JP2006271377A - Enzymolysis product of animal liver and food containing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a food having lipid and carbohydrate metabolism ameliorating effect based on new knowledge that the enzymolysis product of pig liver is effective to ameliorate lipid and carbohydrate metabolism. <P>SOLUTION: This food having lipid and carbohydrate metabolism ameliorating effect is obtained by mixing other additive with liver powder containing much adjusted vitamin B, mineral (iron, zinc and copper) and nucleic acid and having amino acid score of 100, or an amino acid mixture with amino acid composition equivalent to the liver powder, or a mixture of glycin alanine. The liver powder is obtained by crushing pork liver, making the product react at optimum PH and 45°C for about 4 h using protease, extracting the product with pressure followed by concentrating with centrifugal separation, and adjusting the molecular weight of the product to 50-1,000 with extra filtration. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a food containing an enzymatic degradation mixture of animal liver, and more particularly to a food containing an enzymatic degradation mixture of pig liver or an equivalent mixture and having an effect of improving lipid metabolism and sugar metabolism.

  In recent years, lifestyle-related diseases such as diabetes, hypertension, and hyperlipidemia continue to increase due to westernization of eating habits and changes in living environment. Obesity is said to be a major cause of these lifestyle-related diseases, and extensive research and development have been conducted on its prevention methods and solutions.

  So far, the effectiveness of plant proteins and peptides (improvement of lipid metabolism, etc.) has been reported in the field of food nutrition. However, some studies on the functionality of meat and livestock by-products are limited. By the way, the liver is a food with a very high nutritional value that contains proteins, vitamins, minerals and the like in a well-balanced manner, and attention has been paid to this point, and the development of foods using this nutritional value has been conventionally performed.

  However, the utility value as food was low due to problems such as texture and smell. In order to solve this point, conventionally, as a deodorizing and deodorizing method of the lever, there has been proposed a hot water treatment using bran or adding catechin (see Patent Document 1).

In addition, a paste-like study has been conducted as a method for improving the texture. Furthermore, the invention has also been devised to improve taste defects by means of a hydrolysis technique using enzymes.
JP 2000-32955 A (see paragraph 0011)

  As described above, the “food texture and odor”, which are the secondary functions of food in pig liver, are being improved by the conventional technology, but the study with a view to the tertiary function (biological regulation function) of food Conventionally, it has not been performed sufficiently. Conventionally, an enzyme degradation product of pig liver has been present, but only a role of nutritional supplement was expected as a nutrient and vitamin source.

  The inventors of the present invention have been intensively researching the tertiary function (bioregulatory function) exhibited by the enzyme product of porcine liver as food. The enzyme product of porcine liver improves lipid and sugar metabolism. We obtained knowledge that it is effective. Therefore, an object of the present invention is to realize a food containing an enzymatic degradation mixture of animal liver having a molecular weight of 50 to 1000 having an effect of improving lipid and sugar metabolism based on this finding.

  Furthermore, the present invention searches for substances that contribute to the suppression of lipid accumulation among the enzymatic degradation products of pig liver, and therefore does not necessarily add substances derived from pig liver but corresponding synthetic and / or natural products. Thus, it is an object to realize a food having an effect of improving lipid and sugar metabolism.

  In order to solve the above-mentioned problems, the present invention provides an enzymatic degradation mixture of animal liver obtained by concentrating components obtained by enzymatic treatment of animal liver and improving lipid metabolism and sugar metabolism.

  The enzymatic degradation mixture is an enzymatic degradation mixture having a molecular weight of 50 to 1000 containing amino acids, water-soluble vitamins, minerals, nucleic acids, and peptides.

  The enzyme degradation mixture is prepared by pulverizing pork liver, reacting with protease at 45 ° C. at an optimum pH, extracting with pressure, concentrating, and adjusting the molecular weight to 50 to 1000 liver powder by ultrafiltration. Features.

  In order to solve the above-mentioned problems, the present invention pulverizes pork liver, reacts with protease at 45 ° C. at an optimum pH, extracts and concentrates under pressure, and adjusts the molecular weight to 50 to 1000 lever powder by ultrafiltration. There is provided an enzymatic degradation mixture comprising an enzymatic degradation mixture comprising a synthetic and / or natural product-derived amino acid mixture having an amino acid composition equivalent to that of an animal liver enzymatic degradation mixture.

