JP2005261288A - Food and drink containing 7s protein derived from kidney bean - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a neutral or acid vegetable protein material stably usable in a wide salt concentration range, to provide food and drink containing neutral or acid protein including the protein, to provide a neutral or acid protein material having excellent emulsifying ability in a wide salt concentration range, and to provide neutral or acid emulsified food of various salt concentration. <P>SOLUTION: This neutral or acid vegetable protein material is made stably usable in a wide salt concentration range through using 7S protein derived from kidney bean. The neutral or acid food and drink containing the protein material is also provided. The neutral or acid protein material can demonstrate excellent emulsifying ability in a wide salt concentration range. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a neutral to acidic food or drink containing a protein derived from kidney beans, and a vegetable emulsifier composition containing the protein as an active ingredient.

  Plant protein is used in various processed foods as a protein source excellent in nutritional physiological function or as a natural functional agent having functions such as emulsifying power, foaming power and binding power. Proteins derived from soybeans and wheat are actually produced, and separated soybean proteins are the most frequently used. However, the solubility of these vegetable proteins is generally greatly affected by pH and salt concentration (ionic strength), and in particular in the acidic range, the solubility is poor and precipitation and aggregation occur. It was difficult to use as an agent or as a protein source for food and drink.

Various measures have been taken to solve these problems.
For example, the most commonly used means for preparing acidic protein beverages is to improve the acid solubility of the protein, but to add a stabilizer to stably disperse the protein. It is. Patent documents 1, 2, 3, etc. correspond to this, and vegetable protein is also object. However, these techniques cannot provide functions such as emulsifying ability.
Alternatively, it is common practice to perform enzymatic degradation of the protein in order to improve the acidic functionality of the protein. Patent Document 4 discloses a method for producing a mayonnaise-like food that improves acid emulsification by enzymatic decomposition of soybean protein, and is emulsified by adding oil, fat, salt, seasoning, and the like. . However, many problems remain, such as bad taste such as bitterness caused by decomposition, and insufficient emulsifying power.

JP 59-151837 JP-A-60-262584 JP-A-62-111632 JP 56-26171 A

  This invention makes it a subject to provide the protein raw material which has functions, such as favorable solubility and emulsification ability not only in neutral but in acidity. It is another object of the present invention to provide a neutral or acidic food or drink containing the protein material and an emulsifiable food or drink that has undergone an emulsification step.

As a result of intensive studies on the above problems, the present inventors have found that the kidney bean-derived 7S protein is neutral to acidic and exhibits good solubility in a wide range of salt concentrations, and has excellent emulsifying ability. The present invention was completed by discovering that it can be used effectively, and that it can be suitably used as a protein material and as an emulsifier composition in foods and drinks and the like without requiring pretreatment such as decomposition and addition of a stabilizer.
That is, the present invention
1. Food and drink containing 7S protein derived from kidney beans,
2. Acidic food and drink containing 7S protein derived from kidney beans,
3. The food or drink according to 1 or 2, which has undergone an emulsification step,
4). Emulsifier composition containing 7S protein derived from kidney beans as an active ingredient,
It is about.

  The present invention provides a protein material containing 7S protein derived from kidney beans as an active ingredient, and a food and drink containing the material, which are neutral to acidic and can be stably used in a system having a wide range of salt concentrations. Is. Moreover, the emulsifier composition which uses 7S protein derived from a kidney bean as an active ingredient, and the emulsifiable food / beverage products which passed through the process of the emulsification containing this emulsifier composition are provided.

Isolated soybean protein is most often produced as a vegetable protein and is used in various foods because of its nutritional value and functional properties. The main uses are sausages, hams, fishery products, etc., and functional properties such as water retention, oil retention and gelation are used. However, the separated soybean protein is insoluble in acidity, so it is difficult to use it in foods and drinks, and functions such as emulsifying power are also significantly reduced. For these reasons, use is limited to neutral foods such as sausages and hams, and acidic emulsified foods such as mayonnaise cannot be obtained.
The kidney bean-derived 7S protein in the present invention is not limited to being neutral but is acidic and is dissolved in a wide range of salt concentrations, exhibits functions such as emulsifying ability, and can be used stably. Therefore, even an acidic beverage, a food such as jelly, and an emulsifying food such as mayonnaise can be prepared, and the use is not limited to a neutral food or drink. Details will be described below in order.

