JP6624985B2 - Polyamine-rich yeast - Google Patents

Description

  The present invention relates to a yeast containing a polyamine at a high concentration, and a method for producing the same.

  Polyamine is a general term for a linear aliphatic hydrocarbon having a plurality of primary amino groups and is ubiquitous in the cells of an organism. To date, more than 20 polyamines have been reported. There are mainly three types of polyamines in the human body: putrescine, spermidine and spermine, and the amount decreases at the peak of the 20s. Polyamine is a component necessary for cell division, and it is considered that when the amount of intracellular polyamine decreases, cell turnover decreases and senescence accelerates.

  Therefore, it is expected that various physiological effects can be enjoyed by supplementing polyamine from outside the body. Soybeans, mushrooms, chicken, cheese and the like are known as foods containing a large amount of polyamine. However, since the amount of polyamine contained in ordinary food is not so large, it is not easy to ingest a sufficient amount of polyamine from a daily meal.

  Methods for preparing polyamines from food materials have been proposed. Patent Documents 1 and 2 describe a method of separating and recovering polyamine from milk or milk material such as cheese whey by ultrafiltration or ion exchange treatment. Patent Document 3 describes a method for recovering polyamine from milt by mineral acid treatment and centrifugation. However, the above-mentioned method is low in productivity because the content of polyamine in the raw food material is not large, and is expensive because of requiring complicated steps for purification.

  Patent Document 4 describes that yeast contains a high concentration of polyamine, and a method for separating and purifying a polyamine-containing fraction by extracting yeast cells or a culture solution with an acid solution. . According to Patent Document 4, the conventionally reported polyamine content of yeast is about 13 mg for Saccharomyces cerevisiae, about 23 mg for Saccharomyces diastaticas, and about 103 mg for Candida ulitis per 100 g of dry cells. The total content of putrescine, spermidine, and spermine in 100 g (wet weight) of the bacterial cells calculated from the report of Non-Patent Document 1 is about 17 mg for Saccharomyces cerevisiae, about 30 mg for Candida glabrate, and about 21 mg for Zygosaccharomyces rouxii.

JP-A-6-305956 JP 2001-8663 A JP-A-8-238094 JP-A-10-52291

Bulletin of Gunma Health Science 25: 117-123, 2004

  The present invention provides a yeast containing a polyamine useful as a polyamine source at a high concentration, and a method for producing the same.

  As a result of various studies, the present inventors have found that by culturing yeast in a medium to which a polyamine such as putrescine, spermidine, spermine, and / or a specific type of sodium source has been added, the polyamine content in the yeast cell has increased. Was found to increase. Based on these findings, the present inventors have succeeded in improving the polyamine productivity of yeast and producing a yeast having a significantly higher polyamine content in cells than conventionally known yeasts.

Therefore, the present invention provides a method for producing a polyamine-rich yeast, comprising culturing yeast in a medium to which the following component A and / or component B has been added:
A component One or more polyamines selected from the group consisting of putrescine, spermidine and spermine, in a total of 0.1 mM or more;
B component One or more sodium sources selected from the group consisting of sodium chloride, sodium glutamate, sodium dihydrogen phosphate, citric acid and sodium hydroxide, a sodium salt of citric acid, and sodium malate, having a sodium concentration of 25 mM that's all,
I will provide a.

The present invention also provides a method for improving yeast polyamine productivity, which comprises culturing yeast in a medium to which the following component A and / or component B has been added:
A component One or more polyamines selected from the group consisting of putrescine, spermidine and spermine, in a total of 0.1 mM or more;
B component One or more sodium sources selected from the group consisting of sodium chloride, sodium glutamate, sodium dihydrogen phosphate, citric acid and sodium hydroxide, a sodium salt of citric acid, and sodium malate, having a sodium concentration of 25 mM that's all,
I will provide a.

  Furthermore, the present invention provides a polyamine-rich yeast containing a polyamine in an amount of 150 mg / 100 g cells (in terms of dry matter) or more.

  Further, the present invention provides a polyamine-rich yeast produced by the above production method.

  Further, the present invention provides a method for producing a polyamine, comprising separating the polyamine from the above-described yeast containing high polyamine.

Furthermore, the present invention provides a yeast culture medium containing the following A component and / or B component:
A component One or more polyamines selected from the group consisting of putrescine, spermidine and spermine, in a total of 0.1 mM or more;
B component One or more sodium sources selected from the group consisting of sodium chloride, sodium glutamate, sodium dihydrogen phosphate, citric acid and sodium hydroxide, a sodium salt of citric acid, and sodium malate, having a sodium concentration of 25 mM that's all,
I will provide a.

