Method for extracting animal feed, food raw material and yeast cell wall

Abstract

A feed for animals, which is a yeast cell wall extracted from yeast cultured using potato residues, the potato residues being residues obtained after starch is removed from potatoes, and wherein when the content of beta-glucan and chitin in the yeast cell wall is measured, the two conditions represented by the following mathematical formula (F1) and the following mathematical formula (F2) are satisfied, and when the content of beta-glucan and chitin in the yeast cell wall is measured, the content of beta-glucan and chitin in the yeast cell wall is measured by the following mathematical formula (F1) and the following mathematical formula (F2). GYPD / G > 1... (F1) CYPD / C < 1... (F2) G: the content (mass%) of beta-glucan in the yeast cell wall C: the content ([mu] mol / mg) of chitin in the yeast cell wall GYPD: the content (mass%) of beta-glucan in the yeast cell wall extracted from yeast cultured using a YPD culture medium CYPD: The content ([mu] mol / mg) of chitin in yeast cell walls extracted from yeast cultured using a YPD medium.

Description

Technical Field The invention relates to a method for extracting animal feed, food raw materials and yeast cell walls. Background Art As a method of utilizing the residue after starch is extracted from tubers, animal feed and the like have been studied. For example, Patent Document 1 describes a method for producing animal or fish feed by fermenting cassava residue with amylolytic enzyme and microorganisms. Prior art literature Patent Literature Patent Document 1: Japanese Patent Application Publication No. 2017-035005 Summary of the invention Problems to be solved by the invention According to the method for producing animal or fish feed described in Patent Document 1, animal feed can be obtained by utilizing cassava residues and the like which have not been used in the past. However, Patent Document 1 does not disclose any knowledge regarding the active ingredients in the animal feed. The object of the present invention is to provide an extraction method of animal feed, food raw materials and yeast cell walls that can regulate the proliferation of immune cells or immunity. Means for solving problems According to one embodiment of the present invention, the following animal feed, food raw material, and method for extracting yeast cell wall are provided.

[0001] Animal feed, which is yeast cell wall extracted from yeast cultured with potato residues, wherein the potato residues are the residues after starch is taken out of potatoes, When the contents of β-glucan and chitin in the yeast cell wall are measured, the two conditions shown in the following mathematical formula (F1) and the following mathematical formula (F2) are satisfied. G YPD / G>1···(F1) C YPD / C<1···(F2) G: The content of β-glucan in the yeast cell wall (mass %) C: Chitin content in the yeast cell wall (μmol / mg) G YPD : Content of β-glucan in yeast cell wall extracted from yeast cultured in YPD medium (mass %) C YPD : The chitin content (μmol / mg) in the yeast cell wall extracted from yeast cultured in YPD medium.

[0002] The animal feed according to [1], wherein The G YPD / G value is 1.19 or more, The C YPD The value of / C is 0.98 or less.

[0003] The animal feed according to [1] or [2], wherein The tuber is at least one selected from potato, sweet potato, Jerusalem artichoke, taro, taro, purple taro, konjac, yam, cassava, Chinese yam and Japanese yam.

[0004] The animal feed according to any one of [1] to [3], The animal is at least one selected from the group consisting of dairy cows, beef cattle, sheep, pigs, chickens and fish.

[0005] The food raw material is yeast cell wall extracted from yeast cultured with potato residue, wherein the potato residue is the residue after starch is taken out from potatoes, When the contents of β-glucan and chitin in the yeast cell wall are measured, the two conditions shown in the following mathematical formula (F1) and the following mathematical formula (F2) are satisfied. G YPD / G>1···(F1) C YPD / C<1···(F2) G: The content of β-glucan in the yeast cell wall (mass %) C: Chitin content in the yeast cell wall (μmol / mg) G YPD : Content of β-glucan in yeast cell wall extracted from yeast cultured in YPD medium (mass %) C YPD : The chitin content (μmol / mg) in the yeast cell wall extracted from yeast cultured in YPD medium.