  In the present invention, the kidney bean-derived 7S protein is a fraction containing a large amount of 7S globulin, which is a major component of the storage protein of kidney beans. The production method may be any conventionally known method, and fractionation by salting out, fractionation by pH adjustment, or a combination thereof may be employed. Preferably, the protein is fractionated after extracting the protein after degreasing.

  For example, ground kidney beans are defatted with a polar solvent such as hexane, extracted with a phosphate buffer (pH 7.6, 0.4 M NaCl) containing a reducing agent, and then a protein other than 7S globulin in 55% saturated ammonium sulfate. It is obtained by precipitating and removing 7S protein rich in 7S globulin in 80% saturated ammonium sulfate.

  The kidney bean-derived 7S protein in the present invention contains 70% by weight or more as 7S globulin, and preferably 7% globulin in the 7S protein is preferably 80% by weight or more, more preferably 90% by weight or more. A method for measuring the content of 7S globulin in the 7S protein will be described later.

  The food and drink in the present invention does not matter the content of 7S protein derived from kidney beans, but in the case of foods other than beverages, it generally contains 0.001 to 30% by weight of the protein, and in the case of beverages, it contains 0.001 to 10% by weight. Is preferred. In the case of a beverage, if it is too much, the viscosity increases and the drinking mouth becomes heavy. Conventionally, many neutral beverages containing soy protein like soy milk are found, but few are acidic. As mentioned above, this is acidic and slightly soluble in soy protein and requires the addition of a stabilizer, but it is still insufficient in stability and has a poor quality and a good quality. It is hard to get. However, the kidney bean-derived 7S protein in the present invention exhibits excellent solubility even in the acidic region, and therefore, it is possible to prepare a beverage with good quality without requiring a stabilizer or the like. However, the food and drink in the present invention does not prevent the use of the stabilizer.

  The food and drink in the present invention are not particularly limited in pH and salt concentration. However, if the protein concentration is adjusted within the above range, it can be used in a dissolved state, unlike a separated soybean protein, in a wide range of salt concentrations not only in neutral but also in acidity. Specifically, at pH 6.5 or higher or pH 5.0 or lower, protein can be contained in a dissolved state in a wide range of ionic strength 0.01 to 1.5. Even if the food or drink is in a pH range other than that, that is, lower than pH 6.5 and higher than pH 5.0, the protein can be contained in a dissolved state by adjusting the salt concentration to a range of 0.04 to 1.5. It is preferable that the protein is dissolved because it is excellent in throat, texture and aesthetics.

  The emulsifier composition in the present invention contains the above 7S protein containing 70% by weight or more as 7S globulin as an active ingredient, and is preferably 80% by weight or more, more preferably 90% as 7S globulin in the 7S protein. % By weight or more.

  The emulsifier composition in the present invention has acid resistance, and can be used well by adjusting the salt concentration even in an acidic region where other protein materials aggregate in addition to neutrality. For example, not only emulsifying foods such as neutral ice creams, hams and sausages, but also acidic emulsifying foods such as mayonnaise can be prepared. Specifically, when the pH is 6.5 or more or pH 5.0 or less, good emulsifying power is exhibited in a wide range of ionic strength I = 0.01 to 1.5. Moreover, even if the pH is higher than 5.0 and lower than pH 6.5, if the ionic strength I is 0.05 to 1.5, the same good emulsifying properties are exhibited.

  The ionic strength is an indicator of the salt concentration in the system. A system having an ionic strength of 0.04 corresponds to a sodium chloride aqueous solution of about 0.2%, and a system having an ionic strength of 0.5 corresponds to a sodium chloride aqueous solution of about 2.9%. The salinity of various emulsifying foods is about 0.2% for ice cream, 1.8% for mayonnaise, 2.3% for egg yolk, and 2 to 3.8% for cheeses, although it varies depending on the type. Food Composition Table, Women's Nutrition University Press). However, since the value of these food products is the salt concentration relative to the total weight of the water phase and the oil phase, the salt concentration in the aqueous phase is higher than this. The emulsifier composition in the present invention can be used for a wide range of emulsifiable foods and drinks from foods having almost no salty taste to strong foods.