  According to the present invention, a yeast containing a polyamine at a high concentration can be provided. The polyamine-rich yeast provided by the present invention is useful as a polyamine source to be added to foods and the like.

Amount of polyamine in yeast cells cultured in putrescine-supplemented medium. The amount of polyamine in yeast cells cultured in various sodium-supplemented media. The amount of polyamine in yeast cells cultured in various sodium-supplemented media. The amount of polyamine in yeast cells cultured in an acidic medium supplemented with a sodium source. The amount of polyamine in yeast cells cultured in a medium containing putrescine and a sodium source. For each putrescine concentration, the sodium source concentration was 0, 25, 50, 100 mM from the left. The amount of polyamine in yeast cells cultured in a medium containing trisodium citrate. The amount of polyamine in yeast cells cultured in a medium containing putrescine and trisodium citrate. The amount of polyamine in yeast cells cultured in a medium containing putrescine and sodium dihydrogen phosphate. The amount of polyamine in yeast cells cultured in a medium containing spermidine. The amount of polyamine in yeast cells cultured in a medium containing putrescine and spermidine. The amount of polyamine in yeast cells cultured in a medium containing putrescine and a sodium source. A: Zygosaccharomyces rouxii, B: Candida utilis. The amount of polyamine in yeast cells cultured in a normal medium.

  In the present specification, "polyamine" refers to a linear aliphatic hydrocarbon having a plurality of primary amino groups unless otherwise specified, and the type thereof is not particularly limited. Usually, polyamines mainly produced by yeast are putrescine, spermidine and spermine. Therefore, preferably, "polyamine" in the present specification is a general term for putrescine, spermidine and spermine. Therefore, the term "amount of polyamine contained in yeast" or "amount of polyamine in yeast cells" herein preferably refers to the total amount of putrescine, spermidine and spermine in the yeast cells. The amount of the polyamine in the yeast cells can be measured by an HPLC method (for example, see Reference Examples 1 and 2 described later) and the like.

  The present invention provides a polyamine-rich yeast containing a polyamine at a high concentration in cells. The polyamine-rich yeast of the present invention can be produced by culturing yeast according to the culturing method described below.

  As the yeast to be cultured, yeast used for normal edible or fermentation can be used. Examples of such yeasts include the genus Saccharomyces, Candida, Candida, Torulopsis, Zygosaccharomyces, Schizosaccharomyces (Schizopsiachus), and Schizosciaciasica of the genus Ascomycota. The genus Yarrowia, the genus Hansenula, the genus Kluyveromyces (Kluyveromyces), the genus Debaryomyces, the genus Geotricum (Geotrichum), the genus Wickelhamia, and the genus Wickerhemia, Wikkerhamia Sporobolomyces of the Basidiomycota Yeast belonging to the lomyces), and the like. Among these, yeast belonging to the genera Saccharomyces, Candida and Digosaccharomyces is preferable, and Saccharomyces cerevisiae, Saccharomyces pastorianus, Candida utilis, and Zygosaccharomyces rouxi are more preferable.

In the present invention, a medium to which the following A component or the following B component is added is used as a medium for culturing the yeast.
A component) one or more polyamines selected from the group consisting of putrescine, spermidine and spermine B component) sodium chloride, sodium glutamate, sodium dihydrogen phosphate, citric acid and sodium hydroxide, sodium salt of citric acid, sodium sulfate At least one sodium source selected from the group consisting of: sodium succinate, sodium tartrate, disodium fumarate, and sodium malate

  The component A may be any one kind of polyamine selected from the group consisting of putrescine, spermidine and spermine, a combination of any two or more kinds, or a combination of all kinds. Preferably, putrescine is used. The putrescine, spermidine, and spermine used as the A component may be synthetic or natural products prepared from bacteria, foods, and the like.

  The B component is a sodium source. Examples of the sodium source include sodium chloride, sodium glutamate, sodium dihydrogen phosphate, citric acid and sodium hydroxide, sodium salts of citric acid (eg, monosodium citrate, disodium citrate, trisodium citrate), sulfuric acid One or more selected from the group consisting of sodium, sodium succinate, sodium tartrate, disodium fumarate, and sodium malate can be used. As the sodium source, those sold as reagents can be used.

