JP2020183364A - Yeast protein-containing composition - Google Patents

Abstract

To provide a composition that contains nutrients such as protein and has health functionality such as the effects of inhibiting increases in blood glucose level or neutral fat.SOLUTION: The inventors have found that yeast (yeast protein) not only supplies protein but also has health functions, e.g. inhibiting increases in blood glucose or neutral fat.SELECTED DRAWING: None

Description

The present invention relates to a yeast protein-containing composition and a composition having an effect of suppressing an increase in blood glucose level or an effect of suppressing an increase in triglyceride.

With the development of a super-aging society, the importance of extending healthy life expectancy is increasing.
On the other hand, the food crisis caused by the increase in the world population and the low food self-sufficiency rate (especially protein) in Japan have become problems.

Under these circumstances, the concept of functional foods has been attracting attention from the concept of extending healthy life expectancy in everyday meals without relying on pharmaceuticals.
With the growing health consciousness, the number of foods to which food materials with health functionality are added is increasing year by year.
Foods for health care and foods with functional claims are attracting attention. Food materials having various health functionalities are known, and examples of functional foods that suppress an increase in blood glucose and triglyceride include indigestible kistrin (Patent Document 1). However, indigestible dextrin is a dietary fiber in which glucose is separated by allowing α-amylase and glucosidase to act on roasted dextrin derived from umorokoshi starch, and therefore contains almost no nutrients such as protein.

Further, it is known that yeast-derived protein is used as a substitute for soybean- and wheat-derived protein (Patent Document 2). These are supposed to be used as nutrients, and there was no concept of using them as health functional materials such as suppressing the rise of blood sugar and triglyceride.

Japanese Unexamined Patent Publication No. 2016-119861 Japanese Unexamined Patent Publication No. 2013-53083

An object of the present invention is to provide a composition containing nutrients such as protein and having health functionality such as an effect of suppressing an increase in blood glucose level or an effect of suppressing an increase in triglyceride.

The present inventors have found in diligent research that yeast (yeast protein) has not only protein supplementation but also health functions such as suppression of increase in blood glucose and triglyceride, and completed the present invention.

(1) A yeast protein-containing composition which is a cell wall-degrading enzyme-treated product of yeast cells or yeast residues.
(2) A yeast protein-containing composition for suppressing an increase in blood glucose level, which is a cell wall-degrading enzyme-treated product of yeast cells or yeast residues.
(3) A composition containing a mother protein for suppressing the increase in triglyceride in blood, which is a cell wall-degrading enzyme-treated product of yeast cells or yeast residues.
(4) A food containing the yeast protein-containing composition of (1) above, which suppresses the postprandial blood glucose level, moderates the postprandial blood glucose level rise, or raises the postprandial blood triglyceride level. Foods that are labeled as suppressing or slowing the rise of triglyceride in the blood after eating.
(5) A method for producing a yeast protein-containing composition, which comprises a step of treating yeast cells or yeast residues with a cell wall-degrading enzyme.

According to the present invention, a composition obtained by treating yeast cells (also referred to as "yeast residue" in the present invention) after extracting yeast cells, yeast extract, etc. with a cell wall-degrading enzyme is a protein and blood glucose as nutrients possessed by yeast. It is possible to provide a composition having health functionality such as a value increase suppressing effect or a neutral fat increase suppressing effect. In the present invention, a health functional effect was also found in yeast protein.

Relationship between enzyme concentration and viscosity / solubilization rate of reaction solution Relationship between enzyme reaction time and viscosity / solubilization rate (Brix) of reaction solution Results of molecular weight distribution analysis of yeast protein-containing composition Postprandial blood glucose suppression effect of yeast protein-containing composition Postprandial blood triglyceride inhibitory effect of yeast protein-containing composition Glucosidase inhibition of yeast protein-containing composition Lipase inhibition and bile acid adsorption test of yeast protein-containing composition

The yeast protein-containing composition of the present invention can be obtained by treating yeast cells or yeast residues with a cell wall-degrading enzyme.