  The emulsifiable food / beverage products in this invention contain fats and oils and aqueous solution other than the said emulsifier composition. The aqueous solution is a solution containing water as a main component and is not particularly limited as long as it is edible. As the fats and oils, any fats and oils can be used, and are generally fats and oils made from animals, plants and microorganisms which are edible oils. For example, lard, beef tallow, chicken oil, whale oil, tuna oil, sardine oil, mackerel oil, saury oil, bonito oil, herring oil, liver oil, soybean oil, cottonseed oil, safflower oil, rice oil, corn oil, rapeseed oil, Oils and fats blended with palm oil, perilla oil, sesame oil, cacao butter, peanut oil, coconut oil and the like and synthetic triglycerides such as medium chain fatty acid triglycerides, and these can be used alone or in appropriate combination.

  In the present invention, the content of fats and oils is appropriately 0.0005 to 70% by weight, preferably 0.001 to 60% by weight, more preferably 0.003 to 50% by weight in the emulsified food or drink. If it is this range, the fat-containing water-soluble composition which has practical value can be obtained, and fats and oils can be easily emulsified or solubilized.

  The content of the emulsifier composition in the emulsifiable food and drink according to the present invention depends on the fat and oil content, pH, and ionic strength, but as the kidney bean 7S globulin, 0.005 to 5% by weight, preferably 0.03 to 3% by weight, Within this range, a good emulsified state can be obtained.

  The emulsifiable food and drink in the present invention can be obtained by emulsifying fats and oils and an aqueous solution. The emulsification method may be any conventionally known method, but a colloid mill, a high-pressure homogenizer, a microfluidizer, and a nanomizer. Or a homogenization processor such as an ultrasonic emulsifier can be used. Moreover, the sterilization process by a conventional method can be performed as needed.

  The emulsifiable food and drink in the present invention includes all foods that are emulsified with the emulsifier composition in the present invention. For example, seasonings (mayonnaise, dressing, sauce, etc.), flour products (bread, cake, Noodles, batter mix, etc.), livestock products (ham, sausage, hamburg, meatballs, etc.), frozen dessert (ice cream, sherbet, etc.), dairy products (butter, cheese, yogurt, etc.), processed fats and oils (margarine, fat) Spread, shortening, etc.), confectionery (Western confectionery, Japanese confectionery, etc.), dessert food (jelly, pudding, etc.), beverages, tofu, frozen food, side dishes, etc. There are a wide variety of tablets, powders and granules.

The analysis method used in the present invention is described below. Added a method for measuring protein concentration.
* Protein concentration: It was measured using a protein assay rapid kit manufactured by Wako Pure Chemical Industries, Ltd.
* SDS-PAGE: Based on the method of Laemmli (Nature. 227, 680 (1970)), analysis was performed at a gel concentration of 11%. The amount of sample applied was 10 μg.
* Globulin content of each protein: The migration pattern obtained by SDS-PAGE was measured with a densitometer, and the area ratio of the corresponding fraction relative to the whole was defined as purity.

<Production example>
1 L of hexane was added to 100 g of ground kidney beans, stirred for 1 hour at room temperature, and allowed to stand for 30 minutes, and the supernatant was removed. To this was added 1 L of acetone, and the mixture was stirred at room temperature for 1 hour and allowed to stand for 30 minutes to remove the supernatant. 1 L of diethyl ether was added thereto, and the mixture was similarly stirred at room temperature for 1 hour and allowed to stand for 30 minutes. The supernatant was removed, and the precipitate was dried to obtain a defatted kidney bean powder. To 50 g of this powder, 1 L of an extraction buffer containing 0.4 M NaCl (pH 7.6, 35 mM sodium phosphate, 1 mM EDTA, 0.02% NaN3, 10 mM 2 mercaptoethanol) was added, stirred at room temperature for 2 hours, and centrifuged (16000 rpm, 30 minutes) to obtain a supernatant fraction containing the protein extract. To this was added ammonium sulfate to 55%, and the mixture was stirred on ice for 30 minutes and centrifuged (16000 rpm, 30 minutes, 4 ° C.) to recover the supernatant fraction. Ammonium sulfate was added to the supernatant to 80%, and the mixture was stirred again on ice for 30 minutes and centrifuged (16000 rpm, 30 minutes, 4 ° C.) to remove the supernatant, and about 5 g of 7S protein was recovered. This was eluted with 100 ml of extraction buffer. The 7S globulin content of the 7S protein was 90%.