[0006] A method for extracting yeast cell wall, comprising: The process of subjecting potato residue to ethanol fermentation with starch decomposing enzyme and yeast, and culturing the yeast to obtain ethanol fermentation product, wherein the potato residue is the residue after starch is taken out from the potato; a step of separating the yeast from the ethanol fermentation product; and A step of extracting yeast cell walls satisfying the following condition 1 from the yeast; (Condition 1) When the contents of β-glucan and chitin in the yeast cell wall are measured, the two conditions shown in the following mathematical formula (F1) and the following mathematical formula (F2) are satisfied. G YPD / G>1···(F1) C YPD / C<1···(F2) G: The content of β-glucan in the yeast cell wall (mass %) C: Chitin content in the yeast cell wall (μmol / mg) G YPD : Content of β-glucan in yeast cell wall extracted from yeast cultured in YPD medium (mass %) C YPD : The chitin content (μmol / mg) in the yeast cell wall extracted from yeast cultured in YPD medium. According to one embodiment of the present invention, a method for extracting animal feed, food raw materials, and yeast cell walls that can regulate the proliferation of immune cells or immunity can be provided. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 : is a graph showing the β-glucan content in the yeast cell walls (CP and YPD) obtained in Example 1 and Comparative Example 1. Figure 2 : is a graph showing the β-glucan content in the yeast cell walls (SP and YPD) obtained in Example 2 and Comparative Example 2. Figure 3 : is a graph showing the chitin content in the yeast cell walls (CP and YPD) obtained in Example 1 and Comparative Example 1. Figure 4 : is a graph showing the chitin content in the yeast cell walls (SP and YPD) obtained in Example 2 and Comparative Example 2. Figure 5 : This is a diagram showing the results of cell proliferation when peripheral blood mononuclear cells (PBMC) were cultured using the yeast cell wall (Pichia kudriavzevii) obtained in Example 1 and reagents (Concanavalin A and Zymosan). Figure 6 : It is a figure which shows the evaluation result of the influence on cytokines (tumor necrosis factor, TNFa) when the yeast cell wall (Pichia kudriavzevii) obtained in Example 1 and the reagent of yeast cell wall (Zymosan) were used. Figure 7 : It is a figure which shows the evaluation result of the influence on cytokines (interleukin-1β, IL-1b) when the yeast cell wall (Pichia kudriavzevii) obtained in Example 1 and the reagent of yeast cell wall (Zymosan) were used. DETAILED DESCRIPTION [Animal feed] First, the animal feed of the present embodiment will be described. The animal feed of the present embodiment is an animal feed obtained by extracting yeast cell walls from yeast cultured using yam residues, which are residues after starch is extracted from yam. Furthermore, when measuring the content of β-glucan and chitin in the yeast cell wall, the two conditions shown in the following mathematical formula (F1) and the following mathematical formula (F2) must be satisfied. G YPD / G>1···(F1) C YPD / C<1···(F2) G: The content of β-glucan in the yeast cell wall (mass %) C: Chitin content in the yeast cell wall (μmol / mg) G YPD : Content of β-glucan in yeast cell wall extracted from yeast cultured in YPD medium (mass %) C YPD : The chitin content (μmol / mg) in the yeast cell wall extracted from yeast cultured in YPD medium. G YPD When the value of G is less than 1, the proliferation effect of immune cells in the animal feed becomes insufficient. YPD / G is preferably 1.05 or more, more preferably 1.1 or more, and particularly preferably 1.19 or more. YPD There is no particular upper limit on the value of / G. YPD The value of / G may be 10 or less, or 5 or less. It should be noted that C YPD The value of / C is preferably 0.98 or less, more preferably 0.94 or less. YPD There is no particular restriction on the lower limit of / C. YPD The value of / C may be 0.1 or more, or 0.2 or more. As G YPD / G value and C YPD The method for adjusting the value of / C to fall within the above range includes, for example, the method for extracting the yeast cell wall of the present embodiment described later. The animal feed of the present embodiment is preferably used for at least one animal selected from the group consisting of dairy cows, beef cattle, sheep, pigs, chickens, and fish. The potato residue used in the present embodiment is the residue after starch is extracted from potatoes. Examples of the tubers include potato, sweet potato, Jerusalem artichoke, taro, taro, purple taro, konjac, yam, cassava, Chinese yam, Japanese yam, etc. Among them, sweet potato or cassava is preferred from the viewpoint of raw material availability. The yeast used in the present embodiment is not particularly limited, and examples of the yeast include Pichia kudriavzevii, Saccharomyces cerevisiae, and Kluyveromyces marxianus. [Extraction method of yeast cell wall] Next, the method for extracting yeast cell walls of the present embodiment is described. The method for extracting yeast cell walls of the present embodiment is a method having the following steps: a step of performing ethanol fermentation on potato residues using amylolytic enzymes and yeast, culturing the yeast to obtain an ethanol fermentation product (hereinafter also referred to as the "first step"), a step of separating the yeast from the ethanol fermentation product (hereinafter also referred to as the "second step"), and a step of extracting yeast cell walls satisfying the following condition 1 from the yeast (hereinafter also referred to as the "third step"), wherein the potato residues are residues after starch is removed from potatoes. (Condition 1) When the contents of β-glucan and chitin in the yeast cell wall are measured, the two conditions