JP5541161B2 - Method for producing phosphorylated water-soluble polysaccharide - Google Patents

Description

  The present invention relates to a phosphorylated water-soluble polysaccharide having dispersion stability of protein particles, and a novel dispersion stabilizer for protein particles containing the polysaccharide as an active ingredient.

  Acidic milk beverages are known in which milk proteins are fermented with sugars or dispersed in an aqueous solution of organic acids. For this purpose, high methoxy pectin and carboxymethyl cellulose are used as dispersion stabilizers for milk protein particles. Has been. As a dispersion stabilizer, water-soluble soybean polysaccharide obtained by high-temperature pressure extraction under acidic conditions using okara from which oil and fat and protein are separated and removed from soybean as disclosed in Patent Document 1 is used. In addition, milk protein particles can be dispersed and stabilized in an extremely low viscosity state under an acidity of less than pH 4 that cannot be achieved with pectin or the like. This makes it possible to produce a refreshing and light-taste acidic milk beverage that is different from a heavy-taste beverage with a pasty feeling to which pectin is added (Patent Document 2).

  By the way, acidic milk drinks include those with high protein content and those made with fermented milk. If these beverages are prepared using water-soluble soy polysaccharides under a pH of less than 4, it will become a very sour beverage, or lactic acid bacteria cannot grow due to low pH, and a live type lactic acid bacteria beverage should be produced There is a problem that cannot be done. On the other hand, the above-mentioned water-soluble soybean polysaccharides cannot exhibit sufficient dispersion stabilizing ability of protein particles in the pH range exceeding pH 4.2, or if the amount added is increased, the refreshing mouth is not clean. It will interfere. Therefore, a water-soluble polysaccharide that can disperse and stabilize milk protein particles in a pH environment higher than pH 4.2 and low viscosity is desired.

  Milk proteins are positively charged under acidity below their isoelectric point, but research is progressing on models that disperse milk protein particles in acidic milk beverages with polysaccharides, especially pectin and water-soluble soybean polysaccharides. (Non-Patent Document 1). Polysaccharides that contribute to the dispersion stability of milk protein particles are ionically or hydrophobically bound to the protein surface by their own negative charges, thereby suppressing aggregation and precipitation of protein particles due to electrical repulsion and steric hindrance. It is believed that. To date, many attempts have been made to increase the negative charge of polysaccharides in order to improve the dispersion stability of the polysaccharides. For example, carboxymethyl cellulose (CMC) in which carboxymethyl groups are introduced into cellulose to increase the negative charge. However, CMC does not necessarily have a high dispersion stabilizing power of milk protein particles, and the viscosity of the beverage increases, making it difficult to obtain a refreshing beverage.

Japanese Patent No. 2599477 Japanese Patent No. 3280768

J. Agric. Food Chem., 54 (17), 6241 -6246, 2006.

  Disperse and stabilize protein particles even in the pH range above pH 4.2, which is near the isoelectric point of protein, where the stability of conventional water-soluble soybean polysaccharides is reduced. The purpose is to do.

  As a result of intensive research on the above problems, the present inventors dispersed milk protein particles in the pH range near the isoelectric point exceeding pH 4.2, which could not be stabilized by conventional water-soluble soybean polysaccharides. The present invention has been completed by finding that it is stabilized.