  In the present invention, the above components A and B may be used alone or in combination. Preferably, yeast is cultured in a medium containing both the A component and the B component. When the A component is used alone, the amount of the A component added to the medium is 0.1 mM or more, preferably 0.3 mM to 30 mM, more preferably, as the total concentration of putrescine, spermidine and spermine in the medium. 0.5 mM to 20 mM. More specifically, the preferable concentration of putrescine in the medium is 0.5 mM or more, more preferably 1 to 50 mM, further preferably 1 to 30 mM, and the preferable concentration of spermidine is 0.1 mM or more, more preferably 0.1 mM. The concentration is preferably 1 to 30 mM, more preferably 0.1 to 20 mM, and the preferred concentration of spermine is 0.1 mM or more, more preferably 1 to 20 mM, and still more preferably 1 to 10 mM. When the B component is used alone, the amount of the B component added to the medium is 100 mM or more, preferably 100 mM to 2 M, more preferably 100 mM to 1 M, as the sodium concentration in the medium. If a medium having a higher sodium concentration is used, the yield of polyamine may decrease.

  When the components A and B are used in combination, the components A and B can be used at the above-described concentrations in the case of single addition. Preferably, when the component A and the component B are used in combination, the concentration of the component A is 0.1 mM or more, more preferably 0.25 mM to 30 mM, and the concentration of the component B is 25 mM or more, preferably 25 mM to 1 M. , More preferably 25 mM to 500 mM. For example, A component is 0.1 mM, B component is 25 mM or more, preferably 25 mM to 1 M, more preferably 25 mM to 500 mM, or A component is 0.25 mM to 30 mM, and B component is 25 mM or more, preferably Is 25 mM to 1 M, more preferably 25 mM to 500 mM.

  When adding the A component to the medium, before or after inoculating the yeast into the medium prepared and autoclaved by the usual procedure, the A component is added to the predetermined final concentration. Good. When the B component is added to the medium, the B component may be added so as to have the predetermined final concentration at the stage of preparing the medium, or the medium prepared in an ordinary procedure and autoclaved, Before or after inoculating the yeast, the B component may be added to the above-mentioned predetermined final concentration.

  In other words, in the present invention, the medium containing the component A and / or B may be inoculated with yeast and cultured, and after inoculating the medium with the yeast, the component A and / or B may be further added. Culture may be carried out with addition, or after inoculating yeast into a medium containing the above-mentioned component A and / or component B, the above-mentioned component A and / or component B may be further added and cultured.

  As the basal medium of the medium containing the component A and / or component B used in the present invention, a medium that can be used for culturing yeast may be used, and any of a synthetic medium, a natural medium, and a combination thereof may be used. It may be. Preferred examples include a YM medium, a YPD medium, an SD medium (minimum glucose medium), a YMPD medium, and a molasses medium (sugar concentration of 0.3 to 10% by mass / volume, for a fermentation test). The typical composition of these media is as follows: YPD medium [10 g / L yeast extract, 20 g / L peptone, 10 g / L glucose]; YM medium [3 g / L yeast extract, 3 g / L malt extract, 3 g / L peptone, 10 g / L glucose]; SD medium [6.7 g / L Yeast Nitrogen base with Amino acid, 20 g / L glucose]; YMPD medium [3 g / L yeast extract, 3 g / L malt extract, 5 g / L] Peptone, 30 g / L glucose]; molasses medium [molasses diluted with water to a reducing sugar concentration of 0.3 to 10% by mass / volume]. Alternatively, an aqueous glucose solution can be used as the basal medium. The glucose concentration of the aqueous glucose solution is preferably 0.5 to 10% by mass / volume. Preferably, the medium is sterilized by autoclaving at 121 ° C. for 15 minutes after preparation. The solidification of the medium may be performed according to a conventional method, such as preparing a plate by adding 2% agar.

  Alternatively, when increasing the polyamine content in the cells while suppressing the growth of yeast or suppressing the growth only slightly, use water or an aqueous solution containing the component A and / or B instead of the above-described medium. Then, the yeast may be cultured.

  The yeast culture medium containing the above-mentioned component A and / or component B is one aspect of the present invention. More specifically, the yeast culture medium is a medium for producing a polyamine-rich yeast.