  It should be noted that the amino acid composition equivalent to the above-mentioned animal liver enzymatic degradation mixture is ± 2% of the content of each amino acid compared to the content% indicated by the amino acid composition of the porcine liver enzymatic degradation product described in FIG. Preferably, each amino acid content% configured with a width of ± 1% refers to the amino acid composition. For example, if it is glycine content, it may be in the range of 1.05 to 5.05%, preferably 2.05 to 4.05% is desirable, and if it is alanine content, 1.48 to 5% It may be in the range of .48%, preferably in the range of 2.48 to 4.48%, and may be in a range having the same range for other amino acids.

  In order to solve the above-mentioned problems, the present invention provides an enzymatic degradation mixture characterized by improving glycine or alanine or a mixture of glycine and alanine and improving lipid metabolism and sugar metabolism.

  In order to solve the above-mentioned problems, the present invention provides a food containing any of the above enzymatic degradation mixtures.

  The food containing the enzyme degradation product of the animal liver according to the present invention is effective in improving lipid and sugar metabolism, and the liver enzyme degradation product can be used for staple foods, beverages, etc. to be eaten daily. Prevents lifestyle-related diseases such as hyperlipidemia, and has a great effect on improvement.

  Furthermore, the present invention adds a synthetic and / or natural product-derived substance equivalent to a pig liver-derived substance obtained as a result of searching for a substance that contributes to lipid accumulation suppression among pig liver enzyme degradation products. Thus, a food having an effect of improving lipid and sugar metabolism can be obtained.

  BEST MODE FOR CARRYING OUT THE INVENTION The best mode for carrying out a food containing an animal liver enzyme degradation mixture according to the present invention will be described below based on test examples and examples.

  The inventors of the present invention concentrated an enzyme-treated component of pork liver and further ultrafiltered to obtain an enzyme-degraded mixture containing amino acids as a main component. This enzymatic degradation mixture contains amino acids, water-soluble vitamins, minerals, nucleic acids, peptides and the like.

  When this enzymatic degradation mixture was administered to animals (rats) for a long time, it was confirmed that it was effective in improving lipid and sugar metabolism. In short, as a result of animal experiments, it was confirmed that the enzyme degradation mixture derived from animal liver was effective in improving lipid and sugar metabolism by long-term intake of the liver.

  And it concentrates the component which processed the pig liver with an enzyme, and then ultrafilters. Depending on the degree of this filtration, the size of the fraction molecular weight of the resulting enzymatic degradation mixture varies, but the molecular weight is 50-1000, In particular, it has been found that the enzymatic degradation mixture of pig liver having a molecular weight of 100 to 1000 has an extremely large effect of improving lipid and sugar metabolism.

  The present invention is based on the knowledge that such an enzyme degradation product of pig liver is effective in improving lipid and sugar metabolism, and has a molecular weight of 1000 or less, particularly 50 to It is a food containing 1000 porcine liver enzymatic degradation mixture.

  The animal liver enzymatic decomposition mixture of the present invention is specifically prepared as a liver powder, and the adjustment is performed as follows. Pork liver is crushed as a raw material, reacted with protease (for example, promain, chymotrypsin, papain, etc.) at an optimum pH of about 45 degrees for 4 hours, extracted under pressure, and concentrated by centrifugation.

  Further, the concentrate is adjusted to a molecular weight of 3000 or less by primary membrane filtration, and further adjusted to a molecular weight of 1000 or less, particularly 50 to 1000 by secondary membrane filtration, and pulverized. The prepared liver powder contains a large amount of vitamin B group, mineral (iron, zinc, copper, etc.) and nucleic acid, and the amino acid score is 100.

(Test Example 1)
Overview:
In order to demonstrate that the animal liver enzyme degradation mixture of the present invention obtained as a liver powder by the above-mentioned adjustment has an effect of improving lipid and sugar metabolism, as shown in Table 1, swine liver enzyme degradation product administration group (Lever group) and non-administration group (control group) were measured and evaluated for lipid metabolism improvement effect and glucose metabolism improvement effect.

  The porcine liver liver hydrolyzate administration group (lever group) is a case where two test foods A and B (lever group in Table 1) containing liver powder are administered to animals, and the non-administration group (control group) ) Is the case where the test meal C ("control group" in Table 1) containing no liver powder was administered to the animals.

Test meal:
Here, the mixed feeds of the test meals A and B are as shown in Table 1, and the mixed feeds other than the liver powder are common to each other. However, the properties of the lever powder contained in each of the test meals A and B are different from each other. That is, the fractional molecular weight of the test meal A is 50 to 1000, and the fractional molecular weight of the test meal B is 1000 to 100,000. This fractionated molecular weight is adjusted according to the degree of ultrafiltration as described above.