<Comparative Production Example 1>
To 100 g of defatted soybean, 15 volumes of 30 mM pH 8.0 Tris-HCl buffer (1 mM EDTA) was added, and extraction was performed at room temperature for 30 minutes. Centrifugation was performed (9,200 g, 20 minutes), and okara was separated and removed to obtain about 1200 g of a soybean protein extract solution as a supernatant. The protein concentration in the soy protein extract obtained was 3.1% by weight.

<Comparative Production Example 2>
15 times the amount of water was added to 100 g of defatted soybeans, adjusted to pH 7.5 with sodium hydroxide, and extracted at room temperature for 30 minutes. Centrifugation (9,200 g, 20 minutes) was performed to separate and remove okara. Sodium bisulfite was added to the obtained protein extract to 10 mM, and the pH was adjusted to 6.4 with hydrochloric acid. This was left at 4 ° C. overnight and centrifuged (6760 g, 20 minutes) to obtain a precipitate fraction consisting of 11S protein. Sodium chloride was added to the supernatant fraction to 0.25 M, the pH was adjusted to 5.0 with hydrochloric acid, and the mixture was centrifuged again (6760 g, 20 minutes) to obtain a supernatant fraction. This was diluted 2-fold with water, adjusted to pH 4.5 with hydrochloric acid, and centrifuged (6760 g, 20 minutes) to recover the 7S protein as a precipitate fraction. The yield of 11S protein was about 10 g and the content of 11S globulin was 94%. The yield of 7S protein was about 6 g and the content of 7S globulin was 95%.

<Reference Example 1>
The pH dependence of solubility at an ionic strength of 0.08 of 7S protein derived from kidney beans obtained in Production Example was examined. For comparison, 7S and 11S proteins derived from soybean of Comparative Production Example 2 were used. Each protein was dialyzed against 10 mM NaPi buffer (ionic strength 0.53) of pH 7.6 containing 0.5 M NaCl, and then the protein concentration was adjusted to 8 mg / ml.
Also, 10 mM Macllvaine buffer at pH 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 10 mM NaPi buffer at pH 5.5, 6.5, 8.0, 10 mM NH ₄ Cl at pH 8.0, 9.5, 11.0 Each buffer (ionic strength 0.03) of -NH ₄ OH buffer was prepared.
Each of these buffers and the protein solution were mixed in an eppen at a ratio of 9: 1. That is, the pH and ionic strength of the final protein solution were adjusted by diluting the protein solution with each buffer. This was allowed to stand at 4 ° C. for 18 hours, and then centrifuged at 15,000 rpm at 4 ° C. for 15 minutes to recover the supernatant.
The final pH and protein amount of each supernatant were measured, and the solubility at each pH was determined. However, the one with the highest protein content was defined as 100% solubility. The results are shown in FIGS. As a result, even in the pH range where soybean protein is insolubilized, 7S protein derived from kidney beans is dissolved, indicating that it is an emulsifier that can be used in a wide range of acidic to neutral.