  In the present invention, a yeast culturing method can be performed according to a usual yeast culturing method except that a medium for producing a polyamine-rich yeast containing the above-mentioned component A and / or component B is used. The yeast culturing method and conditions can be determined with reference to, for example, Microbiol Cult Coll, 25 (2) 89-91, 2009.

  Preferably, the yeast is first inoculated into a normal liquid medium, and precultured while shaking at 30 ° C for 1 to 2 days. If necessary, the precultured medium may be centrifuged or the like to separate the yeast fraction. Next, the above-mentioned component A and / or component B are added to the autoclave-sterilized basal medium to prepare a main culture medium, and then a preculture liquid or a yeast fraction thereof is added thereto, and the mixture is heated at 29 to 32 ° C and 120 to 150 rpm. Culture with shaking (in the case of liquid medium) or static culture at a degree. The amount of the preculture solution or the yeast fraction added to the main culture medium is about 10 to 80% by volume of the preculture solution or 10 to 1500% by volume of the yeast fraction based on 100% by volume of the main culture medium ( It may be about (pre-culture solution equivalent), and can be appropriately changed according to the culture scale, apparatus, environment and the like. The culturing time of the main culture varies depending on the culture scale, but is about 3 hours to 2 weeks. Using the amount of growth of yeast using the degree of suspension of the culture medium as an index and the amount of polyamine in the yeast cells obtained by sampling as an index, determine the culture time that can efficiently produce polyamine on a desired culture scale. be able to.

  According to the present invention, by culturing yeast in a medium to which component A and / or component B has been added as described above, the polyamine productivity of yeast can be improved, and polyamine is contained at a high concentration in cells. A polyamine-rich yeast can be produced. The produced polyamine-rich yeast of the present invention has a higher polyamine productivity and contains a larger amount of polyamine in cells than conventionally known yeasts. Preferably, the polyamine-rich yeast of the present invention contains polyamine in an amount of 150 mg or more, more preferably 200 mg or more, still more preferably 250 mg or more, still more preferably 300 mg or more, and still more preferably 400 mg or more, per 100 g of cells (dry matter). , More preferably in an amount of 500 mg or more.

  The polyamine-rich yeast of the present invention has a high polyamine content in the cells, and is therefore useful as a polyamine source. For example, the polyamine-rich yeast of the present invention can be directly added to foods, nutritional compositions, and the like. Alternatively, a polyamine-containing yeast containing a high concentration of a polyamine is separated from the cells of the polyamine-rich yeast of the present invention by a known method (for example, the method described in Patent Document 4) and, if necessary, purified. A composition can be manufactured. The polyamine-containing composition can be used as an active ingredient in foods, nutritional compositions, pharmaceuticals and the like.

  Hereinafter, the effects of the present invention will be specifically described with reference to Examples and Comparative Examples, but the present invention is not limited to these Examples and the like. The polyamine-rich yeast obtained by an equivalent method with reference to the method described in the present specification is included in the range of the polyamine-rich yeast of the present invention.

  In the following examples, extraction of polyamine from yeast cells and measurement of the amount of polyamine were performed according to the procedures described in Reference Examples 1 and 2, respectively.

(Reference Example 1 Extraction of polyamine from yeast cells)
The medium was centrifuged to collect the bacteria, washed twice with ultrapure water, and then added with ultrapure water to a total volume of 0.5 mL to prepare a bacterial solution. 0.2 mL of the bacterial solution was collected, and the dry weight of the contained bacterial cells was measured. Further, 4.8 mL of 0.2 M perchloric acid was added to 0.2 mL of the bacterial solution, heated at 100 ° C. for 10 minutes, and then centrifuged (4 ° C., 2000 rpm, 3 minutes). The supernatant was filtered with a 0.45 μm filter, and the obtained filtrate was diluted with 0.2 M perchloric acid. 0.5 mL of a saturated sodium carbonate aqueous solution and 1 mL of a 1% dansyl chloride-acetone solution were added to 1 mL of the diluent, mixed, and reacted at 45 ° C. for 1 hour under light shielding. 0.5 mL of 10% proline was added to the reaction solution, mixed well, and allowed to stand still for 10 minutes under light shielding. Then, 2 mL of toluene was added, and the mixture was shaken at 250 rpm for 10 minutes to collect a toluene layer. 1 mL of the toluene layer was dried under reduced pressure at 40 ° C. and dissolved in 1 mL of acetonitrile to prepare a sample for HPLC measurement.

