JPH08332083A - Yeast containing iron at high concentration - Google Patents

Abstract

PURPOSE: To obtain the subject yeast useful for perorally administrable pharmaceutical preparations, foods and drinks, feeds, baits, etc., containing iron at a high concentration by culturing a yeast in a suspended state under slowly stirring or shaking in a solution containing a specific amount or more of the iron. CONSTITUTION: This yeast contains iron at a high concentration. The yeast is obtained by using ferrous sulfate, preparing a solution containing >=2000ppm ferrous ions in the solution, then suspending a yeast such as a baker's yeast in the solution, slowly stirring and/or shaking the suspension at 30 deg.C for 24hr, collecting the yeast fungal cell in the suspended state by centrifugation and washing the collected yeast fungal cell with ion exchange water. The resultant yeast contains >=15000ppm iron and bivalent iron excellent in absorbability at a high concentration and is useful as a perorally administrable pharmaceutical preparation or various foods such as breads, biscuits or miso. The cell wall of the resultant yeast is destroyed to take out the contents, which are then used to provide foods and drinks such as a juice, an ice cream or a cracker, feeds or baits containing a large amount of iron.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an edible yeast containing a high concentration of minerals, especially iron, and a method for producing the same.

[0002]

2. Description of the Related Art It is well known that iron is one of the important trace minerals in the living body. However, although dietary habits have become richer and improved significantly in recent years, iron deficiency is widely recognized not only in developing countries but also in developed countries.

In order to improve such iron deficiency, for example, to improve the symptoms of iron deficiency anemia, iron agents such as ferrous sulfate have been conventionally used. However, these iron agents have a drawback in that they are easily oxidized into trivalent iron compounds which are difficult to be absorbed from divalent iron having excellent absorbability during storage or in the digestive tract.

[0004]

SUMMARY OF THE INVENTION In view of the current state of the art, the present invention provides an orally administrable preparation or a food or drink type product containing a high concentration of iron, particularly divalent iron having excellent absorbability. It was made for the purpose of developing a composition.

[0005]

Means for Solving the Problems The present invention has been made in order to achieve the above-mentioned object, and does not directly administer an iron compound as in the prior art, but ingests and accumulates a large amount of iron in edible microorganisms. By using this iron-rich microorganism, we focused on a system that indirectly ingests iron.

[0006] Then, although yeast was cultivated in an iron salt-containing medium by paying attention to yeasts widely used for various edible uses, if the amount of iron salt added was increased, the growth inhibition of yeast was observed according to the conventional technical common sense. However, it was not possible to obtain yeast containing a high concentration of iron.

[0007] However, the inventors of the present invention dared to challenge such common general knowledge and set a new subject of creating a new yeast containing iron at a high concentration which could not be achieved conventionally. Therefore, as a result of examining from various directions, when the yeast was slowly stirred in a suspension state in a high iron concentration solution in a non-proliferating state, the yeast was not cultivated in a nutrient medium. We obtained a new finding that a large amount of iron is taken into the body. Moreover, contrary to the conventional technical common sense, even if the iron concentration is increased, the yeast is not killed, and the iron content in the yeast cells also increases according to the iron concentration in the solution, and 15,000 ppm per dry cell. I confirmed that it would reach the above. Such yeast containing iron at a high concentration has heretofore been considered to be completely uncreatable.

The present invention has been completed as a result of further research based on such useful new knowledge, and the details thereof are as described below.

To carry out the present invention, yeast is suspended in a high-concentration iron-containing solution containing a large amount of a water-soluble salt of iron, and agitated and / or stirred so that the yeast does not settle slowly in a non-proliferative manner.
Or it is necessary to shake. In the present invention,
It does not use a medium for culturing yeast, but treats it non-proliferatively. In other words, yeast is treated under conditions in which substances necessary for the growth of bacterial cells are substantially absent.

As the iron salt used in the present invention, as long as it has appropriate water solubility and does not kill yeast, it can be efficiently taken up in yeast cells.
One or more of all iron compounds, organic or inorganic, can be used. Non-limiting examples of which include: ferrous sulfate, ferrous chloride, ferrous citrate,
Ferrous fumarate, ferrous gluconate, ferrous butyrate, ferrous tartrate, ferrous lactate.

