JPH06237778A - Preparation of phytin - Google Patents

Abstract

PURPOSE: To provide a new process for producing phytin comprising the steps of inoculating a yeast into a culture medium of a steep water of corn (LSW) or its concentrated liquid (CSL), culturing the yeast therein to precipitate phytin and collecting it.

CONSTITUTION: A fungus belonging to a lower class which is a unicellular fungous having fermentation activity, used mainly for preparing wine or bread and having no chlorophyll, and which is circular or oval in its shape and multiplies by yeast budding is inoculated into a culture medium of LSW or CSL (in the case of LSW, the concentration is 3-13 BX, and pH is 3-4) at a concentration of 1-20% (V/V) based on the volume of the culture medium), and the fungus is cultured. When the final pH of fermentation beer is about 6.0-8.5 in the case of LSW or about 5-8 in the case of CSL, optionally a magnesium salt or a calcium salt is added in an amount of 0.01-2.0% or 0.01-2.0%, respectively, and the fermentation is continued to precipitate phytin. The speed of precipitation is larger and the yield is better than those in a conventional case.

COPYRIGHT: (C)1994,JPO

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing phytin, more specifically, corn dipping waste liquid (Light St
eep Water, hereinafter referred to as "LSW") or its concentrate (C
Orn Steep Liquor (hereinafter referred to as “CSL”) is used as a medium for inoculating and culturing yeast to produce phytin. Here, yeast is a single cell without chlorophyll that is mainly used for fermenting to make sake or bread, and means a lower fungus that has a circular or elliptical outer shape and grows by yeast budding.

[0002]

2. Description of the Related Art Generally, phytin is a phosphorus storage material in plants and is known to play an important role in germination and early growth of seeds. Phytin is also known as Inositol hexakisphos
phate) calcium, magnesium, zinc, sodium salts, which means mainly calcium and magnesium salts.
During the germination process of seeds, phytin is decomposed by phytase and used for phosphorylation of nucleic acid or membrane components. Phytin is an odorless white powder that is well soluble in weak acids (pH 2-3) and insoluble in water.

A method for producing water-soluble phytin by changing the composition of metal ions is disclosed in Japanese Patent Publication No. 43-1697.
It is published in No. 8. Such phytin is a raw material for producing inositol by hydrolysis at high temperature and high pressure, and phytic acid from which metal ions have been removed is a light yellow or light brown syrup liquid, which is well dissolved in water, 95% ethanol or acetone, Medicinal, fermentation composition, blackening prevention in canning, antioxidant action, water softener, metal corrosion prevention,
It is used in the formation of metal coatings, dye additives, concentration of rare earth elements, polymer solvents, explosion-proof agents such as gasoline, kerosene or fuel oil.

The conventional method for producing phytin is as follows.
In most cases, a precipitate is formed by adding a base such as calcium hydroxide or caustic soda to SW or rice bran to neutralize the final pH to about 7.0-8.0. (Japanese Patent Publication No. 46-35075, No. 46-2
No. 6346, No. 46-33958, No. 42-1726
No. 8 and US Pat. No. 4,163,010), phytic acid used a method in which rice bran or the like was treated with an acid to extract phytic acid, and the phytic acid was passed through an ion exchange resin tower for separation ( U.S. Pat. No. 4,668,813, Japanese Unexamined Patent Publication No. 61-056142, and People's Republic of China Published Patent No. 8602238). Also, Japanese Patent Laid-Open No. 61-0
No. 50989 describes a method for purifying phytic acid by adding a process using an ultrafiltration membrane.

[0005]

However, when phytin is produced by the conventional neutralization method, the formed precipitate is difficult to settle, the viscosity becomes high after neutralization, and it is difficult to separate and dry the precipitate. In addition, there is a problem that when the ion exchange resin tower is used, the process operation is not easy and the production cost is high. Hereinafter, the precipitate means phytin. As a result of extensive researches conducted by the present inventors to solve these problems, the LSW
The present invention has been completed by utilizing the principle that phytin precipitates when the pH increases during yeast culture. Therefore, it is an object of the present invention to provide a novel method for producing phytin.