Test conditions:
The test conditions are as follows. In this administration test, type 2 diabetes model rat OLETF (Otsuka Long-Evans Tokyo Fatty) male 18 weeks old (n = 14) was purchased, and oral glucose tolerance test (2.0 g glucose / kg b. w.).

  Based on this result, the group was divided so that the constitution of glucose metabolism was not biased, and the intake of the test meal was started from the age of 20 weeks. Thereafter, blood sampling was performed three times every 4 weeks, and at the age of 34 weeks. Collected feces.

  For measurement and evaluation of lipid metabolism improvement effect and glucose metabolism improvement effect, specifically, body weight (g), blood cholesterol (mg / dl), free fatty acid, neutral fat, blood glucose level (mg / dl), plasma insulin level (pg / ml) and leptin concentration (pg / ml) were measured by conventional methods.

  For feces, dry fecal mass (g / day), fecal fat mass (mg / day), fat digestibility (%), acidic sterol (μmol / l), neutral sterol (μmol / l) are measured. did.

Test results and discussion:
As a test result, the change in body weight (g) during the period was, as shown in FIG. 1, the swine liver enzyme digested product administration group (lever from the 8th week (28 weeks in FIG. 1)) intake of the swine liver enzyme digest. Group) and a non-administration group (control group), a significant difference was observed between the swine liver enzyme degradation product administration group and the non-administration group. In addition, the meaning of p <0.05 in FIGS. 1-7 has suggested that there exists a significant difference as a result of t test.

  As shown in FIG. 2, the changes in various markers due to the intake of the test meal were as follows. The fasting blood glucose level (mg / dl) was determined from the 4th week (24 weeks of age in FIG. 2) of swine liver enzyme degradation product intake. Significant differences were observed between the liver enzyme degradation product administration group (left bar in the bar graph of FIG. 2) and the non-administration group (right bar of the bar graph in FIG. 2). However, no significant difference was observed in the insulin level (pg / ml).

  As shown in FIG. 3, the cholesterol level (mg / dl) is determined from the swine liver enzyme degradation product administration group (24 weeks of age in FIG. 3) from the pig liver enzyme degradation product administration group (left side of the bar graph in FIG. 3). A significant difference was observed between the non-administered group (the bar on the right side of the bar graph in FIG. 3). However, there was no significant difference in NEFA value (mEq / l).

  As shown in FIG. 4, the TG value (mg / dl) was measured only on the left side of the bar graph of FIG. 4 in the pig liver enzyme digested product administration group only at 4 weeks (24 weeks of age in FIG. 4). A significant difference was observed between the non-administered group (the bar on the right side of the bar graph in FIG. 4). On the other hand, the leptin level (pg / ml) continued to differ after the 4th week of ingestion.

  Regarding the proportion (%) of the organ weight at the time of dissection, as shown in FIG. 5, the swine liver enzyme degradation product administration group (the left bar in the bar graph in FIG. 5) is the non-administration group (the right bar in the bar graph in FIG. 5). ) And the fat weight was significantly lower.

  Regarding the blood component analysis at the time of dissection, as shown in FIG. 6, lipid peroxide (nmol / ml), creatine (mg / ml), uric acid (mg / ml), albumin (g / dl), GOT (Karmen unit) ), GPT (Karmen unit) was measured, but the pig liver enzyme hydrolyzate administration group showed a significantly low value in lipid peroxide. There was no significant difference in other items.

  For fecal analysis at the 14th week of test food intake (at the time of dissection at 32 weeks of age), dry fecal mass (g / day), fecal fat mass (mg / day), fat digestibility (%), neutral sterols (Μmol / l) and acidic sterol (μmol / l) were measured, and the swine liver enzyme degradation product administration group showed significantly high values for dry feces, fecal fat, fat digestibility, and neutral sterols. On the other hand, there was no significant difference in acidic sterols.

The above is summarized as follows.
(1) The body weight, blood glucose level, blood cholesterol, and blood leptin were significantly reduced by long-term intake of the porcine liver enzyme degradation product.
(2) Based on organ weight, accumulation of perirenal fat was suppressed by ingestion of porcine liver enzyme degradation product.
(3) From the analysis of blood components, a decrease in lipid peroxide was observed due to intake of porcine liver enzyme digest.
(4) From fecal analysis, it was confirmed that intake of porcine liver enzyme degradation product promotes fat excretion (neutral sterol) in feces.