The addition amount of the iron salt is higher than 1,500 ppm, preferably 2,000 ppm or more. If yeast is treated with iron at a concentration as high as 2,000 ppm, the yeast may be killed, or even if it is not killed, serious adverse effects may be suspected. Not surprisingly, it was the first time that yeast was not killed by such a high concentration of iron, but it was the first time that it could be taken up into the cells. Moreover, it has been confirmed that treatment with iron having a high concentration of 10,000 ppm is possible.

As a result, 15,000 p per dry cell
Finally, we succeeded in incorporating yeast of pm or more into yeast cells. Yeast containing 10,000 ppm or more of iron has not existed in the past, and is a novel useful yeast that could not be created even if it was attempted to be created as described above.
Even more surprisingly, according to the present invention, 10,000 p
30,000 pp by using pm iron solution
It is even possible to incorporate an extremely large amount of iron of m or more into yeast cells.

The treatment conditions for incorporating iron into the yeast cells are as follows.
C., preferably 20 to 35.degree. C., 0.5 to 72 hours, preferably 1 to 36 hours, pH 4 to 7, preferably pH 4.5 to 6.5, but the type of yeast and iron salt used. It may be appropriately determined depending on the concentration, and may be out of the above range in some cases.

The stirring and / or shaking must be carried out at a slow speed so that the yeast and iron ions come into contact with each other,
500 rpm, depending on the viscosity of the iron solution and the yeast content
Below, preferably a gentle agitation of 200 rpm or less,
Or, when the amplitude is 12 cm, it is preferable to shake at 200 rpm or less, preferably about 150 rpm or less.

In the present invention, any yeast can be used as long as it is a yeast, including edible yeast. Non-limiting examples include yeasts belonging to each of the following genera: Saccharomyces, Torrlopsis (Tor).
ulopsis genus, Mycotorula genus, Torulaspora genus, Candida genus,
Rhodotorula genus, Pichia genus and others.

Specifically, Saccharomyces cerevisiae,
Uvarum, rouxii IFO 0439; Torulopsis utilis, same
candida IFO 0856; Mycotorula japonica, same lipolytic
a; Torulaspora delbrueckii, fermentati; Candida
sake, tropicalis, utilis ATCC 16321; Hansenula
anomala IFO 0140, suaveolens; Saccharomycopsis
fibligera IFO 1665, same lipolytica ATCC 20182; Rhodo
torula rubra IFO 0870; Pichia farinosa IFO 0607,
Other yeasts are exemplified.

Among these yeasts, especially as edible yeasts, Saccharomyces cerevisiae and uvaru are commonly used as yeasts for human food and drink, animal feed and fish feed.
Examples include baker's yeast, brewer's yeast, wine yeast, sake yeast, alcohol yeast, amazake yeast, miso soy sauce yeast, feed yeast, and feed yeast, which belong to m, rouxii, Saccharomycopsis fibligera and others.

Water is usually used as the liquid in which the yeast is suspended. The water may be distilled water or deionized water, but tap water is sufficient. The yeast cell concentration is, for example, 10 to 40%, preferably 10 to 20% as the compressed yeast, but is appropriately selected so as to efficiently take in iron. Then, the cells are slowly stirred and / or shaken to the extent that iron and / or yeast cells do not precipitate.

The yeast thus loaded with iron is collected by centrifugation, filtration, etc., and then washed to obtain the desired bacterial cells, and if necessary, freeze-dried or dried cells according to a conventional method. , Used for various purposes. Further, if desired, it is also possible to crush the cells or lyse the cells to take out the contents and use them.

Further, in the present invention, yeast containing a large amount of various minerals in advance is aerated in a solution containing 2,000 ppm or more of iron, as in the case of the above-mentioned ordinary yeast. By slowly stirring and / or shaking in a suspended state without using any of the above, it is possible to create a novel yeast containing a large amount of multi-minerals that further contains iron in a high concentration in various minerals.

Examples of yeasts with a high content of multi-minerals include yeasts with a high content of zinc and iron, yeasts with a high content of magnesium and iron, yeasts with a high content of magnesium, zinc and iron, and the like.

