JP2006061016A - Method for developing color of meat product - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for developing color of a meat product, by which the meat product can maintain its vivid red color for a long period of time without causing toxicity in itself and producing carcinogenic substance. <P>SOLUTION: The method for developing color of a meat product comprises making a ferrochelatase-containing material come in contact with meat preferably at 40-60°C. In such a process, the ferrochelatase-containing material preferably comprises mitochondrial fractions such as those obtained from at least one kind selected from mammals, birds and fish, or those obtained from at least one kind selected from filamentous fungi, yeast and bacteria. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a meat product coloring method, and more particularly to a meat product coloring method that does not use nitrites that are suspected of producing carcinogenic substances.

  The red color of livestock meat such as beef, pork, chicken, lamb, and horse meat or fish such as salmon and salmon is browned when exposed to the air, causing a significant reduction in commercial value. It is known that the color of such red meat changes due to myoglobin and hemoglobin, and changes in color tone such as browning and fading when bright red myoglobin and hemoglobin are oxidized. In particular, in processed meat products such as ham and sausage, nitrite and nitrate are added to give a beautiful color to produce a beautiful color from heating raw meat, L-ascorbic acid, ersorbic acid, A method of preventing discoloration by adding nicotinamide or the like is generally performed. In addition, a method of adding kojic acid and sodium L-ascorbate [see Patent Document 1], a method of causing leucoanthocyanidin, proanthocyanidin, leucoanthocyanidin or proanthocyanidin and ascorbic acid (salt) to act [see Patent Document 2], A method using a pigment extracted from western red rape (see Patent Document 3), a composition component of deep sea water, and further adding a rosemary extract thereto (see Patent Document 4), a method of increasing the pH of the surface layer [Patent Document 3] Many methods have been proposed, such as a method in which yeast is added and iron porphine in animal meat is substituted with zinc to develop color (see Patent Document 6).

Japanese Patent Publication No. 5-25463 JP-A-10-004923 JP 05-015343 A JP 2003-093016 A JP 07-213253 A Japanese Patent Application No. 2003-323087 (filed on September 16, 2003) Specification

Although the methods using the above-mentioned dyes or the methods for preventing oxidation have their respective effects, the color tone is not sufficiently stable, and at present the use of making a stable binding state with myoglobin using nitrite is used. The method is still the main force. However, nitrite is particularly toxic among food additives, and it has been pointed out that it may combine with amino acid degradation products to form carcinogenic nitrosamines. Therefore, there is a strong demand for a method for retaining red meat that is safe for the human body.
From such a viewpoint, an object of the present invention is to provide a method that is not toxic in itself, does not produce a carcinogenic substance, and can keep a meat product in a bright red color for a long time.

  In order to solve this problem, the meat product coloring method according to claim 1 is to bring a ferrochelatase-containing material into contact with meat.

  In the meat product coloring method according to claim 2, the ferrochelatase-containing material according to claim 1 is a mitochondrial fraction.

  In the method for coloring meat products according to claim 3, the mitochondrial fraction in claim 2 is a fraction from one or more animal internal organs selected from mammals, birds and fish.

  In the meat product coloring method according to claim 4, the mitochondrial fraction in claim 2 is a fraction from one or more selected from filamentous fungi, yeasts, and bacteria.

  The meat product coloring method according to claim 5 is that the ferrochelatase-containing material according to claim 1 is brought into contact with meat at 40 to 60 ° C.

  As an effect of the present invention, it is possible to give a good color tone to meat products by a method safe for the human body without using nitrite suspected of causing carcinogenicity.

  The meats targeted by the present invention are livestock meat such as beef, pork, horse meat, lamb, goat meat, poultry meat, and rabbit meat, and also include whale meat and fish such as salmon and salmon.

  The fact that the meat is bright red is due to the myoglobin in the meat. Myoglobin is composed of globin protein and iron protoporphine, and changes its color by browning or fading due to oxygenation in the air. In the patent document 6, the present inventors have already replaced this iron with zinc to obtain a myoglobin zinc protoporphine IX complex (hereinafter referred to as “myoglobin / heme zinc”), whereby the bright red color of meat is Although it has been reported that the stability to oxidation and heating is significantly improved and a good color tone can be maintained for a long period of time, the present invention promotes the formation of myoglobin and heme zinc in meat by contacting the meat with ferrochelatase. This is a method of coloring meat. Also, hemoglobin in blood is exactly the same, and hemoglobin can similarly substitute iron in hemoglobin with zinc to significantly improve the stability against oxidation and heating, and give good color tone. Hereinafter, the present invention will be described by using myoglobin as a representative.

