JPH0545220B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Application Field] The present invention relates to a protein material suitable as a protein source for, for example, food, feed, fodder, pet food, etc., and particularly relates to partially decomposed fish protein that has not been heat denatured, that is, partially decomposed fish protein. The present invention relates to a protein material consisting of. [Prior art and problems to be solved by the invention] Conventionally, as protein sources for foods, feeds, fodder, pet foods, etc., vegetable protein raw materials such as soybeans, peanuts, cottonseeds, sesame, sunflowers, and wheat, and their Vegetable protein sources such as defatted processed products and substances derived therefrom, and animal sources such as various milks, livestock meat, fish meat, other animal meats, eggs and their processed products, and substances derived therefrom. Protein sources are used, and more versatile protein materials include isolated proteins separated from the protein raw materials mentioned above, such as vegetable proteins such as soybean protein and wheat protein, and casein and fruit protein concentrate ( Fish Protein
Concentrates (hereinafter referred to as FPC),
Animal proteins such as meal are also used. Among these versatile protein materials, vegetable proteins are cheap and most commonly used, but their amino acid compositions, especially essential amino acids, include lysine,
It has a low content of methionine, etc., and its nutritional value is inferior to that of animal protein, which contains higher amounts of methionine. Therefore, for example, animal protein such as fish meal is used as an excellent protein source in animal feed, and fish meal and FPC for food use are already available. However, these fish protein materials for food and feed are heat-treated during the manufacturing process,
Due to the use of solvents, the protein is heat denatured, making it difficult to digest, and the epsilon amino group of lysine, which has a free epsilon amino group, which is said to be effective as an essential amino acid, may be difficult to digest with other active substances. It is not always satisfactory because it binds and becomes less susceptible to decomposition by digestive enzymes, and trace amounts of solvent may remain in the product. [Means for Solving the Problems] In view of the above-mentioned circumstances, the inventors of the present invention have conducted intensive studies to develop a fish protein material that does not have the drawbacks of conventional fish meal, FPC, etc. We have arrived at the present invention. That is, the present invention provides fish proteins that are partially decomposed and not heat denatured, with less than 20% having a molecular weight of more than 100,000, and less than 20% having a molecular weight of 100,000 to 40,000.
20-50%, 20-50% with a molecular weight of 40,000-14,000;
This is a protein material made of partially decomposed fish protein, in which less than 20% of the protein has a molecular weight of less than 14,000. The protein material of the present invention will be explained in detail below. The above-mentioned partially decomposed fish protein used in the present invention is, for example, (1) a fish body as it is or one from which internal organs and/or epidermis have been removed, (2) fish meat obtained by collecting meat from a fish body, or fish meat obtained by collecting meat from a fish body, or Processed materials and (3) raw materials selected from fish residues, etc. are treated with proteolytic enzymes or autolyzed, and then, if necessary, fish bones and fish oil, or further water-soluble components are separated and removed. obtained by. Specific examples of the raw materials include whole bodies of highly catchy fish such as herring, sardine, mackerel, saury, Japanese sardine, walleye pollock, flounder, sardine, and pilchard; internal organs and/or the like;
and fish meat from which the skin has been removed; fish meat such as fallen meat or frozen fallen meat obtained by harvesting them; processed fish meat products such as dehydrated meat and surimi obtained by processing them; These include residues of walleye pollock discharged from canning factories, and residues of fish such as bonito, tuna, salmon, trout, and mackerel discharged from canning factories. In order to obtain protein, it is preferable to use the whole body of a highly fresh fish that is often caught.If the purpose is to obtain protein material for food, the head, internal organs, and/or epidermis, etc. should be removed from the whole fish. It is preferable to use fish meat and its processed products. Examples of proteolytic enzymes used to obtain the above partially degraded fish protein include acrosin,
Urokinase, uropepsin, elastase, enteropeptidase, cathepsin, kallikrein, kininase 2, chymotrypsin, chymopapain, collagenase, streptokinase, subtilisin, thermolysin, trypsin, thrombin, papain, pancreatopeptidase, huicin, plasmin, renin, reptilase, rennin, etc. proteases such as; aminopeptidases such as arginine aminopeptidase, oxytocinase, leucine aminopeptidase;
Angiotensinase, angiotensin converting enzyme, insulinase, carboxypeptidase such as arginine carboxypeptidase, kininase 1, thyroid peptidase, dipeptidase such as carnosinase, prolinase, peptidase such as pronase; and other proteolytic enzymes and their They include modified products, blended products, etc., and are divided into exo-type proteases that act from the end of the polypeptide chain and endo-type proteases that act internally, depending on their mode of action, with endo-type proteases being particularly preferred. In order to obtain the partially decomposed fish protein used in the present invention, the above-mentioned raw materials are first treated with the above-mentioned proteolytic enzymes or autolyzed, but when treated with the proteolytic enzymes, the degree of treatment is Increase rate of soluble nitrogen after enzyme treatment relative to total nitrogen in the raw material: Ne−No/Nt×100 (where, in the formula, Nt is the weight% of the total nitrogen in the raw material,
Ne is the weight percent of soluble nitrogen in the product after enzyme treatment, and No is the weight percent of soluble nitrogen in the product after treatment under the same conditions except that no enzyme is added).