The zinc-rich iron-rich yeast is produced by using the previously-produced zinc-rich yeast as a raw material yeast, and incorporating iron into the yeast under the above-mentioned iron-rich yeast production conditions according to the present invention. You can

Yeasts high in zinc are produced under the conditions for producing yeasts high in zinc, that is, in a solution containing 50 ppm or more of zinc.
It can be produced by non-proliferatively stirring and / or shaking yeast in suspension.

In order to carry out the present method, it is necessary to suspend the yeast in a high-concentration zinc-containing liquid containing a large amount of a water-soluble salt of zinc, and agitate and / or shake it non-proliferatively. . In this method, a medium for culturing yeast is not used, but a non-proliferative treatment is performed. In other words, yeast is treated under conditions in which substances necessary for the growth of bacterial cells are substantially absent.

As the zinc salt to be used, any zinc salt, organic or inorganic, can be used as long as it has appropriate water solubility, does not kill yeast, and can be efficiently taken up in yeast cells. One or more compounds can be used. Non-limiting examples include: zinc sulfate, zinc chloride, zinc citrate, zinc acetate, zinc tartrate, zinc lactate.

A good result can be obtained even when the amount of the zinc salt added is 50 ppm, but a higher concentration than that, 2,000 ppm or more, particularly preferably 3,000 to 6,000 ppm is suitable. According to the conventional technical common sense, if yeast is treated with a high concentration of zinc, the yeast may be killed, or even if it is not killed, there is a serious fear of serious adverse effects. The present invention is significantly transcendental, and, unexpectedly, it was the first time that yeast was not killed by such a high concentration of zinc, but rather it was first succeeded in incorporating it into the cells. is there. Furthermore, it has been confirmed that treatment with zinc having a high concentration of 10,000 ppm or more is possible.

As a result, 600 to 10 per dry cell,
It was possible for the first time to incorporate zinc of 000 ppm or more into yeast cells. Even more surprisingly, according to the present invention, by using a 10,000 ppm zinc solution, it is even possible to incorporate a very large amount of zinc of 30,000 ppm or more per dry cell into yeast cells. .

The treatment conditions for incorporating zinc into the yeast cells are as follows.
The temperature is 0 ° C., preferably 20 to 35 ° C., 1 to 120 hours, preferably 40 to 100 hours, pH 4 to 7, preferably pH 4.5 to 6.5. It may be appropriately determined depending on the concentration, and may be out of the above range in some cases.

The stirring and / or shaking depends on the viscosity of the zinc solution, the yeast content, etc., but is 500 to 1,000 rpm.
If the stirring is moderate, or the amplitude is 12 cm, 100-
It is recommended to shake at 200 rpm. Good results can be obtained by performing aeration under aerobic conditions. Aeration is performed at about 0.5 to 5 vvm, but if necessary, the range may be deviated.

As described above, a zinc-iron-rich yeast can be created. As a result, a novel multi-mineral edible yeast containing 10,000 ppm of zinc and 15,000 ppm or more of iron per dry cell can be obtained.

The magnesium-rich iron-rich yeast is produced by using the previously-produced magnesium-rich yeast as a raw material yeast and treating iron with yeast under the above-mentioned iron-rich yeast production conditions according to the present invention. You can

The high magnesium content yeast is produced by non-proliferative stirring and / or shaking of the yeast in suspension in a high magnesium content production condition, that is, in a solution containing a high concentration of magnesium. be able to.

In order to carry out the present method, it is necessary to suspend the yeast in a high concentration magnesium-containing solution containing a large amount of a water-soluble salt of magnesium, and aerobically agitate and / or shake it. . In this method, the magnesium concentration in the finally created yeast cells depends on the magnesium concentration in the solution in which the yeast is suspended, regardless of the composition of the medium for culturing the yeast. In other words, it is possible to allow magnesium to be incorporated into yeast even under the condition that a substance necessary for the growth of bacterial cells exists or under the condition that it does not substantially exist.

The magnesium salt used in the present method may be organic or inorganic, as long as it has appropriate water solubility, does not kill yeast, and is efficiently taken up by yeast cells. All magnesium compounds can be used alone or in combination. Non-limiting examples of which include: magnesium sulfate, magnesium chloride, magnesium citrate, magnesium acetate, primary magnesium tartrate, primary magnesium lactate.