[0006]

In order to achieve such an object, the method of the present invention comprises inoculating a CSL or LSW medium with yeast and fermenting it to precipitate phytin.

The structure of the present invention will be described in more detail below. The medium used in the present invention is LSW or CSL medium, and LSW is 3 to 13 Bx. When the one having an initial pH of 3 to 4 is used at the above concentration, the yeast grows most vigorously under the above conditions. CSL is 40 to 55 Bx. Of 3 to 20 Bx. It is preferable to use one having a concentration of.

In the present invention, the liquid medium for culturing the yeast to be inoculated is YPD medium or 5-10 Bx.
YSW medium is used for yeast extract (Ye
astExtract) 1% (w / v), glucose 5% (w / v
v) and peptone 1% (w / v). The yeast cultivated in the liquid medium for about 1 day was inoculated into the medium according to the present invention at a concentration of 1 to 20% (v / v) of the volume of the medium, and
Incubate at -40 ° C for about 1 to 4 days. Inoculation amount is 20%
If it exceeds (v / v), the medium is diluted and the amount of precipitation becomes too small. Culture is 100-50
Culture with shaking at about 0 rpm. The final pH after culturing is LS
It is about 6.0 to 8.5 in the case of W and about 5 to 8 in the case of the CSL diluted medium.

As the yeast, any yeast can be used as long as it can grow in the medium using lactic acid as a carbon source. Specifically, Saccharomyces sp. , Candida s
p. , Saccharomycopsis sp. , Schwanniomyces sp. , Ster
igmatomyces sp. , Sympodiomyces sp. , Zygosaccharomy
ces sp. , Saccharomycodes sp. , Gteromyces sp. , Nema
tospora sp. , Sporobolomyces sp. , Kluyveromyces s
P. , Arxiozyma sp. , Lodderomyces sp., etc. are preferable, but not limited thereto. When the pH after culturing becomes 6.0 or more, precipitation of phytin starts, and this precipitation easily precipitates and can be easily separated from the culture supernatant. The sterility of the medium does not significantly affect precipitation formation and yeast growth.

In the present invention, in the case of CSL dilution medium,
Although the amount of the precipitate produced by culturing yeast is very small, calcium salt such as calcium chloride, calcium carbonate or calcium hydroxide, or magnesium salt such as magnesium carbonate hydroxide or magnesium chloride is added in an amount of 0.01 to The addition of 2.0% (w / v) concentration can significantly increase the precipitation formation. If it is less than 0.01%, a sufficient addition effect does not appear, while if it exceeds 2.0%, the pH of the medium becomes too high and the same problem as in the conventional neutralization method occurs. The amount of phytin produced when the calcium salt and the magnesium salt are added is remarkably increased, and the sedimentation property is excellent and the separation is easy. In the case of LSW medium, the addition of calcium salt does not significantly affect the yield of the precipitate, and the precipitate obtained by adding calcium is slightly brown.

In order to compare the sedimentation rate of the precipitate according to the present invention with that of the conventional method, the phytin produced by fermentation and the phytin produced by neutralization were put into a 50 ml graduated cylinder together with the solution, and the sedimentation rate was measured. As shown in 1, the phytin produced by fermentation was almost completely precipitated within 10 minutes, whereas the phytin produced by neutralization was not precipitated at all. Further, when phytin is separated by the fermentation method, the yeast consumes the lactic acid contained in LSW or CSL, and therefore, when the LSW and CSL are concentrated, the viscosity increase is greatly alleviated, which facilitates the concentration step, The degree of enrichment can be greatly increased. Furthermore, when the concentrated liquid is used as a feed, the addition of yeast enhances the nutritional value and eliminates the sourness, so that the effect of improving the quality of the feed can be obtained.

The method for producing phytin by the fermentation method according to the present invention is not limited to the batch culture method such as the flask culture, and the raw material medium 1 is continuously charged into the first fermentor 2 as shown in FIG. Second fermentation tank 3 with partially fermented fermentation products
In the semi-continuous fermentation method in which the remaining fermentation is carried out in a batch system, and as shown in FIG. 3, the raw material medium 1 is put into the fermentation tank 4 and the fermentation product is discharged from the fermentation tank 4 continuously. It can also be carried out by a continuous fermentation method.