represented by the above mathematical formula (F1) and the above mathematical formula (F2) are satisfied. According to the method for extracting yeast cell walls of the present embodiment, the animal feed of the present embodiment described above can be efficiently produced. In the first step, first, potato residues are prepared as residues after starch is removed from potatoes. Yams are as described above. The moisture content of the potato residue is not particularly limited, but from the viewpoint of saccharification and fermentation, the moisture content of the potato residue is preferably 30% by mass or more. As the potato residue, dried potato residue can be used. When using the dried potato residue, water can be added to adjust the moisture content of the potato residue. In addition, the dried potato residue can also be mixed with potato residue with high moisture content after starch is squeezed out. In the first step, the potato residue is then subjected to ethanol fermentation using amylolytic enzymes and yeast, and the yeast is cultured to obtain an ethanol fermentation product. The potato residue can be directly subjected to fermentation treatment. In order to gelatinize the starch remaining in the potato residue, it is preferably subjected to heat treatment before the fermentation treatment. As heating conditions, the heating temperature is preferably above 60°C, and the heating time is preferably above 5 minutes. If the heating temperature is above 60°C, the gelatinization efficiency of the starch remaining in the potato residue can be improved. The heating time is not particularly limited and can be appropriately set in consideration of the heating temperature. The amylolytic enzyme treatment and the yeast fermentation treatment may be performed separately or simultaneously. Yeast is as described above. The amylolytic enzyme is not particularly limited as long as it is an enzyme that decomposes starch. Examples of the amylolytic enzyme include α-amylase and saccharifying enzyme. In addition, in order to reduce the viscosity of the potato residue, in addition to the amylolytic enzyme, pectinase or cellulase may also be used in combination. The amount of α-amylase added is preferably 9×10 -5 Unit (hereinafter also referred to as "U") is greater than or equal to 600U and less than or equal to 8×10 -3 The amount of saccharifying enzyme is preferably 3×10 -4 U or more and 200 U or less, more preferably 3×10 -2 U or more and 0.2U or less. When cellulase is used, the amount of cellulase added is preferably 1×10 -4 When pectinase is used, the amount of pectinase added is preferably 1×10 -3 U or more and 1000 U or less, more preferably 1×10 -1 More than U and less than 1U. In the second step, yeast is separated from the ethanol fermentation product. Liquid components such as ethanol contained in the ethanol fermentation product can be removed by at least one of a method of squeezing the ethanol fermentation product and a method of heating the ethanol fermentation product. If the liquid component is removed from the ethanol fermentation, the yeast can be isolated. In the third step, yeast cell walls are extracted from the yeast. In order to extract the yeast cell wall, it is preferred to first culture the yeast at a temperature of 50° C. for 10 hours or longer to cause autolysis, and then grind the yeast to obtain a grinded product. The pulverization method may be dry pulverization or wet pulverization. Examples of pulverization apparatus include a bead mill, a ball mill, and a colloid mill. Next, the obtained pulverized product is washed with ethanol or the like, and then the solid content is precipitated, whereby the yeast cell walls can be fractionated and extracted. As described above, yeast cell walls satisfying the aforementioned condition 1 can be extracted. [Food raw materials] Next, the food raw material of this embodiment is described. The food raw material of this embodiment is the same as the animal feed of the above-mentioned embodiment. As described above, the animal feed of this embodiment can regulate the proliferation of immune cells or immunity. Therefore, it can be expected to have health-enhancing effects as a food material. Example Next, the present invention will be described in further detail with reference to Examples and Comparative Examples, but the present invention is not limited to these Examples at all. [Example 1] 20 g of dried cassava residue and 80 g of pure water were mixed to obtain cassava residue having a water content of 80% by mass. 8 to 10 units of α-amylase were added to the obtained cassava residue based on 1 g of the solid content of the cassava residue, and the mixture was heated at 30 to 100° C. for 0.4 to 1 hour and then cooled to obtain a culture medium. An enzyme mixture (1.28 units of saccharifying enzyme, 4.0 units of pectinase, and 0.46 units of cellulase) and a nitrogen source (yeast extract + peptone) are added to the obtained culture medium, and then Pichia kudriavzevii is inoculated as a yeast. The yeast is cultured under the conditions of a temperature of 30 to 40°C, a stirring speed of 50 to 150 rpm, and 24 to 40 hours to obtain an ethanol fermentation product. Next, the obtained ethanol fermentation product is squeezed to remove the liquid containing ethanol to separate the yeast. The separated yeast is then autolyzed, pulverized using a bead mill, washed with ethanol, and then the solid content is precipitated to fractionate the yeast cell walls and extract the yeast cell walls. [Example 2] A culture medium and yeast cell walls were obtained in the same manner as in Example 1 except that 20 g of dried sweet potato residues were used instead of 20 g of dried cassava residues. [Comparative Example 1] Yeast cell walls were obtained in the same manner as in Example 1 except that YPD medium was used as the