The amount of magnesium salt added is preferably set so that the magnesium concentration in the liquid is 50 ppm or more. If yeast is treated with such a high concentration of magnesium, the yeast may be killed, or even if it is not killed, serious adverse effects such as reduced activity may be suspected. The present invention is significantly transcendental and, unexpectedly, not only yeast is not killed by such high concentration of magnesium, but extremely high concentration of magnesium can be accumulated in the bacterial cells, and We obtained a useful new finding that the physiological activity of P.

As a result, 2,000 pp per dry cell
Finally, we succeeded in incorporating more than m of magnesium into the yeast cells. Even more surprisingly, according to this method, by using a solution containing 20,000 ppm of magnesium ions, it is possible to obtain 20,000 per dry cell.
It is even possible to incorporate a very large amount of magnesium of 0 ppm or more into yeast cells.

In order to make a large amount of magnesium contained in yeast cells according to this method, 5 to 30%, preferably 10 to 20% of yeast as wet cells are suspended in a magnesium solution and the temperature is set to 15 to 40 ° C. , Preferably 25-35
C., more preferably 28 to 33.degree. C., treatment time 1 to 120 hours, preferably 36 to 96 hours, pH 4 to 4.
7, preferably pH 4.5-6.0, aeration 0-5vv
m, preferably 1 to 3 vvm, stirring varies depending on the scale of equipment used, but 50 to 1,000 rpm, and 100 to 200 rp for equipment having an amplitude of 12 cm when shaking.
m, preferably 100 to 150 rpm, but can be appropriately selected depending on the type of yeast and magnesium salt used and the concentration thereof, and may deviate from the above range depending on the necessity.

[0038] As described above, the magnesium-iron-rich yeast can be produced. As a result, magnesium per dry cell was 2,000 ppm and iron was 15,000.
A novel multi-mineral edible yeast containing 0 ppm or more can be obtained.

The magnesium / zinc / iron-rich yeast is prepared by using a previously-produced magnesium / zinc-rich yeast as a raw material yeast and incorporating iron into the yeast under the above-described iron-rich yeast production conditions according to the present invention. Can be manufactured by.

The magnesium / zinc-rich yeast is produced by using the magnesium / zinc-rich yeast production conditions, that is, the magnesium-rich yeast obtained under the above-described magnesium-rich zinc production conditions as a raw material yeast, in a high zinc concentration. It can be produced by stirring and / or shaking yeast in a suspension containing it in a suspension state non-proliferatively.

In this way, a yeast containing a high content of magnesium, zinc and iron can be created. As a result, magnesium per dry cell is 2,000 ppm and zinc is 1
It is possible to obtain a novel multi-mineral edible yeast containing 10,000 ppm and iron of 15,000 ppm or more.

Examples of the present invention will be described below.

[0043]

Example 1 Using ferrous sulfate (FeSO ₄ ), a solution was prepared so that divalent iron ions would be 100 to 10,000 ppm in the solution (Table 1 below), and an iron solution of each concentration was prepared. 1
20 g (2 × 10 ⁹ cells / ml) of baker's yeast (Regular yeast manufactured by Oriental Yeast Co., Ltd.) was suspended in 00 ml, and shaken in a 500 ml Sakaguchi flask at 30 ° C. for 24 hours (amplitude 12 cm, 110 rpm). .

Centrifugation of the cells after shaking (3,000 x
g, 5 minutes) to collect the bacteria, and deionized water (Milli)
After washing twice with Q), the cells were wet-ashed and iron was quantified by an atomic absorption spectrophotometer. In addition, the amount of residual iron in the combined solution of the supernatant after washing the cells and the washing solution was colorimetrically determined by the orthophenanthrin method. The obtained results are shown in Table 1 below.

[0045]

[Table 1]

[0046]

Example 2 A solution was prepared using ferrous sulfate (FeSO ₄ ) so that the divalent iron ion was 5,000 ppm in the solution, and 1.5 L of this iron solution was added to baker's yeast (Oriental yeast). Industrial Co., Ltd. regular yeast) 300g
(2 × 10 ⁹ cells / ml) was suspended and gently stirred in a 3 L jar fermenter at 30 ° C. for 24 hours at 200 rpm at which yeast did not precipitate.