That is, the present invention
(1) A method for producing a phosphorylated water-soluble polysaccharide, which comprises treating a water-soluble polysaccharide having an acidic sugar as a constituent sugar with a divalent metal ion and a phosphate ion.
(2) The phosphorylated water-soluble polysaccharide according to (1), wherein the water-soluble polysaccharide having an acidic sugar as a constituent sugar is treated with a divalent metal ion and subsequently treated with a phosphate ion. A method for producing sugars.
(3) The production method according to (1), wherein the water-soluble polysaccharide is a soybean polysaccharide.
(4) The production method according to (1), wherein the divalent metal ion is derived from calcium or magnesium.
(5) The production method according to (1), wherein the phosphate ions are derived from one or more selected from phytic acid, metaphosphoric acid, and polyphosphoric acid.
(6) A phosphorylated water-soluble polysaccharide produced by the method according to (1) to (5).
(7) A dispersion stabilizer for protein particles comprising the phosphorylated water-soluble polysaccharide according to (6) as an active ingredient.
(8) An acidic protein food containing the phosphorylated water-soluble polysaccharide according to (6).
(9) A method for enhancing the protein particle dispersibility of a water-soluble soybean polysaccharide, characterized by treating the water-soluble soybean polysaccharide with a divalent metal ion and a phosphate ion.
(10) A method for enhancing the ability to disperse protein particles of a water-soluble soybean polysaccharide, characterized in that the water-soluble soybean polysaccharide is treated with a divalent metal ion and subsequently treated with a phosphate ion.
It is.

  ADVANTAGE OF THE INVENTION According to this invention, the protein is disperse-stabilized by pH exceeding pH4.2 which is the isoelectric point vicinity of milk protein with which the stabilization power was reduced conventionally, and the acidic milk drink of a drinking mouth with low viscosity is provided. be able to. In addition, it is possible to produce an acidic milk beverage in a pH range where lactic acid bacteria can grow, and an acidic milk beverage containing fermented milk of a live mouth type with low viscosity and a clean drinking mouth can be obtained.

(Water-soluble polysaccharide)
Hereinafter, the present invention will be specifically described. The water-soluble polysaccharide having an acidic sugar as a constituent sugar in the present invention is a water-soluble polysaccharide having an acidic sugar having a carboxyl group or a sulfate group as a constituent sugar. Examples of water-soluble polysaccharides whose acidic sugars have carboxyl groups include pectin obtained from root vegetables such as beets and potatoes, and fruits such as apples and citrus, and water-soluble soybean polysaccharides obtained from soybeans, alginic acid and the like. It is done. Moreover, various carrageenan etc. are mentioned as the water-soluble polysaccharide in which the acidic saccharide has a sulfate group. In the present invention, a water-soluble polysaccharide in which the acidic sugar has a carboxyl group is preferable, and a water-soluble soybean polysaccharide is particularly preferable because the dispersibility of protein particles is remarkably enhanced.

(Water-soluble soybean polysaccharide)
Although the present application can use water-soluble soybean polysaccharides obtained by various methods, for example, water-soluble soybean polysaccharides described in Patent Document 1 can be used. An example of the production is tofu, soy milk, and isolated soy protein produced as by-products in the production of okara and defatted soybean meal (meal) as raw materials, and the pH before extraction becomes 3-7 The extraction is carried out under conditions, preferably in the temperature range above 100 ° C. After extraction, solid-liquid separation is performed to obtain a supernatant. The obtained supernatant is preferably obtained by demethoxytreating methyl ester of galacturonic acid by a known method such as alkali treatment to enhance the dispersibility of protein particles.

(Purification)
The obtained water-soluble soybean polysaccharide can be used as it is, but it is desirable to carry out purification such as protein removal and desalting in order to exert more functions.
Protein removal methods include pH adjustment, protein aggregation, removal by pressure filtration, centrifugation, filtration, membrane separation, and other separation means, decomposition using any protease, activated carbon and resin. And a purification method for adsorbing and removing impurities. As a method for desalting, any method capable of separating and removing salts may be used. Examples thereof include precipitation using a polar organic solvent such as methanol, ethanol, isopropanol, and acetone, electrodialysis, adsorption removal using an ion exchange resin, and membrane fractionation using a UF membrane. It is preferable to use these one method or a combination of two or more methods.
These purifications can be performed either before the demethoxy treatment described above or following the divalent metal and phosphate ion treatment described below. The refined or unpurified water-soluble soybean polysaccharide solution can be subjected to the subsequent treatment as it is or after being sterilized or dried.