(Reference Example 2 Measurement of Polyamine Amount)
Using a polyamine standard having a known concentration, HPLC measurement was performed under the following conditions, and a calibration curve was prepared from the concentration of each polyamine standard and the area under the peak of the HPLC retention time. Calibration curves were prepared for each of the polyamine standards using putrescine dihydrochloride, spermidine trihydrochloride, and spermine tetrahydrochloride (all from Sigma Aldrich). Next, the sample for HPLC measurement prepared in Reference Example 1 was subjected to HPLC measurement under the same conditions, and peaks of putrescine, spermidine and spermine were detected. From the area under each peak, the amounts of putrescine, spermidine and spermine in the sample were quantified based on the corresponding calibration curve.
(HPLC conditions)
Column: CAPCELL PAK C18 UG-120
3μm, 4.6 × 100mm (Shiseido)
Eluent A: 10 mM ammonium acetate Eluent B: Acetonitrile Gradient program:
Time (minutes) A (%) B (%)
0 50 50
15 10 90
25 10 90
25.1 50 50
35 50 50
Column temperature: 35 ° C
Flow rate: 1.0 mL / min
Detection: Ex. 365 nm, Em. 510 nm

(Example 1)
Baker's yeast (Saccharomyces cerevisiae, Oriental Yeast Co., Ltd.) was inoculated into a test tube containing 5 mL of YMPD medium (3 g / L yeast extract, 3 g / L malt extract, 5 g / L peptone, 30 g / L glucose), Shaking culture was performed at 30 ° C. and 150 rpm for 24 hours. 0.5 mL of the obtained culture was centrifuged (4 ° C., 3000 rpm) to collect the cells. This was inoculated into 5 mL of a YMPD medium supplemented with 10 mM putrescine, and cultured with shaking at 30 ° C. and 150 rpm for 3, 6, 8 or 22 hours. After the cultivation, yeast cells were recovered from the medium and polyamine was extracted according to the procedures described in Reference Examples 1 and 2, and the amount of polyamine in the cells was measured.
FIG. 1 shows the measurement results of the polyamine amount. By culturing in a medium supplemented with putrescine, the amount of polyamine in yeast cells increased.

(Example 2)
The yeast was cultured in the same procedure as in Example 1 except that a YMPD medium supplemented with a sodium source or other salts was used instead of 10 mM putrescine, and the amount of polyamine in the cells was measured. Sodium (Na) sources or salts include 680 mM sodium chloride, sodium glutamate, sodium dihydrogen phosphate, sodium sulfate, phosphate buffer, and sodium bicarbonate (FIG. 2), 100 mM trisodium citrate, and 200 mM Sodium succinate, sodium malate, sodium tartrate, sodium gluconate, disodium fumarate, sodium ascorbate, sodium aspartate, and sodium lactate (FIG. 3), or 350 mM citric acid + sodium hydroxide, lactic acid + Sodium hydroxide and hydrochloric acid + sodium hydroxide (FIG. 4) were used. The above concentrations are all concentrations as sodium. The culture time was 24 hours.
The measurement results of the polyamine amount are shown in FIGS. By culturing in a medium supplemented with a specific type of sodium source, the amount of polyamine in the yeast cells increased.

(Example 3)
Baker's yeast (S. cerevisiae, Oriental Yeast Co., Ltd.) was inoculated into a test tube containing 5 mL of YMPD medium (3 g / L yeast extract, 3 g / L malt extract, 5 g / L peptone, 30 g / L glucose). And shaking culture at 150 rpm for 30 hours at 30 ° C. 2.5 mL of the obtained culture was centrifuged (4 ° C., 3000 rpm) to collect the cells. This was inoculated into 5 mL of YMPD medium supplemented with sodium dihydrogen phosphate (0, 25, 50 or 100 mM) and putrescine (0, 1, 2 or 4 mM), and cultured with shaking at 30 ° C. and 150 rpm for 24 hours. After the cultivation, yeast cells were recovered from the medium and polyamine was extracted according to the procedures described in Reference Examples 1 and 2, and the amount of polyamine in the cells was measured.
FIG. 5 shows the measurement results of the polyamine amount. By adding putrescine and a sodium source in combination, the amount of polyamine in the yeast cells was significantly increased as compared with the case of adding each alone.

(Example 4)
The yeast was cultured in the same procedure as in Example 1 except that a YMPD medium supplemented with trisodium citrate was used instead of putrescine, and the amount of polyamine in the cells was measured. The concentration of trisodium citrate in the medium was 25, 50, 100, and 200 mM. The culture time was 24 hours. FIG. 6 shows the measurement results of the polyamine amount.