Examples of the yeast used in the present invention include bacteria belonging to the genera such as saccharomyces, endomicopsis, saccharomycodes, nematospora, candida, tolulopsis, pretanomyces, and rhodotorula, or so-called brewer's yeast, baker's yeast, and sake yeast. Of these, Candida utilis or Saccharomyces cerevisiae, who have a lot of eating experience, are desired.

In the present invention, the yeast culture method is not limited, and a general method can be adopted. Generally, as a medium for culturing yeast, glucose, acetic acid, ethanol, glycerol, sugar honey, sulfite pulp waste liquid, etc. are used as carbon sources, and urea, ammonia, etc. are used as nitrogen sources. Ammonium sulfate, ammonium chloride, nitrate, etc. are used. The source of phosphate, potassium and magnesium may be ordinary industrial raw materials such as lime superphosphate, ammonium phosphate, potassium chloride, potassium hydroxide, magnesium sulfate and magnesium chloride, and other sources such as zinc, copper, manganese and iron ions. Add inorganic salt. Others can be cultured without using vitamins, amino acids, nucleic acid-related substances, etc., but these may be added. Organic substances such as corn stee bricker, casein, yeast extract, meat extract and peptone may be added.

Culturing conditions such as culturing temperature and pH can be applied without particular limitation, and may be set and cultivated according to the yeast strain to be used. Generally, the culture temperature is 20 to 40 ° C. and the pH is 3.0 to 9.0. The culture form may be either batch culture or continuous culture, and the conditions such as stirring and aeration during culture are not particularly limited, and a general method may be used.

In the present invention, yeast cells obtained by general culture of yeast can be used as they are. Yeast cells produced in the beer manufacturing process can also be used. It is also possible to use the yeast cell (yeast residue) that remains after extracting a useful component such as yeast extract from the yeast cell. The method for extracting yeast extract and useful components is not particularly limited, but yeast extract or yeast extract can be obtained by extracting yeast with one or more of hot water, acid / alkaline solution, autolysis, mechanical crushing, etc. Extracted useful components can be used.

In the present invention, the yeast cell wall-degrading enzyme is allowed to act on the yeast cells or yeast residues obtained by the above method. It is preferable to use glucanase as the yeast cell wall degrading enzyme. Furthermore, it is particularly preferable to use a glucanase that does not contain protease or has no protease activity. As long as the cell wall-degrading enzyme does not have protease activity, the cell wall-degrading enzyme may be allowed to act at a temperature and pH at which the protease in the enzyme preparation does not act, even if the protease is contained. Examples of enzymes having low or no protease activity include the cell wall degrading enzyme "Denateam GEL" (manufactured by Nagase ChemteX) derived from the genus Streptomyces, "Filtase BRX" (manufactured by DSM) derived from the genus Talaromyces, and the like. it can. When glucanase containing no protease is used, or when protease treatment is performed under conditions in which protease does not act, protease treatment may be performed after glucanase treatment. Any protease can be used.

The enzyme treatment conditions are not particularly limited. Depending on the enzyme used, those skilled in the art may have conditions that can be appropriately selected. The suspension concentration of yeast cells and the like, the enzyme concentration, etc. are arbitrary, but in general, the suspension concentration of yeast cells and the like is 5 to 20% by weight, and the addition concentration of glucanase is 0.001 to 10 with respect to yeast. %, preferably 0.01 to 0.5%, and the pH is preferably near neutral, but it may be within the pH range in which the enzyme acts. The reaction temperature is also preferably near the optimum temperature, and is not particularly limited. The enzyme reaction time is arbitrary. The enzyme reaction may be appropriately adjusted by those skilled in the art depending on the mode of use, such as when the product of the present invention is directly ingested or when it is mixed with other foods. For example, when the product of the present invention is directly ingested, the enzyme is added to yeast or yeast residue so as to be 0.3% by weight, and the mixture is reacted for 5 hours. This makes the yeast or yeast residue highly soluble in water and is easy to handle.

After the enzyme treatment, the component that dissolves in water and the component that does not dissolve in water may be separated, but they may not be separated and may be used as they are as the yeast protein-containing composition of the present invention.