medium. [Comparative Example 2] Yeast cell walls were obtained in the same manner as in Example 2 except that YPD medium was used as the medium. [Determination of β-glucan and chitin content in yeast cell walls] The contents of β-glucan and chitin in the yeast cell walls obtained in Example 1, Example 2, Comparative Example 1 and Comparative Example 2 were measured as follows. β-glucan was measured using Enzymatic yeast beta glucan assay procedure (K-EBHLG) manufactured by Megazyme. Chitin was measured according to the following references 1 and 2 using the MBTH (3-methyl-benzothiazolinonehydrazone hydrochloride) method. Reference 1: Tsuji A, Kinoshita T, Hoshino M. 1969. Analytical chemicalstudies on amino sugars. II. Determination of hexosamines using 3-methyl-2-benzothiazolone hydrazone hydrochloride. Chem Pharm Bull (Tokyo) 17: 1505-1510. Reference 2: Tsuji A, Kinoshita T, Hoshino M. 1969. Microdetermination of hexosamines. Chem Pharm Bull (Tokyo) 17: 217-218. The contents of β-glucan in the yeast cell walls obtained in Example 1 and Comparative Example 1 are shown in Figure 1 In addition, the contents of β-glucan in the yeast cell walls obtained in Example 2 and Comparative Example 2 are shown in Figure 2 . The chitin contents in the yeast cell walls obtained in Example 1 and Comparative Example 1 are shown in Figure 3 The chitin content in the yeast cell wall obtained in Example 2 and Comparative Example 2 is shown in Figure 4 . It should be explained that Figure 1 to Figure 4 In the examples, Example 1 is labeled as CP because it uses a culture medium made of cassava residues, and Example 2 is labeled as SP because it uses a culture medium made of sweet potato residues. In addition, Comparative Examples 1 and 2 are labeled as YPD because they use YPD culture medium. also, Figure 1 , Figure 2 and Figure 4 The results are marked with error bars. Figure 3 In the figure, the first test result is recorded on the left, and the second test result is recorded on the right. Depend on Figure 1 and Figure 3The results shown in the figure confirmed that in the yeast cell wall obtained in Example 1, G YPD / G value exceeds 1, C YPD The value of / C is less than 1. In addition, by Figure 2 and Figure 4 The results shown in the figure confirmed that in the yeast cell wall obtained in Example 2, G YPD / G value exceeds 1, C YPD The value of / C is less than 1. [Evaluation of the culture of peripheral blood mononuclear cells] The effects of the yeast cell wall obtained in Example 1, Concanavalin A (manufactured by FUJIFILM Wako Pure Chemical Industries, Ltd.), and Zymosan (manufactured by FUJIFILM Wako Pure Chemical Industries, Ltd.) on the cultivation of peripheral blood mononuclear cells (hereinafter also referred to as "PBMC") were evaluated as follows. That is, PBMCs were separated from bovine blood and the cultured products were used as samples. Yeast cell walls and reagents were added to the samples, and PBMCs were proliferated under the conditions of 37°C and 72 hours. Then, Cell Counting Kit 8 (manufactured by Dojin Chemical Research Institute Co., Ltd.) was added to the proliferated samples, and the absorbance at 450nm (A450) was measured to evaluate the cultivation of PBMCs. The results are shown in Figure 5 It should be noted that Figure 5 In the examples, since Pichia kudriavzevii was used as yeast in Example 1, it was labeled as P. kudriavzevii, and the sample in which yeast cell wall and reagent were not added was labeled as control. In addition, the abbreviations of the reagent names were labeled. It should be noted that the abbreviations are as follows. ConA: Concanavalin A (3 μg / mL) Zymosan: β-Glucan Standard (100ug / mL) CP: Cassava residue Depend on Figure 5 The results shown confirmed that the yeast cell wall obtained in Example 1 had a significantly higher PBMC proliferation effect than Zymosan, a yeast cell wall reagent. It was also confirmed that the yeast cell wall obtained in Example 1 had a higher PBMC proliferation effect than Concanavalin A, an immune cell activating reagent. [Evaluation of the effects of cytokines] The effects of the yeast cell wall and Zymosan (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) obtained in Example 1 on cytokines were evaluated as follows. In addition, tumor necrosis factor (TNFα) and interleukin-1β (IL-1b) were evaluated as cytokines. Specifically, yeast cell walls and Zymosan were added to PBMCs separated from bovine blood, and the cells were stimulated at 37°C for 6 hours. After RNA was extracted, the gene expression levels of each cytokine relative to the standard gene, i.e., β-actin, were measured by quantitative PCR to evaluate the effect of yeast cell walls on cytokine production. The results of the evaluation of the effect on tumor necrosis factor (TNFα) when the yeast cell wall and the yeast cell wall reagent obtained in Example 1 were used are shown in FIG. Figure 6 . The results of the evaluation of the effect on interleukin-1β (IL-1b) when the yeast cell wall and the yeast cell wall reagent obtained in Example 1 were used are shown in FIG. Figure 7 . It should be noted that Figure 6 and Figure 7 In Example 1, since Pichia kudriavzevii was used as yeast, it was labeled as Pichia kudriavzevii (P. kudriavzevii) and the name of the labeling agent Zymosan. In addition, the example in which yeast cell wall was not used was labeled as a control. Depend on Figure 6 and Figure 7 The results shown confirmed that the yeast cell wall obtained in Example 1 can suppress gene expression compared with Zymosan, which is a yeast cell wall agent. In addition, it was also confirmed that the yeast cell wall obtained in Example 1 can suppress gene expression compared with the case where no yeast cell wall was used (control). Therefore, it was confirmed that the yeast cell wall obtained in Example 1 can regulate immunity.