According to the present invention, the yield of the precipitate obtained after fermentation is about 1 to 2.5% (w / v) in the case of LSW and 1 to 3% (in the case of CSL diluted medium). w / v). This amount is 85% of the precipitate produced by neutralization.
~ 100% hit. After fermentation is completed, the fermentation liquor is transferred to a separatory funnel and left for about 10 minutes to separate the precipitate.
It can be carried out by separating the precipitated precipitate, washing with distilled water and then drying. The precipitate thus obtained was dissolved in an acid and subjected to Phytate analysis. As a result, the purity was 60 to 95%, which was higher than that of the precipitate obtained by neutralization. In particular, the precipitate obtained by adding calcium carbonate to a CSL-diluted medium and culturing it has a purity of 90%.
It was much higher than the precipitate obtained by the neutralization method (purity ≈ 70%). The above analysis was performed by the method of Ali I. Mohamed (Cereal Chem. 63 (6), 475-8,
1986).

[0014]

EXAMPLES Hereinafter, the effects of the present invention will be described more specifically with reference to Examples, but the scope of the present invention is not limited to the following examples.

Example 1: Concentration 9.2 Bx. LSW10
0 ml was placed in a baffle flask, and 5 ml of a pre-prepared inoculum ( Saccharomyces uvarum NRRL Y-236) was added,
The culture was performed with shaking at 30 ° C and 150 to 350 rpm. Final p
When H became 7.5 or more, the fermentation broth was poured into a separating funnel tube and allowed to stand for 10 minutes to separate the precipitated precipitate. After adding 100 ml of distilled water to the precipitate and shaking it well, it was poured into a separating funnel again to cause sedimentation, and the precipitate was separated, dried in a drying oven at 100 ° C., and weighed to obtain 1.03 g of the precipitate. I got a thing. The purity of the precipitate was 85%, and the metal ion ratio of the precipitate was as shown in Table 1 below.

Example 2: Concentration 9.2 Bx. LSW10
Place 0 ml in a 500 ml flask and add 0.5 g Ca.
After adding CO ₃ (anhydrous), inoculum ( Sa
5 ml of ccharomyces cerevisiae LA) is added and cultured,
When the final pH reached 7.5 or more, the precipitate was removed, and finally 1.30 g of a 79% pure precipitate was obtained. The metal ion ratio of the precipitate is shown in Table 1 below.

Example 3: Concentration 52Bx. CSL of 12Bx. And 0.5 g of C
After dissolving aCl ₂ · 2H ₂ O, inoculum ( Kl
5 ml of uyveromyces lactis ATCC 56498) was added and cultured. After culturing for 2 days, when the pH became 4.9, the precipitate was removed to obtain a precipitate having a purity of 75%. The metal ion ratio of the precipitate is shown in Table 1 below.

Example 4: In Example 3, CaCl ₂
-In place of 2H ₂ O and Kluyveromyces lactis ATCC 56498, CaCO ₃ and Saccharomyces cerevisiae TJ-1 were added,
After 2 days, when the final pH reached 6.40, it was removed to obtain 1.04 g of a 95% pure precipitate. The metal ion ratio of the precipitate is shown in Table 1 below.

Example 5: Concentration of 9.2 Bx. Inoculum of inoculum ( Candida lusitaniae ATCC 34449) using 1 liter of LSW as a raw material, the LSW solution was continuously supplied so that the residence time was 15 hours, and the intermediate fermentation liquid overflowing from the first fermentation tank was subjected to the second fermentation. The solution was received in a tank and cultivated in a batch system until the pH reached 7.5. After the culture was completed, the culture solution in the second fermenter was removed to separate the precipitate. By such a semi-continuous fermentation method, 12 g of precipitate was obtained per liter of raw material LSW,
Its purity was 82%. The metal ion ratio of the precipitate is shown in Table 1 below.