  From the above test examples, it was demonstrated that the porcine liver enzyme degradation product has an effect of contributing to improvement of obesity and lipid metabolism. Therefore, the present invention is based on such a demonstration, to test meal A containing an enzymatic degradation mixture having an amino acid as a main component and having a molecular weight of 50 to 1000, obtained by concentrating an enzyme-treated component of animal liver. The corresponding food has an effect of improving lipid metabolism and sugar metabolism.

(Test Example 2)
In Test Example 1, it was demonstrated that the porcine liver enzyme degradation product has an effect of contributing to improvement of obesity and lipid metabolism. The two types of test meals A and B used in Test Example 1 are common in that they contain liver powder as described above, but have different fractional molecular weights.

  By the way, in order to compare the effects of the above-mentioned two types of test meal A and test meal B, as Test Example 2, an administration test of test meal A and test meal B was further performed. As a result of this administration test, it was found that test meal A (fraction molecular weight 50 to 1000) was superior in lipid metabolism improvement effect and sugar metabolism improvement effect compared to test meal B (fraction molecular weight 1000 to 100,000). . Hereinafter, the administration test of Test Example 2 will be described using Test Food A as a test food according to the present invention and Test Food B as a comparative example.

  For the administration test, type 2 diabetes model rat OLETF (Otsuka Long-Evans Tokyo Fatty) male 18-week-old (n = 20) was prepared, and oral glucose tolerance test (2.0 g glucose / kg b.w) at 19 weeks. )), And based on the results, the groups were divided so that there was no bias (five animals in each group).

  Then, ingestion of test meal A (containing 20% of liver powder with a fractional molecular weight of 50 to 1000) was started from the 20th week, and blood was collected at fasting every 4 weeks thereafter to determine the body weight and cholesterol of the blood that was determined. Free fatty acids, neutral fats, blood glucose levels, plasma insulin levels, and leptin concentrations were measured by standard methods.

  On the other hand, the same administration test as the test meal A according to the present invention was performed for the comparative example (test meal B: containing 20% of liver powder having a fractional molecular weight of 1000 to 100,000).

  As a result of these administration tests, the lipid metabolism improvement effect and the sugar metabolism improvement effect were evaluated by 10-step evaluation, and these are shown in Table 2. In addition, 10 shows the highest and 1 shows the lowest effect.

  As a result, it was demonstrated that the test meal A according to the present invention (containing 20% of liver powder having a molecular weight of 50 to 1000) has significantly higher lipid metabolism improvement effect and sugar metabolism improvement effect than the comparative examples. .

  Next, Examples 1-3 of the food containing the enzymatic degradation mixture of animal liver according to the present invention will be given and the effects thereof will be described below. Example 1 was prepared by adding 20% of liver powder having a molecular weight of 50 to 1000 to a beverage in the same manner as in the test food A according to the present invention (hereinafter referred to as “beverage containing 20% pork liver powder”). ").

  In Example 2, 20% liver powder having a fractional molecular weight of 50 to 1000 and 2% chitosan were added to a beverage to make a food (beverage containing 20% pork liver powder + 2% chitosan).

  Furthermore, in Example 3, 20% liver powder having a fractional molecular weight of 50 to 1000 and 2% containing beverage indigestible dextrin were added to the beverage to make a food (porcine liver powder 20% + containing beverage difficult It is called “a beverage containing 2% digestible dextrin”).

  In order to demonstrate the effects of these Examples 1 to 3, an intake experiment was performed in comparison with Comparative Examples 1 to 3 described below.

  Comparative Example 1 is a beverage containing 2% chitosan that does not contain liver powder. Comparative Example 2 is a beverage containing 2% indigestible dextrin containing beverage that does not contain liver powder. It is an additive-free beverage that does not contain any of pork liver powder, chitosan, and contained beverage indigestible dextrin.

  In the ingestion experiments of Examples 1 to 3 and Comparative Examples 1 to 3, 40 adult volunteers with a BMI of 25 or more were divided into 10 test groups for each 10 so that the body fat percentage was not biased. Ingested 3 times a day in the morning and night every day for 8 weeks, the degree of decrease in body fat after 8 weeks was taken as 100% body fat before feeding, and the body fat reduction rate was measured to reduce body fat The effect was confirmed.