The treatment may be carried out until the concentration is reduced to 50%, preferably 5 to 40%.
C., preferably 30 to 60.degree. C., for about 5 minutes to 2 hours, preferably 10 minutes to 1 hour, with stirring. Further, the amount of enzyme used in this case is usually 0.005 to 1.0% by weight based on the raw material to be treated. In addition, when the above-mentioned raw material is subjected to self-digestion, the increase rate of soluble nitrogen after self-digestion with respect to the total nitrogen in the starting raw material: Ne−No/Nt×100 (However, in the formula, Nt is the total nitrogen in the raw material. weight%,
Ne is the weight percent of soluble nitrogen in the product after autolysis, No is the weight percent of soluble nitrogen in the raw material)
Autolysis may be carried out so that the amount of oxidation is 10 to 50%, preferably 10 to 40%, for example, at 30 to 60°C, preferably 40 to 60°C, for about 20 minutes to 2 hours, preferably 30 minutes. Self-extinguishing may be performed while stirring for ~1 hour. The product obtained by treatment with a proteolytic enzyme or the product obtained by autolysis as described above is
After deactivating the enzyme by various means, if the raw material used contains large amounts of fish bones, fish oil, etc., these can be removed from the product by centrifugal filtration, centrifugation, etc. The partially degraded fish protein used in the present invention can be obtained by removing the aqueous solution portion containing water-soluble components from the product by, for example, centrifugation. The above separation is particularly preferable if a two-layer separator or a three-layer separator is used because continuous treatment is possible. The partially decomposed fish protein used in the present invention has a molecular weight of 100,000 or more, with a molecular weight of 100,000 to 20%.
20 to 50% have a molecular weight of 40,000 to 14,000, 20 to 50% have a molecular weight of 40,000 to 14,000, and 20% have a molecular weight of less than 14,000.
% or less is essential. The ratio of each molecular weight category is determined by the method of Kondo et al. (Biochemistry, Vol. 44,
p. 304, 1972), sodium phosphate/
6 μg of the sample was injected into an SDS/polyacrylamide gel using SDS (PH7.2), and electrophoresis was performed at 40 mA for 7 hours.
The electrophoresis pattern was recorded using
After creating a calibration curve using this, the molecular weight of the protein in the sample is determined from the calibration curve, and using a scanning densitometer, the molecular weight is determined to be 100,000 or more, 100,000 to 40,000, 100,000 to 40,000,
The ratio of proteins in four categories, 40,000 to 14,000 and less than 14,000, was measured, and the protein was treated with the proteolytic enzyme or self-treated so that the ratio of each molecular weight category fell within the above range. The degree of digestion and the rate of separation of the aqueous portion containing water-soluble components from the product after treatment are selected. The partially decomposed fish protein used in the present invention can be obtained as described above, but in particular, it can be obtained from the fish body as it is.
