JPS6163247A - Production of globin protein - Google Patents

Abstract

PURPOSE:To obtain the titled protein easily and inexpensively, by collecting hemoglobin from animal blood, adding water and an inorganic or organic acid to dissociate heme and globin, adding a polyacrylic acid to the solution, agglomerating and removing heme. CONSTITUTION:Blood of domestic animal such as bovine, swine, sheep, etc. is hygienically collected, and hemoglobin is separated. The prepared hemoglobin is dissolved in water to give an aqueous solution, further hemolyzed by stirring, etc., blended with an inorganic or organic acid, and the aqueous solution is adjusted to <=3.0pH. Consequently, heme and globin of hemoglobin lose bond strength and are dissociated. Then, a polyacrylic acid (salt) is added to the adjusted hemoglobin solution, heme is agglomerated by stirring and removed, to give the aimed protein.

Description

[Detailed description of the invention]

Industrial Application Field The present invention is directed to the production and production of globin protein from animal blood.
It concerns how to Conventional technology In F grains of livestock and poultry such as IJ (cattle, pigs, sheep, chickens), a large amount of blood is obtained when slaughtered. This animal blood can be used as plasma powder in food layers, animal feed, adhesives, binders such as Sorceno, and as a raw material for the pharmaceutical drug Ptovorfiline 2Ng. Production of hemin. Some of it is used for things like protein production, but most of it is thrown away and has no use for it. However, it is extremely difficult to dispose of it as waste because it contains organic substances such as proteins at a fairly high concentration. The proteins that are present in approximately 15% of animal blood are mainly fulbumin and guprin, which are present in the serum fraction and blood depressant fraction, and globin protein, which binds to hemoglobin-heme to form hemoglobin. Among these, serum proteins are the ones for which separation techniques have been established and are being used industrially. Since it is difficult to separate globin protein from heme and globin, it has not been separated from animal blood for use. There are several methods known in the literature for separating globin from heme, such as separating globin by extracting heme in acidic acetone, which has been carried out in the laboratory and is relatively advantageous. However, this method is difficult to implement industrially because it requires a large amount of 77 tons, which is expensive, and there are also many problems in terms of operational safety. Also,
JP-A No. 56-106599 discloses that insoluble carboxymethyl cellulose contains hemoglobin in acidic water? A method is described in which glopin is isolated by bringing 8 liquids into contact and causing heme adsorption, but the carboxymethyl cellulose used in large amounts in this method is
Since it is a special and expensive product, it seems impossible to commercialize it in terms of cost, and if carboxymethyl cellulose is used repeatedly, there is a risk of microbial contamination. A problem to be invented or solved! It is an object of the present invention to provide a method for producing globin proteins that does not have the drawbacks such as double globin proteins. The purpose of the present invention is to provide a method of utilizing hemoglobin from blood collected from animals. The obtained hemoglobin is made into an aqueous solution, and the I of the aqueous solution is
The method is characterized in that hemoglobin is dissociated into heme and globin by setting +14 to 3.0 or less, and then polyacrylic acid or a salt thereof is added to aggregate and remove heme. Hereinafter, the method for producing this globin protein will be explained in order of steps. First, moglobin is separated from fresh blood hygienically collected from slaughtered animals.There are no restrictions on the method used, and conventional hemoglobin separation methods such as filtering coagulated blood, Any method for centrifuging blood containing an anticoagulant can be adopted. The obtained hemoglobin fraction has a solid content of 1 to 20%.
After dissolving in water to a certain concentration and further performing hemolysis by stirring, etc., add an inorganic acid such as hydrochloric acid or sulfuric acid or an organic acid such as citric acid or tartaric acid to adjust the pH to 3.0 or less, preferably about 1. Adjust to .9 to 2.5. As a result, the heme of hemoglobin and globin lose their binding strength and dissociate. When an aqueous solution of polyacrylic acid or its salt is then added to the pH-adjusted hemoglobin solution and stirred, the dissociated heme reacts with the flocculant to coagulate and precipitate. In this case, the polyacrylic acid or its salt is about 0.1 to 10% by weight (preferably 1 to 10% by weight) based on the hemoglobin in the liquid to be treated.
When added in the form of a 0.05 to 2.0 weight 1% aqueous solution, the filterability of aggregates and the removal rate of heme are good. The precipitate formed from the flocculant and heme is removed by filtration or centrifugation, yielding a tan-colored globin solution. There are various ways to obtain solid globin from this globin solution, but the DH is reduced to 5.5 by adding an alkali such as hydric soda.
When the temperature is adjusted to ~7.0 (preferably when heated), globin precipitates at an isoelectric point, so the most advantageous method is to collect it by filtration or centrifugation. The obtained globin precipitate is washed with a small amount of water to remove salts, and then freeze-dried and powdered to obtain a pale yellow-brown globin powder. Alternatively, a dilute alkali may be added to the water-washed globin precipitate to adjust the pH to approximately 7.3 to 8.0 to solubilize the globin, and the resulting aqueous solution may be spray-dried. The globin powder obtained as described above usually has a total nitrogen content of 11-12% and a citrus protein content of 65-7%.
It is about 5%. Note that the globin solution from which heme has been removed through the aggregation process still has a slight blood odor, so if it is treated with activated carbon or acid clay to also decolorize it, higher quality globin without the blood odor can be obtained. According to the production method of the present invention, globin protein is produced by using polyacrylic acid or its salt with a small titre and simply combining it with the simple 1) 11 temperature conditioning and solid-liquid separation operations. The present invention is expected to make it possible for the first time to industrially produce globin protein using animal blood as a raw material, as it does not require expensive equipment, chemicals, or solvents. It is thought that it can also contribute to solving the problem of animal blood disposal. In addition, the globin protein obtained by the production method of the present invention can be obtained by the benzuzine reagent method or the alkali-denatured globin hemochrome method.