  The results of the ingestion experiment are shown in Table 3 below. According to Table 3, it was demonstrated that Examples 1 to 3 were more effective in reducing body fat than the comparative examples. In particular, as in Example 2 and Example 3, it was proved that the effect was further enhanced by containing chitosan and a beverage resistant digestible dextrin.

(Test Example 3)
In order to clarify the functional body of the porcine liver enzyme degradation product, the present inventors have further conducted experiments and studies on lipid accumulation of porcine liver enzyme degradation product (hereinafter also referred to as “PLH”) using visceral adipocytes. Went. The experimental method is as follows.

1. Test method (1) As a sample, PLH prepared by the method described above was used.

(2) Test process using mesenteric adipocytes, examination a. Mesenteric adipocyte culture As the cells, mesenteric proadipocytes (manufactured by Primary Cell Co., Ltd.) collected from the mesentery of Wistar male rats were used. That is, mesenteric adipocytes prepared to be 1 × 10 ⁵ cells / ml / well were seeded in a 24-well plate and cultured overnight in a CO ₂ incubator (37 ° C., 5% CO ₂ ).

B. Preparation of Porcine Liver Enzymatic Degradation Product-Added Medium Porcine liver enzymatic degradation product (PLH) was dissolved in the medium so as to have respective concentrations of 0.01, 0.04, 0.1 and 1 mg / ml.

C. Mesenteric adipocyte culture with porcine liver enzyme-decomposed product-added medium The medium was changed after overnight culture, and a prepared porcine liver enzyme-degraded product (PLH) -added medium was added and cultured in a CO ₂ incubator for 3 days. Meanwhile, on the second day of culture, the medium was changed, and a newly prepared medium containing PLH was added.

D. Collection of mesenteric adipocytes and preparation of measurement sample After completion of the culture, the medium was removed and washed with PBS. Cells were collected with the enzyme extract, disrupted with an ultrasonic disrupter, and centrifuged at 12,800 × g and 4 ° C. for 5 minutes. The supernatant was recovered and used as a measurement sample.

E. Measurement of intracellular neutral fat The measurement was performed using a commercially available kit (Triglyceride E-Test Wako).

(3) Statistical processing Data of both groups obtained from the experiment are shown as mean ± standard error. Tested by repeated measure one-way ANOVA, and when the F value was significant, multiple comparison was performed using Fisher's PLSD method.

2. Test results (1) Influence on lipid accumulation FIG. 8 shows the measurement results of intracellular triglycerides. According to FIG. 8, in the medium supplemented with 0.1 and 1 mg / ml PLH, a significant decrease in intracellular neutral fat was observed as compared with the medium without additive. From this, it was newly clarified that PLH has an action of inhibiting fat accumulation on mesenteric fat cells which are visceral fat.

  Many evaluations using adipocytes have already been reported with 3T3-L1 cells, which are established mouse fibroblasts. Among these, as food ingredients, tea catechin fat accumulation inhibition and lipolysis effects, DHA (docosahexaenoic acid) fat accumulation inhibition effects, and adipocyte differentiation inhibition effect by grape seed extract have been reported as papers.

  Recently, when a thiazolidine derivative, which is a therapeutic agent for diabetes, is added to a culture medium using a primary culture system of mesenteric adipocytes, which is representative of visceral fat, and 3T3-L1 cells, there are reports of different reactivity. ing. Therefore, the present inventors pay attention to this method, and consider that the evaluation by the primary culture system close to the in vivo condition is also suitable for the evaluation using cells, and in this test, the evaluation is performed using mesenteric fat cells. Went.

  In this test, the method of adding the porcine liver enzyme degradation product directly to the medium was examined. Since the free fatty acid level in blood of OLETF rats tended to be low in the porcine liver enzyme degradation product (PLH) group, it was considered that the porcine liver enzyme degradation product does not affect the degradation of adipocytes per se. However, in visceral adipocytes most closely related to insulin resistance and metabolic syndrome among adipocytes, it was possible to obtain the knowledge that a porcine liver enzyme degradation product acts to suppress lipid accumulation.

(Test Example 4)
As a result of Test Example 3, it was confirmed that the digested porcine liver enzyme acts on the suppression of lipid accumulation in visceral fat cells. Next, in Test Example 4, measurement was performed using a visceral fat cell system. Based on the components in the porcine liver enzyme degradation product (PLH), the search for substances that contribute to the suppression of lipid accumulation (exploration of effect-related components) was advanced. If a substance that contributes to the suppression of lipid accumulation can be searched, when the substance is added to food, it is not particularly necessary to be derived from porcine liver, and may be derived from a synthetic and / or natural product corresponding to the substance.