  The present invention is characterized by accelerating the action of replacing the iron in the heme group of myoglobin with zinc by the action of a ferrochelatase-containing substance. Here, myoglobin / heme zinc has the following chemical structure.

  Ferrochelatase is one of the enzymes involved in heme synthesis, and is particularly localized in animal tissues, plant tissues, yeast, bacteria, and the like. For example, in animal tissues, ferrochelatase is present in almost all parts of mammals, birds, fish, etc., but as described in the Examples, it is contained more in the internal organs than in the muscle parts. Considering the color development, it is preferable to use visceral parts such as kidney, liver, spleen and heart of meat animals. Other bacteria such as baker's yeast, beer yeast, sake yeast, wine yeast, shochu yeast, filamentous fungi for cheese production, filamentous fungus for tempeh production, natto fungus for natto production, mushrooms (shimeji , Maiko, shiitake mushrooms, eringi, enoki, etc.), bean sprouts, peas, etc. contain a large amount of ferrochelatase, and these can be used.

  The ferrochelatase should be extracted and purified from these, but the object can be achieved if it contains ferrochelatase. In practice, it is advantageous to use a mitochondrial fraction rich in ferrochelatase. . The mitochondrial fraction can be obtained by finely cutting the material containing a large amount of ferrochelatase and centrifuging it in an aqueous solution adjusted to pH.

  The zinc source for forming myoglobin and heme zinc is zinc that is originally contained in meat. The amount of zinc contained in the meat is approximately 2 to 6 mg / 100 g, which is approximately three times that of iron, although it varies depending on the type of animal and the portion of the meat. Although zinc may be added from outside, it is also effective to add a natural product rich in zinc, such as oyster meat extract, from the current situation that zinc is not currently recognized as a food additive.

  When a substitution reaction with myoglobin and heme zinc is performed in meat, the presence of phosphate, pyrophosphate, or polyphosphate is effective in promoting the reaction. The phosphates found in food additives are phosphoric acid, tripotassium phosphate, tricalcium phosphate, diammonium hydrogen phosphate, ammonium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, Calcium monohydrogen phosphate, calcium dihydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, trisodium phosphate, and pyrophosphates recognized in food additives are tetrapotassium pyrophosphate, pyrolin Calcium dihydrogen, disodium dihydrogen pyrophosphate, tetrasodium pyrophosphate, and polyphosphates recognized in food additives are potassium polyphosphate and sodium polyphosphate. The above is selected. The amount added is usually about 0.01 to 0.5% by weight based on meat.

  In the meat treatment with the ferrochelatase-containing material in the present invention, the ferrochelatase-containing material, preferably the mitochondrial fraction, is brought into contact with meat by adjusting the pH by adding a buffer solution as necessary. The contact with meat is performed by a method of injecting a liquid containing ferrochelatase in the case of block meat and tumbling, or mixing with the liquid in minced meat. Here, the amount of the ferrochelatase-containing material varies depending on the nature, form, content of the ferrochelatase, the type of meat, the shape of the meat, other processing conditions, etc., but generally 0.01 to It is 10% by weight, preferably 0.1 to 3% by weight.

  The purpose of the meat treatment can be achieved by bringing it into contact with the ferrochelatase-containing material and taking a sufficiently long time at room temperature. For example, it was confirmed that a substantially effective amount of myoglobin / heme zinc was produced when held at 37 ° C. for 5 days. However, holding at 37 ° C. for a long time has another problem such as meat rotting, and it is preferable to set the temperature to 40 to 60 ° C. so that it can be processed in a short time. Conversely, it can be carried out at a low temperature of 10 ° C. or less while avoiding meat rot, and in this case, it can be carried out simultaneously with the aging of the meat, and the object of the present invention can be achieved in 7 to 14 days.

  The processed meat is usually heated to 70 to 80 ° C. to be a meat product such as ham or sausage. Non-heated food such as prosciutto is stored at a low temperature of 10 ° C. or lower to become a product.