After stirring for a predetermined time, the bacterial cells were collected by centrifugation (3,000 xg, 5 minutes), washed twice with ion-exchanged water (MilliQ), wet-ashed and then atomized. Iron was quantified by an absorption spectrophotometer. In addition, the amount of residual iron in the combined solution of the supernatant after washing the cells and the washing solution was colorimetrically determined by the orthophenanthrin method. The obtained results are shown in FIG.

[0048]

Example 3 Using zinc sulfate (ZnSO ₄ .7H ₂ O),
So that the zinc ion is 5,000 ppm in the solution,
A solution was prepared, and baker's yeast (Regular yeast manufactured by Oriental Yeast Co., Ltd.) 300
g (2 × 10 ⁹ cells / ml) was suspended, and the mixture was stirred in a 3 L jar fermenter with aeration of 2.0 vvm and 650 rpm, and treated at 30 ° C. for 72 hours under non-growth conditions.

After aeration and stirring for a predetermined time, the cells were collected by centrifugation (3,000 xg, 5 minutes) and washed twice with ion-exchanged water (MilliQ), and then the cells were wet ashed. As a result of quantifying zinc with an atomic absorption spectrophotometer,
It contained 14,500 ppm of zinc per dry cell.

Using the zinc-rich yeast thus obtained as a starting yeast, ferrous sulfate (FeSO ₄ ) was used to prepare a solution so that the divalent iron ion would be 5,000 ppm in the solution. 300 L of raw material yeast (2 x 1) containing a high concentration of zinc prepared in advance in 1.5 L of this solution.
( ⁹ cells / ml) and suspended in a 3 L jar fermenter at 30 ° C. for 24 hours for 20 hours at which yeast does not precipitate.
The mixture was gently stirred at 0 rpm.

After stirring for a predetermined time, the bacterial cells were collected by centrifugation (3,000 × g, 5 minutes), washed twice with ion-exchanged water (MilliQ), wet-ashed and then atomized. Iron and zinc were quantified by an absorption spectrophotometer.
The obtained results are shown in Table 2.

[0052]

[Table 2]

[0053]

Example 4 Magnesium sulfate (MgSO ₄ · 7H ₂ O)
With magnesium ion in solution at 15,000p
A solution was prepared so as to have pm, and 300 g (2 × 10 ⁹ cells / ml) of baker's yeast (Regular yeast manufactured by Oriental Yeast Co., Ltd.) was added to 1.5 L of this magnesium solution.
And aeration with a 3 L jar fermenter 2.
The mixture was stirred at 0 vvm and 650 rpm and treated at 30 ° C. for 72 hours.

After aeration and stirring for a predetermined time, the bacterial cells were collected by centrifugation (3,000 xg, 5 minutes), washed twice with ion-exchanged water (MilliQ), and then wet-ashed. As a result of quantifying magnesium with an atomic absorption spectrophotometer, it was found that 7,830 ppm of magnesium was contained per dry cell.

Using the magnesium-rich yeast thus obtained as a raw material yeast, ferrous sulfate (FeSO ₄ ) was used to prepare a solution so that divalent iron ions would be 5,000 ppm in the solution. Then, 300 g (2 × 10 ⁹ pieces / ml) of the raw material yeast containing a high concentration of magnesium prepared in advance was suspended in 1.5 L of this iron solution, and the yeast was suspended in a 3 L jar fermenter at 30 ° C. for 24 hours. The mixture was gently stirred at 200 rpm at which it did not settle.

After stirring for a predetermined time, the cells were collected by centrifugation (3,000 xg, 5 minutes), washed twice with ion-exchanged water (MilliQ), wet-ashed, and then atomized. Iron and magnesium were quantified by an absorption spectrophotometer. Table 3 shows the obtained results.

[0057]

[Table 3]

[0058]

Example 5 Using zinc sulfate (ZnSO ₄ .7H ₂ O),
So that the zinc ion is 5,000 ppm in the solution,
A solution was prepared, and 300 g (2 × 10 ⁹ cells / ml) of the magnesium-rich yeast produced according to the conditions described in Example 4 was suspended in 1.5 L of this zinc solution, and the suspension was placed in a 3 L jar fermenter. Aeration 2.0 vvm, 650r
The mixture was stirred at pm and treated at 30 ° C. for 72 hours under non-growth conditions.