  From this point of view, in Test Example 4, the component composition of the porcine liver enzyme degradation product (PLH) prepared in Test Example 3 was analyzed in advance. As a result, Table 4 shown in FIG. 9 and Table 5 shown in FIG. 10 were obtained. Based on the analysis results, a vitamin mixture, a mineral mixture, and an amino acid mixture were prepared from synthetic and / or natural products, respectively. In Test Example 4, these are used as specimens.

1. Test Method (1) Specimen As a specimen, the porcine liver liver enzyme degradation product (PLH) prepared in Test Example 3 is used. Furthermore, as described above, as described above, the numerical value (see Table 4 shown in FIG. 9 and Table 5 shown in FIG. 10) obtained by analyzing the porcine liver enzyme degradation product (PLH) prepared in Test Example 3 is shown below. (In FIG. 11: vitamin), a mineral mixture (in FIG. 11: mineral), and an amino acid mixture (in FIG. 11: amino acid) are prepared separately and used.

(2) Test process using visceral adipocytes, examination a. Culture of mesenteric adipocytes The mesenteric adipocytes were cultured in the same manner as in Test Example 3 above.

B. Preparation of supplemented medium A medium containing 1 mg / ml of porcine liver enzyme degradation product (PLH) as a specimen was prepared. Furthermore, three types (a vitamin mixture, a mineral mixture, and an amino acid mixture containing constituent amino acids) are prepared as a mixture according to the amount of nutrients contained in 1 mg / ml porcine liver enzyme degradation product (PLH), and each of these is contained. A medium was prepared. Thereafter, culture and measurement of neutral fat were carried out by the same method as in Test Example 3 above.

2. Test results (1) Influence on lipid accumulation FIG. 11 shows the measurement results of intracellular triglycerides. According to FIG. 11, a significant decrease in intracellular triglycerides was recognized by adding PLH and an amino acid mixture, respectively, compared to the medium without additive.

  Thus, it was newly clarified that the amino acid mixture constituting the porcine liver enzyme degradation product (PLH) has an action of inhibiting fat accumulation on mesenteric fat cells which are visceral fat. On the other hand, the vitamin mixture and mineral mixture constituting PLH showed no significant difference in lipid accumulation as compared with the case of no addition, and the vitamin mixture tended to promote lipid accumulation rather.

  So far, there has been no report on the addition of nutrients using mesenteric adipocytes, but in the established cell line 3T3-L1, the addition of vitamin B6 and vitamin C enhances the accumulation of neutral fat. Although there is a difference in properties between primary cultured cells and cell lines, a direct comparison is not possible, but the vitamin mixture may act to suppress lipid accumulation in mesenteric adipocytes Is considered low.

  From the viewpoint of obesity prevention, it was very meaningful that the suppression of lipid accumulation was confirmed in Test Examples 3 and 4, and an amino acid mixture was cited as a component involved, and an effective component could be identified. From these results, it was clarified that the lipid accumulation was significantly reduced by the addition of the amino acid mixture, and the contribution of amino acids and peptides as protein sources could be verified.

(Test Example 5)
As a result of the test example 4, in visceral fat cells, it was confirmed that the lipid accumulation was significantly reduced by addition of the amino acid mixture and that the amino acid group had activity. Using the system, a search was made as to which amino acid group had an effect (search for an amino acid group related to the effect).

1. Test method (1) Specimen Here, amino acids were divided into several groups for experiments. As a classification method for this classification, casein, which is usually used as a standard control for protein in experimental animal food, is included. Of the amino acid composition of the casein diet (test food A of Test Example 1) and the amino acid composition of the pig liver enzyme-decomposed processed food (test food C of Test Example 1) containing porcine liver enzyme-decomposed processed food as protein Below (Table 6 shown in FIG. 12), groups G1 to G5 of the following specimens were respectively prepared.

Group G1: Amino acid group (glycine + alanine) more than casein diet
G2 group: Amino acid group (cysteine, asparagine) slightly higher than casein diet
Group G3: branched chain amino acid group <BCAA> (Val + Leu + Ile)
G4 group: fewer amino acids than casein diet (Thr + Ser + Met + Phe + His + Lys + Trp)
G5 group: Less amino acid group than casein diet (Glu + Pro + Tyr + Arg)

(2) Test process using visceral adipocytes, examination a. Culture of mesenteric adipocytes The mesenteric adipocytes were cultured in the same manner as in Test Example 3 above.