(Example 5)
The yeast was cultured in the same procedure as in Example 1 except that a YMPD medium supplemented with trisodium citrate and putrescine was used instead of the addition of putrescine alone, and the amount of polyamine in the cells was measured. The concentration of trisodium citrate in the medium was 0 (only putrescine), 25, 50, 100, and 200 mM, and the concentration of putrescine was 1 mM. The culture time was 24 hours. FIG. 7 shows the measurement results of the polyamine amount. Compared with the case of adding the sodium source alone, the combined use of putrescine and the sodium source significantly increased the amount of polyamine in the yeast cells.

(Example 6)
The yeast was cultured in the same manner as in Example 1 except that a YMPD medium supplemented with sodium dihydrogen phosphate and putrescine was used instead of adding putrescine alone, and the amount of polyamine in the cells was measured. The concentration of sodium dihydrogen phosphate in the medium was 25 or 100 mM, and the concentrations of putrescine were 0.125, 0.25 and 0.5 mM. The culture time was 24 hours. FIG. 8 shows the measurement results of the polyamine amount. The combined use of putrescine and a sodium source significantly increased the amount of polyamine in the yeast cells.

(Example 7)
Yeast was cultured in the same manner as in Example 1 except that a YMPD medium supplemented with spermidine was used instead of putrescine, and the amount of polyamine in the cells was measured. The concentrations of spermidine in the medium were 0.0625, 0.125, 0.5 and 1 mM. The culture time was 24 hours. FIG. 9 shows the measurement results of the polyamine amount.

(Example 8)
Instead of using putrescine alone, a yeast was cultured in the same procedure as in Example 1 except that a YMPD medium supplemented with putrescine and spermidine was used, and the amount of polyamine in the cells was measured. The concentration of putrescine in the medium was 0.25 mM, and the concentrations of spermidine were 0.0625, 0.125, and 0.25 mM. The culture time was 24 hours. FIG. 10 shows the measurement results of the polyamine amount.

(Example 9)
Instead of baker's yeast, Zygosaccharomyces rouxii (NBRC1730) and Candida utilis (NBRC10707) were inoculated, and YMPD medium supplemented with 100 mM sodium dihydrogen phosphate and putrescine 1 mM was used instead of YMPD medium supplemented with 10 mM strepsin. The yeast was cultured in the same procedure as in Example 1 except that the amount of polyamine was measured. The culture time was 24 hours. The measurement results of polyamine are shown in FIGS. 11A and 11B.

(Comparative example)
S. cerevisiae (Oriental Yeast Co., Ltd .; baker's yeast); cerevieiae strains (NBRC2043, NBRC10217); pastrianus (NBRC1961 and NBRC10010) and Cantida utilis (NBRC10707) were inoculated into a test tube containing 5 mL of YMPD medium (3 g / L yeast extract, 3 g / L malt extract, 5 g / L peptone, 30 g / L glucose). And shaking culture at 150 rpm for 30 hours at 30 ° C. 0.5 mL of the obtained culture was centrifuged (4 ° C., 3000 rpm) to collect the cells. This was inoculated into 5 mL of YMPD medium and cultured with shaking at 30 ° C. and 150 rpm for 24 hours. After the cultivation, yeast cells were recovered from the medium and polyamine was extracted according to the procedures described in Reference Examples 1 and 2, and the amount of polyamine in the cells was measured.
FIG. 12 shows the measurement results of the polyamine amount.

Claims (4)

A method for producing a polyamine-rich yeast containing a polyamine in an amount of 150 mg / 100 g or more (in terms of dry matter) , comprising culturing yeast in a medium to which the following A component and / or B component has been added:
A component One or more polyamines selected from the group consisting of putrescine, spermidine and spermine, in a total of 0.1 mM or more;
B component One or more sodium sources selected from the group consisting of sodium chloride, sodium glutamate, sodium dihydrogen phosphate, citric acid and sodium hydroxide, a sodium salt of citric acid, and sodium malate, having a sodium concentration of 25 mM that's all.   The method according to claim 1, wherein both the A component and the B component are added to the medium.   The method according to claim 1 or 2, wherein the yeast is of the genus Saccharomyces. And separating the claims 1 to yeast or we polyamine prepared in any one of claims ways 3, the manufacturing method of the polyamine.