Alternatively, partially decomposed fish protein obtained by removing the internal organs and/or epidermis, treating with a proteolytic enzyme, and then separating and removing the fish bone, fish oil, and aqueous solution is preferable. Incidentally, such partially decomposed fish protein can also be dried by various means such as freeze drying, spray drying, ventilation drying, etc., if necessary. [Example] Example 1 To 1 kg of sardines, 0.3 g of a proteolytic enzyme: Protease Amano A (manufactured by Amano Pharmaceutical Co., Ltd.) was dissolved in a small amount of water, and stirred for 30 minutes while maintaining the temperature at 50°C. The fish meat separates from the fish bones and the whole becomes a slurry. The rate of increase in soluble nitrogen in this slurry was measured by the following method. Take 10g of slurry and add 30g of water
ml, add 5 ml of 10% trichloroacetic acid solution, make up to 50 ml with water, and filter through filter paper (Toyo Roshi: No. 5A). After decomposing 10ml of this filtrate with sulfuric acid using a conventional method,
Measure the amount of soluble nitrogen using the Kjeldahl method and define it as soluble nitrogen (Ne). In addition, 2 g of slurry was taken and treated in the same manner after decomposition with sulfuric acid to convert it to total nitrogen (Nt). Furthermore, after keeping the enzyme-free raw material at 50°C for 30 minutes, take 10g of the raw material, add trichloroacetic acid solution in the same way, and filter the filtrate (10ml) with sulfuric acid to measure the amount of soluble nitrogen, which is referred to as soluble nitrogen (No). As a result of the above measurement, the rate of increase in soluble nitrogen in this slurry: Ne-No/Nt×100 was 24.2%. Next, this slurry was heated and kept at 75℃ for 15 minutes to inactivate the enzyme, and then the fish bones were removed using a basket-type centrifuge equipped with a 6-mesh stainless steel wire mesh. The slurry was heated to 3000r.
The mixture was centrifuged at pm for 5 minutes to separate fish oil, water phase, and partially decomposed fish protein precipitate to obtain a cake consisting of partially decomposed fish proteins. This cake was vacuum freeze-dried to obtain 130 g of powdered partially decomposed fish protein. Next, a small amount of this dried product was taken, thoroughly washed with cold ethanol, dried under reduced pressure to remove the solvent, and the molecular weight of the protein was measured by electrophoresis using SDS/polyacrylamide gel as follows. . Kondo et al.'s method (Biochemistry, Vol. 44, p. 304,
Sodium phosphate/SDS (PH
Inject 6 μg of the sample into the SDS/polyacrylamide gel in 7.2), perform electrophoresis at 40 mA for 7 hours,
Similarly, an electrophoresis pattern is recorded using a standard molecular weight kit (manufactured by Pharmacia Japan), and a calibration curve is prepared using this. Determine the protein molecular weight in the sample from a calibration curve, and use a scanning densitometer to determine whether the molecular weight is over 100,000, 100,000 to 40,000, or 40,000 to 40,000.
The proportion of proteins between 10,000 and 14,000 and less than 14,000 was measured. As a result, the above partially degraded fish protein has a molecular weight of 10
7% are over 10,000, 41% are between 100,000 and 40,000, 4
35% is between 10,000 and 14,000, 17% is less than 14,000.
It was hot. Similar tests were conducted on starting materials, and the results showed that 34% had a molecular weight of over 100,000, and 28% had a molecular weight of 100,000 to 40,000.