After aeration and stirring for a predetermined time, the cells were collected by centrifugation (3,000 xg, 5 minutes), washed twice with ion-exchanged water (MilliQ), and then wet-ashed. Quantitative analysis of magnesium and zinc with an atomic absorption spectrophotometer showed that magnesium per dry cell was 6,57
Yeast cells simultaneously containing 0 ppm and 11,300 ppm of zinc were obtained.

The magnesium / zinc-rich yeast thus obtained was used as a starting yeast, and ferrous sulfate (FeSO 4
₄ ), divalent iron ions in the solution are 5,000pp
A solution was prepared so that the amount of m becomes m, and 300 g (2 × 10 ⁹ pieces / ml) of the raw material yeast containing a high concentration of magnesium and zinc prepared in advance was suspended in 1.5 L of this iron solution, and 3 L jar fur was prepared. With a mentor, the mixture was gently stirred at 30 ° C. for 24 hours at 200 rpm at which yeast did not settle.

After stirring for a predetermined time, the bacterial cells were collected by centrifugation (3,000 × g, 5 minutes), washed twice with ion-exchanged water (MilliQ), wet-ashed and then atomized. Magnesium, zinc and iron were quantified by an absorption spectrophotometer. The results obtained are shown in Table 4.

[0062]

[Table 4]

[0063]

Example 6 Under the same conditions as described in Example 2,
After the yeast treated for 24 hours was collected and washed, a high iron content yeast was obtained. Using this iron-rich yeast, a bread-making test was conducted with the bread composition shown in Table 5 below. As a control, baker's yeast (regular yeast manufactured by Oriental Yeast Co., Ltd.), which was used as a raw material yeast for the iron-rich yeast, was used. Also,
The resulting bread was wet ashed and iron was quantified by an atomic absorption spectrophotometer. The obtained results are shown in Table 6.

[0064]

[Table 5]

[0065]

[Table 6]

[0066]

According to the present invention, 15
It became possible for the first time to make yeast contain a large amount of iron of 000 ppm or more. Such a yeast containing a large amount of iron cannot be obtained in the past and is a completely novel yeast.

The iron-rich yeast created by the present invention can be widely used for various purposes, and various foods such as bread, biscuits, and miso containing a large amount of iron can be obtained. In addition, the cell wall of yeast can be destroyed, the contents can be taken out, and it can be used for the production of various foods and drinks such as juice, ice cream, crackers, and also for the production of feed and feed containing a large amount of iron. can do.

Further, in the present invention, the conventional system of culturing yeast is not adopted, but a completely new system of stirring yeast in a high concentration aqueous solution of iron salt is adopted. Therefore, not only does the need for a medium not only reduce costs, but because it is an aqueous solution of iron salts, washing the yeast cells after harvesting is extremely easy, and separation of the cells can be achieved. Purification can also be carried out very easily.

Furthermore, according to the present invention, various minerals in addition to iron can be contained in yeast in a large amount. Therefore, in addition to iron, minerals such as zinc and / or magnesium can be highly concentrated. It is also possible to create new yeast containing yeast (yeast with high content of multi-minerals), which can be used for various purposes.

[Brief description of drawings]

FIG. 1 shows the relationship between iron concentration in dried yeast cells and treatment time.

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[Procedure amendment]

[Submission date] November 1, 1995

[Procedure Amendment 1]

[Document name to be amended] Statement

[Name of item to be corrected] Brief description of the drawing

[Correction method] Change

[Correction content]

[Brief description of drawings]

FIG. 1 shows the relationship between iron concentration in dried yeast cells and treatment time.

Claims (5)

[Claims] 1. An edible yeast containing a high concentration of iron. 2. Iron is 15,000 ppm per dry cell.
An edible yeast containing the above. 3. Iron in a solution containing 2,000 ppm or more of iron, wherein the yeast is slowly stirred and / or shaken in a suspended state, and the amount of iron per dry cell is 15.0.
A method for producing edible yeast containing at least 00 ppm. 4. The method according to claim 3, wherein yeast containing a high concentration of minerals is used as yeast. 5. The method according to claim 4, characterized in that the mineral is zinc and / or magnesium.