Claims

1. Animal feed, which is yeast cell walls extracted from yeast cultured using potato residues, wherein the potato residues are the residues after starch is taken out of potatoes, wherein: When the contents of β-glucan and chitin in the yeast cell wall are measured, the two conditions shown in the following mathematical formula (F1) and the following mathematical formula (F2) are satisfied. G YPD / G>1···(F1) C YPD / C<1···(F2) G: The content of β-glucan in the yeast cell wall (mass %) C: Chitin content in the yeast cell wall (μmol / mg) G YPD : Content of β-glucan in yeast cell wall extracted from yeast cultured in YPD medium (mass %) C YPD : The chitin content (μmol / mg) in the yeast cell wall extracted from yeast cultured in YPD medium.

2. The animal feed according to claim 1, wherein The G YPD / G value is 1.19 or more, The C YPD The value of / C is 0.98 or less.

3. The animal feed according to claim 1 or 2, wherein: The tuber is at least one selected from potato, sweet potato, Jerusalem artichoke, taro, taro, purple taro, konjac, yam, cassava, Chinese yam and Japanese yam.

4. The animal feed according to claim 1 or 2, wherein: The animal is at least one selected from the group consisting of dairy cows, beef cattle, sheep, pigs, chickens and fish.

5. A food raw material, which is a yeast cell wall extracted from yeast cultured using potato residues, wherein the potato residues are the residues after starch is taken out of potatoes, wherein: When the contents of β-glucan and chitin in the yeast cell wall are measured, the two conditions shown in the following mathematical formula (F1) and the following mathematical formula (F2) are satisfied. G YPD / G>1···(F1) C YPD / C<1···(F2) G: The content of β-glucan in the yeast cell wall (mass %) C: Chitin content in the yeast cell wall (μmol / mg) G YPD : Content of β-glucan in yeast cell wall extracted from yeast cultured in YPD medium (mass %) C YPD : The chitin content (μmol / mg) in the yeast cell wall extracted from yeast cultured in YPD medium.

6. A method for extracting yeast cell walls, comprising: The process of subjecting potato residue to ethanol fermentation with starch decomposing enzyme and yeast, and culturing the yeast to obtain ethanol fermentation product, wherein the potato residue is the residue after starch is taken out from the potato; a step of separating the yeast from the ethanol fermentation product; and A step of extracting yeast cell walls satisfying the following condition 1 from the yeast; (Condition 1) When the contents of β-glucan and chitin in the yeast cell wall are measured, the two conditions shown in the following mathematical formula (F1) and the following mathematical formula (F2) are satisfied. G YPD / G>1···(F1) C YPD / C<1···(F2) G: The content of β-glucan in the yeast cell wall (mass %) C: Chitin content in the yeast cell wall (μmol / mg) G YPD : Content of β-glucan in yeast cell wall extracted from yeast cultured in YPD medium (mass %) C YPD : The chitin content (μmol / mg) in the yeast cell wall extracted from yeast cultured in YPD medium.

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