B. Preparation of supplemented medium Five mixtures (G1 group: glycine + alanine), (G2 group: cysteine + asparagine), (G3 group: each group 5) of G1 to G5 according to the amount of amino acids contained in PLH Val + Leu + Ile), (G4 group: Thr + Ser + Met + Phe + His + Lys + Trp), and (G5 group: Glu + Pro + Tyr + Arg). Thereafter, culture and measurement of neutral fat were carried out in the same manner as described in Test Example 3 above.

2. Test results (1) Influence on lipid accumulation FIG. 13 shows the measurement results of intracellular triglycerides. According to FIG. 13, a significant decrease in intracellular triglycerides was recognized by adding a mixture of glycine and alanine in the G1 group as compared with a non-added medium (indicated by 0 in FIG. 13). From the above, it was clarified in Test Example 5 that a mixture of glycine and alanine, which is a structurally simple low molecular weight neutral amino acid, has activity.

  In addition, as a function of glycine, it has been reported that glutathione and a raw material of porphyrin that is a hemoglobin component are used. On the other hand, as a function of alanine, it is one of the amino acids most easily used as an energy source, Although it has been known that it has an action to improve alcohol metabolism and can be used as a material for synthesizing sugar necessary for the body, it is the present invention that a mixture of glycine and alanine causes a decrease in intracellular neutral fat. This became clear for the first time in Test Example 5.

(Test Example 6)
As a result of Test Example 5 above, it was confirmed that a mixture of glycine and alanine, which is an amino acid component rich in porcine liver enzymatic degradation product, was active against visceral adipocytes. In 6, the visceral fat cell system was used to search for glycine alone, alanine alone, and the synergistic effect of glycine and alanine (search for effect-related amino acids).

1. Test Method (1) Specimens In order to examine the synergistic effect exhibited by glycine and alanine, six groups of I to VI were tested as specimens, with each other's content ratio (see the column of Gly: Ala in Table 7 below). Each group was created.

(2) Test process using visceral adipocytes, examination a. Culture of mesenteric adipocytes The mesenteric adipocytes were cultured in the same manner as in Test Example 3 above.

B. Preparation of supplemented medium Five types of glycine and alanine mixtures (groups I to VI) described in Table 7 were prepared according to the amino acid amounts of glycine and alanine containing PLH. In addition, an additive-free group (VI) was also prepared as a control group. Thereafter, culture and measurement of neutral fat were carried out in the same manner as in Test Example 3 above.

2. Results (1) Influence on lipid accumulation Table 7 shows the measurement results of intracellular triglycerides. According to this Table 7, it is preferable that glycine and alanine, which are structurally simple low molecular weight neutral amino acids, each have activity alone or in a mixture mixed in any ratio, preferably at a certain ratio (glycine: alanine = 1). : 9 to 9: 1) has an activity, more preferably a certain ratio (glycine: alanine = 1: 9 to 5: 5) has an activity.

  In Example 4 of the present invention, pig liver is crushed, reacted with protease at 45 ° C. at an optimum pH, extracted under pressure, concentrated by centrifugation, and the molecular weight is reduced to 50 to 1000 lever powder by ultrafiltration. A food containing an amino acid mixture derived from a synthetic and / or natural product having an amino acid composition equivalent to that of an enzymatic degradation mixture of an animal liver that has been adjusted.

  This Example 4 was conceived based on the findings of Test Example 4 above based on the knowledge that a synthetic and / or natural product-derived amino acid mixture having an amino acid composition equivalent to that of an enzymatic degradation mixture of animal liver has activity. That is, from the results of Test Example 4, as long as it is a food containing an amino acid mixture derived from an equivalent synthetic and / or natural product, it is not an enzymatic degradation mixture of animal liver as in Examples 1 to 3. It is based on what is good.

  Example 5 of the present invention is a synthetic and / or natural product derived amino acid glycine alone, or alanine alone, or a certain ratio of glycine and alanine (glycine: alanine = 1: 9-9: 1). A food containing a mixture.