The protein content of the present invention was calculated by the Kjeldahl method (calculated by multiplying the total amount of nitrogen by the nitrogen-protein conversion coefficient of 6.25). In the present invention, it is preferable that the composition contains 50% by weight or more. Other ingredients are optional, but may contain dietary fiber, vitamins and the like.

The obtained yeast enzyme-treated product may be dissolved in water or the like as it is and ingested as a beverage, or may be used or cooked in the form of a supplement by, for example, tableting, or mixed with general foods as a protein material. .. In addition, it may be used as a liquid seasoning by salting or concentrating. When it is dried, there is no limitation on the drying method. As a commonly used powdering method, a general powdering method such as spray drying, freeze drying, and drum drying can be used.

The amount and frequency of intake when ingested by humans differ depending on the form of intake, the age of the ingested person, the body weight, and the like. Usually, the composition of the present invention may be ingested in an amount of 1 to 40 g per day.

The invention of the present application will be specifically shown below, but the invention of the present application is not limited thereto.

The various measurement methods and the like in the present application are as follows.
<Viscosity measurement method>
Viscosity is 10% by weight, 50 ° C. and 7 using a b-type viscometer (TVB-10M, manufactured by Toki Sangyo Co., Ltd.).
The viscosity at 0 ° C. was measured.

<Measurement method of molecular weight distribution>
The molecular weight distribution of the soluble fraction of the enzyme-treated yeast protein was measured by HPLC analysis using gel filtration. Add 1 mL of ultrapure water to 25 mg of dried yeast protein composition obtained by drying enzyme-treated yeast protein, sufficiently vibrate and disperse with vortex, and then centrifuge (15,000 rpm, 3 minutes) to collect the supernatant. did. After filtering with a 0.45 μm filter, HPLC analysis was performed under the following conditions.
<HPLC conditions>
HPLC: Agilent 1200 System Mobile Phase: Ultrapure Water Column: YMC-Pack Diol-300 (5 μm, 300 × 8.0 mm ID, manufactured by Nacalai Tesque)
Column oven temperature: 30 ° C
Detector: RI
Flow velocity: 0.7 mL / min.
Inject amount: 50 μL

<Solubilization rate>
The solubilization rate is calculated by calculating the solid content of yeast protein and the yeast protein-soluble fraction (total solid content (%): W1, soluble fraction solid content (%): W2) by the atmospheric heating and drying method, and using the following formula. The solubilization rate was calculated.
Solubilization rate (%) = (100-W1) x W2 ÷ W1
(Pretreatment method)
5 g of 1 sample was collected, and 45 g of distilled water was added.
2 Stirrer for 30 minutes to disperse well.
3 The sample for total solid content measurement was sampled as it was.
4 For measuring the soluble solid content, 10 mL was weighed in a 15 mL centrifuge tube, centrifuged at 10,000 rpm (9,000 G) for 10 minutes, and the supernatant was collected and sampled.
(Solid content measurement method)
The total amount of yeast protein solids (%, W1) and soluble fraction solids (%, W2) were measured by a normal pressure heating and drying method at 105 ° C. for 4 hours with a sample amount of 2 g and sea sand used of 10 g.

(Physical properties of yeast protein-containing composition)
1 kg of dried bacterial cells of KR yeast for food (manufactured by Kojin Life Science Co., Ltd.) was suspended in water to adjust the bacterial cell concentration to 10% by weight, adjusted to 40 ° C. and pH 7.0, and then the cell wall degrading enzyme (cell wall degrading enzyme). "Glucanase GEL" manufactured by Nagase ChemteX Co., Ltd.) was added to the yeast so that the enzyme content was 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.3% by weight per yeast cell. It was added and reacted for 5 hours. After the reaction, the viscosity and solubilization rate of the 10% solution were measured, and the results shown in FIG. 1 were obtained. Note that FIG. 1 shows the results when KR yeast was used.