Example 6: 1 in a reactor having a total volume of 3 liters
Lactic acid ( Saccharomyces diastaticus ATCC 13007) was inoculated and cultured with 1 liter of LSW (9.2Bx.) As a raw material, and when the pH reached 7.0, 0.1 liter of the raw material was supplied per hour and the fermentation liquid was added. Fermentation method for extracting 0.1 liters each (dilution ratio = 0.1 hours)
As a result, 8-9 g of a precipitate was obtained per liter of the raw material LSW, and the purity of the precipitate was 80%. The metal ion ratio of the precipitate is shown in Table 1 below.

Comparative Example 1: Concentration 9.2 Bx. LSW10
0 ml was titrated with 5% (w / v) calcium hydroxide,
It was filtered when the final pH was 7.0. After filtration, it was washed with distilled water and dried in an oven at 105 ° C for 3 hours.
5 g of precipitate was obtained. The metal ion ratio of the precipitate is shown in Table 1 below.

Comparative Example 2: Concentration 52Bx. CSL 50m
3.5 g of a precipitate was obtained from 1 by the same method as in Comparative Example 1. The metal ion ratio of the precipitate is shown in Table 1 below.

[0023]

[Table 1]

As a result of analyzing the metal ions of the precipitates of Examples and Comparative Examples, as shown in Table 1, the metal ions of phytin produced by the neutralization method were mainly composed of calcium ions. In contrast, phytin produced by the fermentation method was found to have magnesium ions as the main species, and when fermented by adding calcium carbonate at a concentration of 0.5%, the precipitate formed was Had a significant increase in calcium ion concentration. The metal ion content in Table 1 above was measured by an atomic absorption photometer (Atom) after dissolving the precipitate in an acid.
ic Absorption Spectroscopy). Further, although yeast was mainly used in the above-mentioned examples, all inoculum capable of using lactic acid as a carbon source exhibits a precipitation effect, and the yeast used in the above-mentioned examples does not limit the scope of the present invention.

[0025]

Industrial Applicability As described above, the precipitate produced by the method of the present invention has a very excellent sedimentation property, is easy to separate, has a high phytin purity, and has a high viscosity. Resolved

[Brief description of drawings]

FIG. 1 is a graph comparing the sedimentation rates of phytin according to the present invention and the prior art.

FIG. 2 is a process diagram schematically showing a semi-continuous fermentation method according to the present invention.

FIG. 3 is a process diagram schematically showing a continuous fermentation method according to the present invention.

[Explanation of symbols]

 1 Raw Material Medium 2 First Fermenter 3 Second Fermenter 4 Fermenter

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Claims (12)

[Claims] 1. A method for producing phytin, which comprises inoculating yeast into a CSL or LSW medium and fermenting it to precipitate phytin. 2. The LSW concentration is 3 to 13 Bx. The method for producing phytin according to claim 1, wherein 3. The concentration of CSL is 3 to 20 Bx. The method for producing phytin according to claim 1, wherein 4. The method for producing phytin according to claim 1, wherein 0.01 to 2.0% of magnesium salt is further added to the CSL medium for fermentation. 5. The method for producing phytin according to claim 4, wherein the magnesium salt is magnesium carbonate hydroxide or magnesium chloride. 6. The method for producing phytin according to claim 1, wherein 0.01 to 2.0% of a calcium salt is further added to the CSL medium for fermentation. 7. The method for producing phytin according to claim 6, wherein the calcium salt is calcium carbonate, calcium chloride or calcium hydroxide. 8. The method for producing phytin according to claim 1, wherein the yeast is a yeast that can consume lactic acid and is inoculated in a CSL or LSW medium in an amount of 1 to 20%. 9. The method for producing phytin according to claim 1, wherein the major ion of the precipitate is magnesium ion. 10. The method for producing phytin according to claim 1, wherein the fermentation is carried out batchwise. 11. The method for producing phytin according to claim 1, wherein the fermentation is performed in a semi-continuous manner. 12. The method for producing phytin according to claim 1, wherein the fermentation is carried out continuously.