<Reference Example 2>
The dependence of the 7S protein derived from kidney beans obtained in the production example on the ionic strength at each pH was investigated. After dialyzing the kidney bean-derived 7S obtained in the production example, 10 mM NaPi buffer (ionic strength 0.03) at pH 7.6, 7.0, 6.5, 6.0, 5.5, 5.0 and 10 mM NaPi buffer (ionic strength 0.08) containing 0.05 M NaCl The protein concentration was adjusted to 8 mg / ml.
Separately, NaPi buffers having ionic strengths of μ = 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.008, and 0.005 with different concentrations of NaCl and NaPi were prepared. The ionic strength of the final protein solution was adjusted by diluting the above protein solution with these buffers.
Specifically, a protein solution with an ionic strength of 0.08 at pH 7.6, 7.0, 6.5 is used for a solubility buffer with an ionic strength of 0.08 to 0.03, and a protein solution with an ionic strength of 0.03 is used for a solubility with an ionic strength of 0.03 to 0.005. The buffer was mixed with Eppen at a ratio of 1: 9. In addition, protein solutions having an ionic strength of 0.08 at pH 6.0, 5.5, and 5.0 were mixed with a solubility buffer having an ionic strength of 0.08 to 0.005 in an eppen at a ratio of 1: 9.
The final ionic strength of the mixed solution thus obtained was ionic strength 0.08, 0.071, 0.062, 0.053, 0.044, 0.035, 0.03, 0.021, 0.013, 0.01, 0.008 for pH 7.6, 7.0, and 6.5, respectively. . Moreover, pH 6.0, 5.5, and 5.0 were ionic strength 0.08, 0.071, 0.062, 0.053, 0.044, 0.035, 0.026, 0.017, 0.015, and 0.013, respectively. This was allowed to stand at 4 ° C. for 18 hours, and then centrifuged at 15,000 rpm at 4 ° C. for 15 minutes to recover the supernatant. The amount of protein in each supernatant was measured, and the solubility at each ionic strength was determined. However, the one with the highest amount of protein was defined as 100% solubility. The results are shown in FIGS. From this result, it was shown that 7S protein derived from kidney beans has good solubility even in a pH range where the protein derived from soybean is insoluble.

<Experimental example>
The emulsion stability of 7S protein derived from kidney beans obtained in Production Example at pH 7.6 was examined. For comparison, soybean protein, 7S protein, and 11S protein obtained in Comparative Production Example 1 or 2 were used.
Each protein was dissolved in pH 7.6 phosphate buffer (ionic strength 0.08) so that the protein concentrations would be 0.125 mg / ml and 0.25 mg / ml. 0.25 ml of soybean oil and 1.5 ml of each protein solution are first used in a Hiscotron ultra-high speed homogenizer (Nisson Medical Science Instruments) (27,000 rpm, 30 seconds), followed by an ultrasonic homogenizer (Nippon Seiki Seisakusho) Homogenization was performed at 100 μA for 1 minute to prepare an emulsion. The state after 20 hours of the obtained emulsion was observed to evaluate the emulsion stability. The kidney bean 7S protein was well emulsified at any protein concentration and was stable after 20 hours. On the other hand, although all the proteins derived from soybean were emulsified, they were unstable and completely separated after 20 hours.
Subsequently, the emulsion stability of 7S protein derived from kidney beans obtained in Production Example at pH 6.0 was examined. The protein was dissolved in a pH 6.0 phosphate buffer (ionic strength 0.06) to a protein concentration of 0.25 mg / ml and emulsified in the same manner to obtain a good emulsion. On the other hand, soybean-derived 11S protein and 7S protein were both insoluble and could not be emulsified at all.

  A vegetable mayonnaise-like food was prepared using the 7S protein obtained in Production Example. 6 g of 7S protein, 1 g of acetic acid, 5 g of sodium chloride and 1 g of glutamisan soda were dissolved in 87 g of water, and 150 g of salad oil was dispersed and emulsified with a homogenizer (EXCEL AUTO HOMOGENIZER manufactured by Nippon Seiki Co., Ltd.). The oil was added in several batches and emulsified for 1 minute at 10,000 rpm each time. Mayonnaise (pH 4.0) with good emulsification and shape retention was obtained, and the flavor was also good. It has been shown that a good emulsion can be prepared even in an acidic and salt-containing system.

Solubility of kidney beans-derived 7S protein (pH 7.6, ionic strength 0.08) Solubility of 11S protein derived from soybean (pH 7.6, ionic strength 0.08) Solubility of 7S protein derived from soybean (pH 7.6, ionic strength 0.08) Solubility of 7S protein derived from kidney beans (pH 7.6) Solubility of 7S protein derived from kidney beans (pH 7.0) Solubility of 7S protein derived from kidney beans (pH 6.5) Solubility of 7S protein derived from kidney beans (pH 6.0) Solubility of 7S protein derived from kidney beans (pH 5.5) Solubility of 7S protein derived from kidney beans (pH 5.0)

Claims (4)

Food and drink containing 7S protein derived from kidney beans. Acidic food and drink containing 7S protein from kidney beans. The food or drink according to claim 1 or 2, which has been subjected to an emulsification step. An emulsifier composition comprising 7S protein derived from kidney beans as an active ingredient.

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