%, 30% is between 40,000 and 14,000, and 8% is less than 14,000. It can be seen that it is molecularized. Example 2 When 1 kg of sardines is kept at a temperature of 50°C and stirred for 40 minutes, the fish meat gradually separates from the fish bones and the whole becomes a slurry. The rate of increase in soluble nitrogen in this slurry was measured by the following method. Take 10g of slurry and add 30g of water
ml, add 5 ml of 10% trichloroacetic acid solution, make up to 50 ml with water, and filter through filter paper (Toyo Roshi: No. 5A). After decomposing 10ml of this filtrate with sulfuric acid using a conventional method,
Measure the amount of soluble nitrogen using the Kjeldahl method and define it as soluble nitrogen (Ne). In addition, 2 g of slurry was taken and treated in the same manner after decomposition with sulfuric acid to convert it to total nitrogen (Nt). Further, 10 g of the raw material was taken, and 10 ml of the filtrate was filtered after adding a trichloroacetic acid solution in the same manner, and the filtrate was decomposed with sulfuric acid to measure the amount of soluble nitrogen, which was defined as soluble nitrogen (No). As a result of the above measurement, the rate of increase in soluble nitrogen in this slurry: Ne-No/Nt×100 was 16.0%. Next, this slurry was heated and kept at 75℃ for 15 minutes to lose enzyme activity, and then the fish bones were removed using a basket-type centrifuge equipped with a 6-mesh stainless steel wire mesh. slurry
The mixture was centrifuged at 3000 rpm for 5 minutes to separate fish oil, water phase, and partially decomposed fish protein precipitate to obtain a cake consisting of partially decomposed fish proteins. This cake was vacuum freeze-dried to obtain 115 g of powdered partially decomposed fish protein. The protein molecular weight distribution of this partially degraded fish protein is
11% are over 100,000, 46% are between 100,000 and 40,000,
33% are between 40,000 and 14,000, and 10 are less than 14,000.
It was %. Use Examples 1 and 2 and Comparative Use Example 1 Using the partially decomposed fish proteins obtained in Examples 1 and 2, feed raw materials were mixed and ground according to the formulation shown in the table below to prepare fish feed. For comparison, fish feed was prepared in the same manner using white fish meal.

[Table] 50 parts of water was added to 100 parts of the above feed, mixed thoroughly, and then granulated using a meat hopper.The pellets were fed to carp weighing approximately 150 g, and a breeding test was conducted for one month. The number of fish tested was 20 in each area, and the water tank was made of vinyl chloride. The results of the above test are shown in the table below.

【table】

〔Effect of the invention〕

The protein material of the present invention has a high content of methionine, lysine, etc. in its amino acid composition, particularly in its essential amino acid composition, and has superior nutritional value compared to vegetable proteins that have a low content of methionine and lysine. Because it is decomposed, it is easily digested by humans and animals, and it is also used in the heat treatment process and FPC during fishmeal production.
Since the manufacturing process does not include a solvent extraction process, there are no problems such as heat denaturation of the protein or residual solvent, making it an extremely excellent protein material. The protein material of the present invention can be widely used as a substitute for soybean protein, fishmeal, FPC, etc., or as a new source of animal protein, for example, in foods, feeds, fodder, pet foods, and the like.

Claims (1)

[Scope of Claims] 1 Partially decomposed, undenatured fish protein with a molecular weight of more than 100,000 20%
The following is a protein material made of partially decomposed fish protein in which 20-50% has a molecular weight of 100,000 to 40,000, 20-50% has a molecular weight of 40,000 to 14,000, and 20% or less has a molecular weight of less than 14,000. . 2 Partially decomposed fish protein can be used in the fish body as it is, or after removing the internal organs and/or the epidermal part,
treated with proteolytic enzymes or autolyzed;
The protein material according to claim 1, which is a partially decomposed fish protein obtained by subsequently separating and removing fish bones and fish oil, or water-soluble components. 3. A patent characterized in that the partially decomposed fish protein is a partially decomposed fish protein obtained by treating fish meat with a proteolytic enzyme or autolyzing it, or by further separating and removing fish oil or fish oil and water-soluble components. The protein material according to claim 1. 4 Increase rate of soluble nitrogen after enzyme treatment with respect to total nitrogen in the raw material: Ne−No/Nt×100 (where, in the formula, Nt is the weight% of the total nitrogen in the raw material,
Ne is the weight percent of soluble nitrogen in the product after enzyme treatment, and No is the weight percent of soluble nitrogen in the product after treatment under the same conditions except that no enzyme is added).
The protein material according to claim 2 or 3, characterized in that the protein material is treated until it reaches 50%. 5 Increase rate of soluble nitrogen after autolysis relative to total nitrogen in the raw material: Ne−No/Nt×100 (where, in the formula, Nt is the weight% of the total nitrogen in the raw material,
Ne is the weight percent of soluble nitrogen in the product after autolysis, No is the weight percent of soluble nitrogen in the raw material)
3. The protein material according to claim 2 or 3, wherein the protein material is treated until the amount of the protein becomes 10 to 50%.