(Bivalent metal ion and phosphate ion treatment)
The water-soluble polysaccharide having these acidic sugars such as water-soluble soybean polysaccharide is treated with a divalent metal ion and phosphate ions. The divalent metal ion is derived from an element classified as the second genus, preferably derived from calcium or magnesium, or a combination thereof, and more preferably derived from calcium. This divalent metal ion can be used in the form of various salts, preferably carbonates, chloride salts and hydroxide salts. Examples of phosphate ions include those derived from phosphoric acid, phytic acid, polyphosphoric acid, metaphosphoric acid or salts thereof, preferably those derived from phosphate ions having chelating ability, and more preferably phytic acid. Alternatively, it is derived from hexametaphosphoric acid or a salt thereof, most preferably derived from phytate.

  The divalent metal ion and phosphate ion treatment means that these substances are added directly to the aqueous solution of the water-soluble polysaccharide, or the divalent metal ion and / or phosphate ion is used as an aqueous solution, and the aqueous solution is used as the water-soluble polysaccharide. It is performed by adding to an aqueous solution. The addition amount of the divalent metal ion is preferably 1 to 300 mol%, more preferably 5 to 100 mol%, based on the acidic sugar residue of the water-soluble polysaccharide. If the amount of divalent metal ions is small, the effect of stabilizing the dispersion of protein particles, which will be described later, is difficult to obtain. The addition amount of phosphate ions is preferably 0.1 to 300 mol%, more preferably 1 to 30 mol%, based on the acidic sugar residue of the water-soluble polysaccharide. If there are few phosphate ions, it is difficult to obtain the effect of stabilizing the dispersion of protein particles, which will be described later.

(Two-stage processing)
The treatment with the divalent metal ions and phosphate ions for the water-soluble polysaccharide having an acidic sugar described above is performed first by treatment with divalent metal ions, followed by treatment with phosphate ions. In addition, the amount of metal ions and phosphate ions used can be reduced. Furthermore, if the aqueous solution of the water-soluble polysaccharide is purified by the various desalting methods described above after the treatment with the divalent metal ions, the subsequent treatment with the phosphate ions becomes more effective. When this two-stage treatment is performed, the amount of divalent metal ions added is preferably 1 to 100 mol%, more preferably 1 to 80 mol%, based on the acidic sugar residue of the water-soluble polysaccharide. The addition amount of phosphate ions is preferably 0.1 to 30 mol%, more preferably 0.1 to 15 mol%, and most preferably 0.5 to 5 mol% with respect to the acidic sugar residue of the water-soluble polysaccharide.

(Purification)
Phosphorylated water-soluble polysaccharides that have been treated with divalent metal ions and phosphates ions by simultaneous treatment or two-step treatment can be dried and used for various purposes, but they will exhibit more function. Therefore, it is desirable to perform the above-described purification such as desalting in order to remove excess divalent metal ions and phosphate ions added. The purified phosphorylated water-soluble polysaccharide solution is subjected to sterilization treatment such as plate sterilization or steam sterilization, followed by freeze drying, spray drying, heat drying, etc. Is obtained.

(Dispersion stabilizer)
The phosphorylated water-soluble polysaccharide of the present invention, particularly phosphorylated water-soluble soybean polysaccharide, is a protein dispersion stabilizer that can prevent aggregation of protein particles and maintain a stable dispersion state. The pH range is wide from about pH 3 to a pH range exceeding pH 4.2, preferably effective in the range of pH 3.4 to 4.6, more preferably pH 3.6 to 4.4. Suitable for beverages. However, if the milk protein is dispersed in a pH range of pH 4.2 or lower, it can be achieved with a conventional water-soluble soybean polysaccharide, and it may be difficult to achieve in the present invention in a pH range of less than 3.4. In addition, in the pH range exceeding pH 5.2, the milk protein starts to dissolve, and there is little need to use a dispersant. By using a dispersion stabilizer composed of phosphorylated water-soluble soybean polysaccharide produced according to the present invention, an acidic protein food or drink can be prepared in a pH range exceeding pH 4.2.

