to 20 g with respect to 100 g of food and drink. Among them, it is preferably 100 pg to 2 g, and more preferably 10 ng to 500 mg.

The cosmetic of the present invention refers to a lotion, an emulsion, a foundation, a lipstick, and the like mixed with the above composition.

The blending amount of the composition is not particularly limited, and for example, it may contain 10 pg to 20 g with respect to 100 g of the mass of the composition. Among them, it is preferably 100 pg to 2 g, and more preferably 10 ng to 500 mg.

The present invention can also be used as a pharmaceutical for external use on skin. Although it means a shape of a lotion, an emulsion, an ointment, a spray, a patch (material) and the like in which the above composition is blended, the form is not particularly limited as long as the effect of the purpose of the present invention can be exhibited, it may be any form.

The blending amount of the composition in the pharmaceutical is not particularly limited. For example, a yeast extract rich in glutathione may be 10 pg to 20 g with respect to the mass of the composition. Among them, it is preferably 100 pg to 2 g, and more preferably 10 ng to 500 mg.

[Example]

The following examples are used to describe the present invention in more detail, but the present invention is not limited by these examples.

(Example 1)

Manufacture of glutathione-containing yeast extract

(Culture of yeast)

Candida utilis CS7529 strain (FERMP-3340) was seed-cultured in an Erlenmeyer flask containing YPD medium (1% yeast extract, 2% polyfluorene, and 2% glucose) in a 30L capacity fermentation tank in 18L It was inoculated with 1 ~ 2% in the medium. The composition of the culture medium was 4% glucose, 0.3% monoammonium phosphate, 0.161% ammonium sulfate, 0.137% potassium chloride, 0.08% magnesium sulfate, 1.6 ppm copper sulfate, 14 ppm iron sulfate, 16 ppm manganese sulfate, and 14 ppm zinc sulfate. The culture conditions were performed using pH 4.0, a culture temperature of 30 ° C, an aeration volume of 1 vvm, and stirring at 600 rpm, and addition of ammonia to control the pH. After 16 hours of bacterial cell culture, the culture solution was recovered and the bacteria were collected by centrifugation to obtain 180 g of moist yeast cells.

(Extraction of yeast extract)

The moist yeast cells after culturing the cells were suspended in distilled water and repeatedly centrifuged to perform washing. After washing, the wet bacteria were suspended in distilled water again, or the freeze-dried or hot air-dried bacteria were suspended in distilled water, and adjusted to the following conditions to extract the extract.

Self-digestion: After adjusting to pH 5.0 with 1N HCl, stir at 55 ° C for 4 hours

Enzyme extraction: After adjusting to pH 7 with 1N NaOH, use cell wall-dissolving enzyme (Tunicase) or protease to perform extraction at 55 ° C for 4 hours with stirring.

Acid extraction: After adjusting to pH 2 or less with 1N sulfuric acid, perform stirring extraction at 65 ° C for 2 minutes

Alkali extraction: After adjusting to pH 13 with 2N NaOH, carry out stirring extraction at 70 ° C for 20 minutes

(Drying of yeast extract)

After extraction, the yeast residue is separated by centrifugation, and the extract is concentrated, and then freeze-dried or hot air dried to obtain a glutathione-containing yeast extract. The reduced glutathione content was 15.51%, and the glutathione content (total glutathione content) was 17.68%.

(Confirmation of melanin production suppressing effect)

The melanin production inhibitory effect was measured by the following procedure.

<Verification of melanin production inhibitory effect>

B16 mouse melanoma cells were seeded on a 6-well plate (2 × 10 ⁴ / well) and cultured 24 hours before (5% CO ₂ , 37 ° C.). D-MEM medium containing 5% fetal bovine serum was used as the culture medium. Thereafter, it was replaced with a fresh medium, and glutathione (Comparative Example 1) and a yeast extract rich in glutathione (Example 1) were added so that the final concentration became 1.0 mM in terms of reduced glutathione. And sodium lactate (Comparative Example 2) which is known to have a melanin production inhibitory effect, so that the final concentration becomes 50 mM. On the fourth day after sowing, the medium was removed, the plate was washed with PBS (phosphate-buffered saline), and then stripped with 0.25% trypsin EDTA, and pellets (cells) were recovered by centrifugation (10,000 rpm, 3 min). After the recovered pellets were washed twice with PBS, the cell suspension was adjusted with 1 mL of PBS, and 0.8 mL was used to measure the amount of melanin, and 0.2 mL was used to quantify the protein.

The recovered pellets used for measuring the amount of melanin were dissolved in 150 μL of 1N NaOH (100 ° C., 10 minutes), and the absorbance at 405 nm was measured using a microplate reader. The pellets used for protein recovery and quantification were quantified by the BCA method. The water-added person was used as a control group. When the control group was set to 100%, the amount of melanin per unit protein of the cells cultured in Comparative Examples 1 and 2 was added as the melanin production rate.

The results of Example 1 are shown in FIG. 1. As shown in FIG. 1, when glutathione is alone (Comparative Example 1), the melanin production inhibition rate is about 40% compared with the control group. In contrast, the glutathione-rich yeast extract of the present invention is The melanin production inhibitory rate was about 85% compared with the control group, and it was judged that the melanin production inhibitory effect was twice or more than that of glutathione alone. It is considered that this is because the components other than glutathione in the glutathione and the yeast extract exhibit a multiplier effect. Therefore, the glutathione-enriched yeast extract of the present invention can be used as a food, and can exhibit an excellent melanin production inhibitory effect, and is useful as a melanin production inhibitor.

Claims (1)

A use of a yeast extract is to use a yeast extract to produce a melanin production inhibitor. The melanin production inhibitor contains a yeast extract and glutathione, and the yeast extract contains 15% by mass or more of glutathione.