Yeast protein-containing compositions having different physical properties can be produced by changing the enzyme concentration as shown in FIG. 1, but can also be produced by changing the enzyme reaction time as shown below. FIG. 2 shows the results of measuring the viscosity and the supernatant Brix by adding 0.03% by weight of the enzyme to the yeast cells. In this test, the viscosity was measured using a tuning fork vibration rheometer RV-10000 (manufactured by A & D Co., Ltd.) at a sample concentration of 10% suspension at 50 ° C. and an amplitude of 1.0 mm. In addition, the Brix of the supernatant solution obtained by centrifuging the suspension at 15,000 rpm was measured as a guideline for the solubilization rate.
From the results shown in FIGS. 1 and 2, in order to produce a low-viscosity type yeast protein-containing composition (H form) in which the cell wall is highly decomposed, it is more suitable to increase the enzyme concentration and shorten the reaction time. It can also be produced by long-term enzyme treatment at a low concentration. In addition, when producing a highly viscous type yeast protein-containing composition (L-form) with low decomposition of the cell wall, there is a merit that the production can be completed in a short time by allowing a high concentration of enzyme to act, but to ensure physical properties. Since it is difficult to set the reaction time of the enzyme, it is more suitable to lower the enzyme concentration for stable production.
In this way, the enzyme concentration and the enzyme reaction time can be timely selected by the manufacturer according to the purpose.

FIG. 3 shows a part of the results of molecular weight distribution analysis of the obtained yeast protein-containing composition. The three peaks (1), (2), and (3) from the one with the largest molecular weight are characteristic. The H form is obtained by adding 0.3% by weight of the enzyme and reacting for 5 hours, and the L form is obtained by adding 0.03% by weight of the enzyme and reacting for 5 hours. As the enzyme condition becomes stronger, the peak of (1) decreases and the peak ratios of (2) and (3) increase.

(Production of yeast protein-containing composition)
An example of producing a yeast protein using KR yeast as the yeast and "Denateam GEL" (manufactured by Nagase ChemteX Corporation) as the glucanase is shown below. After suspending 2 kg of torula yeast in 18 kg of water and adjusting the pH to 7.0, 0.03% by weight (L-form of yeast protein-containing composition) or 0.3% by weight (yeast) of Denateam GEL with respect to KR yeast. It was added so as to be a protein-containing composition H form), and the mixture was stirred at 60 ° C. for 5 hours. The reaction solution was stirred at 90 ° C. or higher for 10 minutes to inactivate the enzyme, and then powdered by spray drying to obtain a yeast protein-containing composition. The protein content in the yeast protein-containing composition was 62% by weight as measured by the Kjeldahl method. Further, it was resuspended in water, and the water-soluble fraction and the water-insoluble fraction (high protein content fraction) were separated.

(Effect of suppressing postprandial blood glucose level of yeast protein-containing composition)
The efficacy test of the yeast protein-containing composition was carried out. KR yeast, yeast protein-containing composition L, yeast protein-containing composition H, pine fiber, and β-glucan-rich soybean flour were tested in healthy adult men and women according to the following protocol.

(Test method)
Step 1: Ingest a loaded diet (a food in which 75 g of glucose is dissolved in 200 ml of water) Step 2: Ingest a substance in which KR yeast is dispersed in the loaded diet of Step 1: Yeast protein-containing composition H in the loaded diet of Step 1. Body-dispersed intake Step 4: Ingestion of the yeast protein-containing composition H-form ingested with the water-insoluble fraction dispersed in the load diet of Step 1.

Hereinafter, in the same manner as above, each sample (yeast protein-containing composition L, pine fiber, β-glucan-rich soybean flour) was dispersed or dissolved and ingested.
* The rest period between steps is 2 days or more.

(Usage / Dose)
In the tests after step 2, the following volumes were dissolved or dispersed in the loaded diet of step 1 and ingested. The intake is shown in FIG.
Blood was collected from the fingertips with a lancet at 0 minutes, 30 minutes, 45 minutes, 60 minutes, 75 minutes, 90 minutes, and 120 minutes after ingestion, and the blood glucose level was measured.
ΔBlood glucose AUC% was calculated by the following formula.
Δ Blood glucose AUC% = (total area of blood glucose level up to 120 minutes after ingestion of 75 g glucose added to the test sample) ÷ (total area of blood glucose level up to 120 minutes after ingestion of 75 g glucose) × 100%

The results are shown in FIG.
Yeast itself, those with low degradation of the enzyme cell wall, and those with high degradation were all found to have an effect of suppressing the increase in postprandial blood glucose level.
In addition, the effect of suppressing the increase in blood glucose level was observed not only in the yeast protein-containing composition having an unwater-soluble fraction, but also in the protein-rich water-insoluble fraction.