[Correlation between ferrochelatase activity and zinc substitution]
Ferrochelatase activity was compared for animal meat, viscera and yeast.
(1) Measurement of protein mass:
Each part of pork heart, kidney, liver, spleen, and pork loin, pork thigh, and beef thigh with the removal of fat and connective tissue are cut into small pieces with a food processor, each to 10g. 40 mL of 10 mM Tris-HCl buffer solution (pH 7.6) was added, and homogenized at 15,000 rpm for 2 minutes with ice-cooling using a homogenizer [manufactured by Nippon Seiki Seisakusho, “ACE Homogenizer AM-11” (model number)]. did. After removing large suspended solids through the mesh, the protein concentration of 1 mg of the mixture was measured. Measurement was performed using a Bradford method measurement kit manufactured by Bio Rad.
In addition, 10 g of dry baker's yeast was pulverized to about 2 μm with a multi-sample cell disrupter [manufactured by Yasui Kikai Co., Ltd., “Multi-Beads Shocker-MB-200W” (trade name)], and added to 90 mL of the buffer. And the same processing.

(2) Measurement of ferrochelatase activity 10 mg-Tris-hydrochloric acid (pH 7.6) homogenized product of each part of the above porcine heart, kidney, liver and spleen, and pork loin, pork thigh and beef thigh meat 2 mg / protein The sample solution was made up to mL. To 250 μL of this sample solution, 1 mM-deuteroporphyrin IX (manufactured by Funakoshi Co., Ltd.) 7.5 μL, 1 M-Tris hydrochloric acid 50 μL, 10% -Tween 80 (nonionic surfactant) 1.5 μL, 1 mM-CoSO _4. 7 H ₂ O [manufactured by Wako Pure Chemical Industries, Ltd.] 1.5 μL and pure water 26 μL were added and reacted at a predetermined temperature for 30 minutes. Before and after the reaction, a 150-fold (volume) amount of 80% acetone was added and mixed to stop the reaction, and then the spectrofluorophotometer ["RF-5000" manufactured by Shimadzu Corporation (model number) )]. Measurements were excited at 400 nm and fluorescence was recorded at 620 nm, with each slit having excitation at 5 nm and fluorescence at 10 nm. In addition, a calibration curve was prepared with deuteroporphyrin IX separately dissolved in 80% acetone. The amount of deuteroporphyrin IX decreased from the absorbance of the sample solution and the calibration curve, and the amount of deuteroporphyrin IX (pmol) at which 1 mg of protein introduces Co in one minute was calculated.

The results are shown in Table 1.

(3) Zinc replacement activity 10 mM-Tris-HCl (pH 7.6) homogenized product (with a protein concentration of 10 mg / mL) of the above porcine heart, kidney, liver, spleen, and pork loin, pork thigh and beef thigh Preparation) 30 parts by weight, zinc gluconate 0.1 part by weight, equine myoglobin [Sigma Aldridge Japan, Reagent] 0.1 part by weight, dithiothreitol 0.03 part by weight, 0.2 M phosphate buffer (pH 5) .5) 69.77 parts by weight were mixed with a homogenizer, and lightly stirred at 37 ° C. for 24 hours and 45 ° C. for 2 hours to replace zinc. The zinc substitution rate is the ratio of the amount of myoglobin / heme zinc to the total amount of myoglobin / heme zinc and iron, and the amount of myoglobin / heme zinc and the total amount of iron is obtained from the absorbance at 383 nm, and the amount of myoglobin / heme zinc is obtained from the absorbance at 417 nm. It calculated | required by calculation from each value. The results are shown in Table 2.

この結果から、表１のフェロケラターゼ活性度と表２の亜鉛置換活性は、非常によく相関していることがわかる。

From this result, it can be seen that the ferrochelatase activity in Table 1 and the zinc substitution activity in Table 2 correlate very well.

[Preparation of mitochondria from pig heart]
(1) Preparation of mitochondria from pig heart As long as fat and connective tissue were removed from the fresh pig heart, it was cut into small pieces with scissors. This was put in 0.2 M phosphate buffer solution (pH 6.0) of 10 times weight, and using a homogenizer [manufactured by Nippon Seiki Seisakusho, “ACE Homogenizer AM-11” (model number)], 4 ° C., 15 Homogenized at 1,000 rpm for 30 seconds. The homogenized solution was placed in a centrifuge tube and centrifuged at 960 × g and 0 ° C. for 7 minutes. The centrifuged supernatant was put in a centrifuge tube and centrifuged at 17400 × g and 0 ° C. for 12 minutes to obtain mitochondria as a precipitate.