[Koyotera is a high-quality product with no detectable heme and almost no blood odor, so it can be used as a high-quality, high-protein food ingredient, as an auxiliary raw material for ham, sausage, kamaboko, etc., or as an emulsifying agent. In addition to this, it can also be used to fortify the protein of pond foods, food for sick people, and so-called health foods, making it extremely useful. Example 1 Fresh clotted pig blood (hemoglobin content 10.7%)5
00g was centrifuged at 9000 rpm for 10 minutes to obtain hemoglobin as a blood clot. The clot was washed by dispersing it in 250+ sl of 0.9% saline and centrifuging at 9000 rpm for 10 minutes, and 400+ ml of distilled water was added to the resulting hemoglobin fraction and stirred at 5° C. for 90 minutes. After adding 2% hydrochloric acid to the obtained hemolysate to adjust the pH to 2.1, 100+ ol of a 1% aqueous sodium polyacrylate solution was added while stirring, and stirring was continued for about 30 minutes. After separating the aggregates formed into LL by this treatment by filtration, 2% sodium hydroxide solution iB was added to the filtrate containing globin to make I) 1 (6.4), and the protein was precipitated to isoelectrically. The precipitate was collected by centrifugation, washed with t15M-sodium phosphate buffer 1[V5It (1+sl), and the precipitate was centrifuged twice and frozen. No. 7. The dried globin obtained by drying was light yellowish brown in color, yielding 42 g,
The total nitrogen content by the Kerbourg method was 11.1% (71.3% as the parent protein), and the results of heme confirmation for the second globin using the benzunone reagent method and the pyrinone hemochrome method were negative. there were. Example 2 Fresh semi-coagulated blood (hemoglobin 9.7% 1000 ml)
In the same manner as in Example 1, 5 ot) sl of hemoglobin IB solution was obtained using 111jL. After 2% hydrochloric acid was added to this solution to adjust the pH to 2.3, 250 ml of a 1% aqueous sodium polyacrylate solution was added with stirring, and the reaction was continued for about 30 minutes. The aggregates generated during this treatment were separated by filtration, and 5 g of activated carbon was added to the filtrate and heated to 50°C.
2. After continuing to stir for 1 hour, it was filtered to obtain a pale yellow liquid containing globin. Then, through this filtration, 2%
- Add sodium hydroxide aqueous solution to adjust the pH to 6.7,
Proteins are precipitated isoelectrically. The precipitate was diluted by centrifugation I) and washed with TlH6,SQ,05M-sodium citrate buffered gson 1. The dried globin obtained by centrifuging again and freeze-drying the precipitate was pale yellow with no blood odor, and the yield was 69.8 g.
The total nitrogen by Körbourg method was 11.8% (73.8% as crude protein).

Claims (1)

[Claims] Hemoglobin is separated from blood collected from animals, the obtained hemoglobin is made into an aqueous solution, and the pH of the aqueous solution is adjusted to 3.
．． 1. A method for producing a globin protein, which comprises dissociating hemoglobin into heme and globin by dissociating hemoglobin to 0 or less, and then adding polyacrylic acid or a salt thereof to aggregate and remove heme.