(Suppressive effect of postprandial triglyceride)
KR yeast, yeast protein-containing composition L, yeast protein-containing composition H, pine fiber, and β-glucan-rich soybean flour were tested in healthy adult men and women according to the following protocol.

(Test food)
Test food 1 KR yeast 5g
Test food 2 Yeast protein composition L-form 3 g
Test food 3 Yeast protein composition H body 3 g
Test food 4 Yeast protein composition H-form insoluble fraction 2.4 g
Test food 5 Pine fiber 6g
Test food 6 β-glucan rich soy flour 3g

(Test method)
Step 1: 46.4g total fat meal menu + 200g water
Step 2: Step 1 meal menu + test food 1 suspended in 200 g of water Step 3: Step 1 meal menu + test food 2 suspended in 200 g Step 4: Step 1 meal menu + test Food 3 suspended in 200 g of water Step 5: Meal menu of step 1 + test food 4 suspended in 200 g of water Step 6: Meal menu of step 1 + test food 5 dissolved in 200 g of water Step 7: Meal menu of step 1 + test food 6 suspended in 200 g of water A rest period of 24 hours or more is provided between each step. The composition of the meal menu of 46.4 g of total fat is as follows: Frozen hamburger 1 Pieces (fat: 25.1 g)
2 butter rolls (fat: 16.0 g)
1 bag of french fries (100g) (fat: 5.3g)

Blood was collected from the fingertips with a lancet at 1, 2, 3, 4, 5, and 6 hours after ingestion, and the amount of triglyceride (TG) in the blood was measured.
The ΔTG value AUC% was calculated by the following formula.

ΔTG value AUC% = (high-fat diet + total area of blood TG up to 6 hours after ingestion of test sample) ÷ (total area of blood TG up to 6 hours after ingestion of high-fat diet) × 100%

The results of the efficacy test are shown in Fig. 5. Yeast itself and those with low degradation of the enzyme cell wall
All of the highly decomposed ones were found to have an effect of suppressing the increase in postprandial triglyceride.
In addition, the effect of suppressing the increase in triglyceride level was observed not only in the yeast protein-containing composition that had not been fractionated, but also in the protein-rich water-insoluble fraction. Note that 0.03% 5h and 0.3% 5h in the graph represent the enzyme addition concentration and the reaction time. The enzyme-treated yeast protein composition was found to have a stronger inhibitory effect on the increase of triglyceride due to the synergistic effect of the protein and the lysing component of the cell wall.

(Α-Glucosidase inhibitory effect of yeast protein-containing composition)
In order to elucidate the mechanism of action of the hypoglycemic effect of the yeast protein-containing composition, the α-amylase inhibitory effect and the α-glucosidase inhibitory effect were tested according to the following protocol.

(Α-Amylase inhibition)
<Enzyme solution>
A porcine pancreas-derived α-amylase (manufactured by SIGMA) dissolved in 0.1 M phosphate buffer (pH 6.9) containing 50 mM NaCl to 5 U / mL was used.

<Sample solution>
Acarbose (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.) was used as a positive control.
The sample solution used was dissolved or dispersed in 0.1 M phosphate buffer (pH 6.9) containing 50 mM NaCl as follows.
Yeast, yeast-containing composition, pine fiber: 100 mg / mL
Acarbose: 1 mg / mL

<Substrate solution>
0.1 M phosphate buffer with 1% soluble starch / 50 mM NaCl (pH 6.9)

<Operation procedure>
1 Mix 100 μL of sample solution and 100 μL of substrate solution.
2 Add 100 μL of the enzyme solution and soak at 37 ° C for exactly 10 minutes for mixing.
3 Add 1 mL of 0.5 mol / L acetic acid aqueous solution and stop the reaction.
4 Centrifuge at 15,000 rpm for 2 minutes and collect the supernatant.
5 Add 500 μL of 0.15% iodine-1.5% potassium iodide aqueous solution to 500 μL of the collected supernatant solution and mix.
6 Centrifuge at 15,000 rpm for 2 minutes.
7 Abs 700 nm is measured.
8 The residual starch concentration X was determined from the calibration curve.
Further, the enzyme solution was not added in the step 2, but the enzyme solution was added after the step 3, Abs 700 nm was calculated, and the starch concentration C was determined from the calibration curve.