(2) Preparation of mitochondria from baker's yeast I. Resaffle (S.I.Lesaffle), "Saf-Instant Red" (trade name)], a multi-sample cell crusher [Yasui Kikai Co., Ltd., "Multi-Bead Shocker MB-200W" (Trade name)] is crushed to about 2 μm, placed in a 10-fold weight 0.2 M phosphate buffer solution (pH 6.0), and homogenizer [same as above] at 4 ° C., 15 Homogenized at 1,000 rpm for 30 seconds. The homogenized solution was placed in a centrifuge tube and centrifuged at 900 × g and 0 ° C. for 30 minutes. The centrifuged supernatant was put in a centrifuge tube and centrifuged at 40000 × g and 0 ° C. for 30 minutes to obtain mitochondria as a precipitate.

[Separation of ferrochelatase from mitochondria]
(1) Separation of ferrochelatase from baker's yeast mitochondria (a) The baker's yeast mitochondria fractionated in Example 3 contains 0.1 mM-EDTA (ethylenediaminetetraacetic acid) and 0.1 mM-PMSF (phenylmethanesulfonyl fluoride). The protein was added to and suspended in a 0.1 M potassium phosphate buffer solution (pH 7.6) so that the protein concentration was 50 mg / mL.
(B) To this suspension, a nonionic surfactant [manufactured by Wako Pure Chemical Industries, Ltd., “polyoxyethylene monooleate”] was added to a protein mass ratio of 0.5 (weight ratio). It was dissolved in 1M potassium phosphate buffer solution (pH 7.6) and mixed.
(C) Using ultrasonic treatment (“Sonicator” (trade name) manufactured by Otake Seisakusho Co., Ltd.), the treatment was performed three times at 100 WAT at 4 ° C. for 1 minute, and further at 4 ° C. with a magnetic stirrer. Stir for hours.
(D) Next, the mixture was centrifuged at 150,000 × g for 90 minutes, and glycerol was added to the supernatant to a final concentration of 20% by weight.
(E) This solution was equilibrated with a 25 mM Tris-HCl buffer solution (pH 8.0) containing 20% glycerol and 1% Tween 80 (nonionic surfactant) [Amersham Biosciences, “Blue -Sehalose CL6B column] (trade name), column diameter 2.6 × 40 cm ”.
(F) The resin was eluted with 0.1M potassium phosphate buffer solution (pH 7.6) containing 1M-KCl and 1% -cholate, and the eluate was dialyzed and lyophilized to obtain ferrochelatase. Obtained.
About the ferrochelatase isolate | separated here, it carried out similarly to Example 1, and measured ferrochelatase activity and zinc substitution activity. The results are shown in Table 3.

[Injection into meat]
(1) Preparation of sausage The mitochondrial fractions of Examples 2 and 3 and other treatment agents were added to pork urine minced meat, homogenized, and filled into a case with a folding diameter of 3 cm to obtain a model sausage. The mixture was reacted at 50 ° C. for 2 hours in a warm bath and sterilized by heating at 75 ° C. for 30 minutes. After cooling overnight, the zinc replacement rate and meat color were observed. The hue of the meat was measured using a color difference meter (“Color Tester” (trade name) manufactured by Suga Test Instruments Co., Ltd.). Table 4 shows the blending ratio and measurement results.

この結果からみられるように、肉にミトコンドリア画分を添加することにより発色が促進され、赤味のある色調に仕上げることができた。

As can be seen from this result, the color development was promoted by adding the mitochondrial fraction to the meat, and it was possible to finish it in a reddish color tone.

  INDUSTRIAL APPLICABILITY The present invention gives a good color tone without using nitrite suspected of causing carcinogenicity to meat, and since this color tone is stable for a long period of time, a processed meat product considering human health can be manufactured. .

Claims (5)

  A method for coloring a meat product, comprising bringing a ferrochelatase-containing material into contact with meat.   The meat product coloring method according to claim 1, wherein the ferrochelatase-containing material is a mitochondrial fraction.   The meat product coloring method according to claim 2, wherein the mitochondrial fraction is a fraction from one or more animal internal organs selected from mammals, birds and fish.   The meat product coloring method according to claim 2, wherein the mitochondrial fraction is a fraction from one or more selected from filamentous fungi, yeasts, and bacteria.   The method for coloring a meat product according to claim 1, wherein the ferrochelatase-containing material is brought into contact with meat at 40 to 60 ° C.