  Combine meat ingredients (pork; 70.2 parts per 100 parts by weight of the kneaded raw material mixture) and pork back fat (4.3 parts) and use a meat grinder or silent cutter to grind it into a meat lump with a diameter of about 15 mm or less. , Ice (15.5 parts), salt (1 part), vitamin C (sodium ascorbate, 0.1 part), polymerized phosphate (0.2 parts), sugar and spices (0.7 parts) ), Potato starch (3 parts) and glycine alone, or alanine alone, or a mixture of glycine and alanine in a certain ratio (glycine: alanine = 1: 9-9: 1) added to each group (5 parts) Then, using a silent cutter or a vacuum mixer, prepared meat for sausage was prepared. This was then filled into a sheep intestine or collagen casing, twisted and ligated with a length of 5 to 10 cm, suspended on a suspension rod, dried, cooked until the center temperature reached 72 ° C., and cooled.

  A human test was performed to evaluate the wiener sausage of the present invention obtained above. In other words, 5 men and women in each group have 2 meals (about 10 g / tube) every meal (3 meals a day) for 2 weeks, and blood is collected before and after taking this sausage. The effect of the triglyceride in blood was measured.

  In Example 5, as shown in Table 8, the results of the same tendency as in Test Example 6 were obtained, and the structurally simple amino acid glycine alone, or alanine alone, or glycine and alanine were present. This is based on the finding that a mixture of a certain ratio (glycine: alanine = 1: 9 to 9: 1) has an activity of significantly reducing intracellular triglycerides.

  As mentioned above, although the best form for implementing the food containing the enzyme degradation mixture of animal liver concerning the present invention was explained based on a test example, an example, etc., the present invention is such a test example, an example. It goes without saying that there are various embodiments within the scope of the technical matters described in the claims.

  As described above, pork liver powder containing an enzymatic degradation mixture of animal liver of the present invention and / or an amino acid mixture having an equivalent amino acid composition and / or glycine alone and / or alanine alone and / or a mixture of glycine and alanine are used daily. It can be applied as a food for improving lifestyle-related diseases by containing it in meat products such as staple foods, ham, sausage, hamburger, meat dumplings, tablet-like supplements or beverages.

It is a figure which shows the change of the body weight during the test period by the test example of this invention. It is a graph which shows the change of the blood glucose level by the test meal intake by the test example of this invention, and an insulin level. It is a graph which shows the change of the cholesterol value by the test example of this invention, and a NEFA value. It is a graph which shows the change of TG and leptin by the test example of this invention. It is a graph which shows the change of the organ weight at the time of dissection by the test example of this invention. It is a graph which shows the result of the blood component analysis at the time of dissection by the test example of this invention. It is a graph which shows the result of feces analysis in the test intake 14 weeks by the test example of this invention. It is a graph which shows the effect of a porcine liver enzyme degradation product (PLH) in the fat accumulation of the mesentery fat cell by the test example of this invention. It is a table | surface which shows the component composition of a pig liver enzyme decomposition product (PLH) in the test example of this invention. It is a table | surface which shows the amino acid composition of a porcine liver enzyme degradation product (PLH) in the test example of this invention. It is a graph which shows the effect of a pig liver protein hydrolyzate (PLH) in the fat accumulation of the mesentery fat cell by the test example of this invention. It is a table | surface which shows the amino acid composition of the casein diet and the pig liver protein hydrolyzate diet in the test example of this invention. It is a graph which shows the influence on the fall of the intracellular triglyceride by the glycine and alanine mixture addition by the test example of this invention.

Claims (6)

  An enzyme-degraded mixture of animal liver obtained by concentrating components obtained by enzymatically treating animal liver and improving lipid metabolism and sugar metabolism.   2. The enzymatic degradation mixture of animal liver according to claim 1, wherein the enzymatic degradation mixture is an enzymatic degradation mixture having a molecular weight of 50 to 1000 containing amino acids, water-soluble vitamins, minerals, nucleic acids and peptides.   The enzyme degradation mixture is prepared by pulverizing pork liver, reacting with protease at 45 ° C. at an optimum pH, extracting with pressure, concentrating, and adjusting the molecular weight to 50 to 1000 liver powder by ultrafiltration. 2. The enzymatic degradation mixture of animal liver according to claim 1.   Equivalent to an enzymatic degradation mixture of animal liver in which pork liver is crushed, reacted with protease at 45 ° C at an optimal pH, extracted under pressure, concentrated, and molecular weight adjusted to 50-1000 liver powder by ultrafiltration An enzymatic degradation mixture comprising a synthetic and / or natural product-derived amino acid mixture having the amino acid composition of:   An enzymatic degradation mixture comprising glycine or alanine or a mixture of glycine and alanine, characterized by improving lipid metabolism and sugar metabolism.   The foodstuff containing the enzyme decomposition mixture in any one of Claims 1-5.

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