The inhibitory activity was calculated by the following formula.
Inhibition rate (%) = 100- (1-C / X) x 100

(Α-Glucosidase inhibition)
<Crude enzyme solution>
As for the crude enzyme solution, 1.5 g of rat intestinal acetone powder (manufactured by SIGMA) was suspended in 15 mL of 0.1 M phosphate buffer (pH 7.0), homogenized in ice water, and then centrifuged, and the supernatant was further 0.1 M. The one diluted 2-fold with phosphate buffer (pH 7.0) was used.

<Sample solution>
Acarbose (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.) was used as a positive control.
The sample solution used was dissolved or dispersed in 0.1 M phosphate buffer (pH 7.0) as follows.
Yeast, yeast-containing composition, pine fiber: 100 mg / mL
Acarbose: 1 mg / mL

<Substrate solution>
4-Nitrophenyl-α-D-glucopyranoside (manufactured by Tokyo Chemical Industry Co., Ltd.) was used as a substrate, and the substrate was dissolved in 0.1 M phosphate buffer (pH 7.0) to a concentration of 10 mM.

<Operation procedure>
1 Mix 100 μL of sample solution and 100 μL of substrate solution.
2 Add 50 μL of enzyme solution and soak at 37 ° C for exactly 30 minutes for mixing.
3 Add 250 μL of 0.2 M aqueous sodium carbonate solution to stop the reaction. 4 Centrifuge at 15,000 rpm for 2 minutes, and collect the supernatant.
5 Measure Abs 415 nm of the supernatant.

The enzyme solution was not added in the step 2, the enzyme solution was added after the step 3, and the Abs 415 nm was calculated as blind, and the difference value from 5 was calculated, and the p-nitrophenol concentration X was obtained from the calibration curve.
A 0.1 M phosphate buffer (pH 7.0) was used instead of the substrate solution in the above step to determine the p-nitrophenol concentration C.

The inhibitory activity was calculated by the following formula.
Inhibition rate (%) = (1-X / C) x 100

The glucosidase inhibitory effect of the yeast and yeast protein-containing composition is shown in FIG.
Glucosidase inhibitory effect was observed in yeast and yeast protein-containing compositions, and glucosidase inhibition was presumed as one of the mechanisms of action of yeast and yeast protein-containing compositions to suppress the increase in postprandial blood glucose level.

(Lipase-suppressing effect of yeast protein-containing composition)
In order to elucidate the mechanism of action of the hypoglycemic effect of the yeast protein-containing composition, the lipase inhibitory effect and the bile acid adsorption test were tested according to the following protocol.

(Lipase inhibitory activity)
<Enzyme solution>
Pig pancreatic lipase (manufactured by SIGMA) dissolved in 0.1 M Tris buffer (0.1 M Tris-HCl buffer containing 0.1 M NaCl, pH 7.0) at a concentration of 1 mg / mL was used.

<Sample solution>
Orlistat (manufactured by Tokyo Chemical Industry Co., Ltd.) was used as a positive control.
The sample concentration was dissolved or dispersed in 0.1 M Tris buffer (0.1 M Tris-HCl buffer containing 0.1 M NaCl, pH 7.0) so as to be as follows.
Yeast, yeast-containing composition, pine fiber: 100 mg / mL
Orlistat: 0.1 mg / mL

<Substrate solution>
The substrate solution was prepared with the following composition and then homogenized by ultrasonic waves.
Olive oil (manufactured by Ajinomoto Co., Inc.): 2g
Soy lecithin (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.): 250 mg
Sodium cholic acid (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.): 100 mg
Female up to 50 mL with 0.1 M Tris buffer (0.1 M Tris-HCl buffer containing 0.1 M NaCl, pH 7.0)

<Operation procedure>
1 Mix 100 μL of sample solution and 100 μL of substrate solution.
2 Add 50 μL of the enzyme solution and soak and mix at 37 ° C for exactly 60 minutes.
3 After allowing to stand in boiling water for 2 minutes, add 500 μL of 0.2 M aqueous sodium carbonate solution to stop the reaction. Then, add 500 μL of 0.1 M Tris buffer to dilute.
4 Centrifuge at 15,000 rpm for 2 minutes and collect the supernatant.
5 Measure Abs 550 nm of the reacted supernatant according to the protocol of NEFA C-Test Wako (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.) In step 61, the same operation was performed using the enzyme solution deactivated in boiling water in advance. The value obtained by calculating Abs 550 nm was blinded to calculate the difference value from 5, and the free fatty acid concentration X in terms of oleic acid was obtained from the calibration curve.

Instead of the substrate solution in the above step, 0.1 M Tris buffer (0.1 M Tris-HCl buffer containing 0.1 M NaCl, pH 7.0) was used to determine the free fatty acid concentration C in terms of oleic acid.

The inhibitory activity was calculated by the following formula.

Inhibition rate (%) = (1-X / C) x 100

(Bile acid adsorption test)
<Sample>
Cholestyramine (manufactured by SIGMA-ALDRICH) was used as a positive control.
The amount of sample added is as follows.
Cholestyramine: 25 mg
Yeast, yeast-containing composition, pine fiber: 200 mg

<Operation procedure>
1 Weigh the sample into a 15 ml centrifuge tube and suspend in 5 ml of water.
2 Add 5 ml of 0.2 mM sodium cholic acid solution and suspend (use 0.2 M phosphate buffer (pH 6.8) to reach a final concentration of 0.1 mM sodium cholic acid / 0.1 M phosphate buffer).
3 Inversion mixing with a shaker (37 ° C, 2h, 100rpm).
42 Take 2 ml into Eppen and centrifuge (15000 rpm, 10 min).
5 The supernatant is collected and filtered through a filter (0.45 μm).
6 Abs 560 nm of Kamisei was measured according to the protocol of total bile acid-Test Wako (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.).
7 Using the phosphate buffer suspension of the sample (not containing sodium cholic acid), the one that was subjected to the same operation from step 3 onward was blinded, and the difference value from step 5 was calculated, and the free bile acid concentration was calculated from the calibration curve. X (mM) was determined.

The bile acid adsorption rate was calculated by the following formula.
Bile acid adsorption rate (%) = (1-X / 0.1) x 100

The lipase inhibitory effect of the yeast and yeast protein-containing composition is shown in FIG. Lipase inhibition was observed in both yeast and yeast protein-containing compositions. Furthermore, since the effects of yeast and yeast protein-containing compositions were also observed in the bile acid adsorption test, the yeast material was used as the mechanism of action of the postprandial neutral fat elevation inhibitory effect of yeast and yeast protein-containing compositions. It was speculated that one of the factors was that the adsorption of acids inhibited the formation of micelles in fats and oils, making lipase less effective.

Claims (5)

A yeast protein-containing composition which is a cell wall-degrading enzyme-treated product of yeast cells or yeast residues. A yeast protein-containing composition for suppressing an increase in blood glucose level, which is a cell wall-degrading enzyme-treated product of yeast cells or yeast residues. A yeast protein-containing composition for suppressing the rise of triglyceride in blood, which is a cell wall-degrading enzyme-treated product of yeast cells or yeast residues. A food containing the yeast protein-containing composition according to claim 1, which suppresses the rise in blood glucose level after meals or moderates the rise in blood glucose level after meals, or suppresses the rise in blood triglyceride after meals or after meals. Foods that claim to slow the rise of triglycerides in the blood. The method for producing a yeast protein-containing composition according to claim 1, which comprises a step of treating a yeast cell or a yeast residue with a cell wall-degrading enzyme.