  Compared with conventionally used pectin and the like, the dispersion stabilizer of the present invention is easily sour and has a good flavor and can be used in a wide range of applications. There is also a feature of low viscosity. In order to improve the physical properties of the dispersion stabilizer, various gums and proteins and their degradation products can be used in combination as required. Examples of these combinations include starch, modified starch, various celluloses, agar, carrageenan, ferreran, guar gum, locust bean gum, tamarind seed polysaccharide, tara gum, gum arabic, tragacanth gum, karaya gum, pectin, xanthan gum, pullulan, gellan gum, etc. In addition to these polysaccharides, proteins such as gelatin can be exemplified.

(Acid protein food and drink)
The acidic protein food and drink in the present invention is an acidic food and drink containing protein derived from animals and plants, and beverages using animal and vegetable proteins such as milk and soy milk, or fruit juice, or organic acids such as citric acid and lactic acid, Gelatinization of acidic protein beverages made by adding inorganic acids such as phosphoric acid, acidic ice cream made by adding fruit juice to frozen desserts containing milk components such as ice cream, acidic frozen desserts such as frozen yogurt, pudding, bavalois, etc. It includes acidic desserts in which fruit juice is added to food, as well as protein foods and drinks with acidic properties such as coffee drinks, acidic creams, yogurt, lactic acid bacteria drinks (including bactericidal and live bacteria types), fermented milk, and kefir. Animal and vegetable proteins include cow milk, goat milk and other animal milk, soy milk, processed skim milk, whole milk powder, skim milk powder, whey powder, powdered soy milk, sweetened milk, condensed milk, concentrated milk, It refers to foods and drinks using processed milk and fermented milk that are enriched with minerals such as calcium and vitamins. A typical example of the acidic protein food and drink of the present invention is an acidic milk beverage in which milk protein is dispersed.

  The phosphorylated water-soluble soybean polysaccharide produced by the present invention exerts a function for stabilizing the dispersion of the protein, particularly in acidic protein beverages. Proteins can be dispersed and stabilized even in the pH range near the isoelectric point of proteins, which cannot be stabilized by conventional water-soluble soybean polysaccharides. As the protein species at that time, milk protein is most widely used as an acidic food and drink, and the effect of the dispersion stabilizer of the present invention is remarkably exhibited, which is preferable. The viscosity of the acidic protein beverage at that time varies depending on the concentration of the protein and the dispersion stabilizer. For example, in the formulation of Example 1 described later, the acidic protein having a viscosity of 10 mPa · s or less is used when the dispersion stabilizer of the present invention is used. Can be a beverage. In addition, lactic acid bacteria could not survive in conventional acidic milk beverages using pH 4.2 or less using water-soluble soy polysaccharides, whereas when the stabilizer of the present invention was used, foods and drinks in the pH range where lactic acid bacteria can survive were obtained. Therefore, an unheated live bacteria type lactic acid bacteria beverage using lactic acid bacteria fermented milk can be produced.

The protein dispersion stabilizer of the present invention is particularly effective in an acidic food or drink having a protein concentration of 10% by weight or less, preferably 0.05 to 2.0% by weight, more preferably 0.1 to 1.5% by weight, based on the acidic food or drink. More preferably, by adding 0.2 to 1.0% by weight, good dispersion stability of the protein is exhibited up to the pH range near the isoelectric point of the protein. For example, with milk proteins, protein foods and drinks having a pH of about 3.4 to 4.6 can be prepared, and aggregation can be particularly effectively suppressed at pH 3.6 to 4.4. When the dispersion stabilizer is at a high concentration, the flavor of the food or drink may be affected. At a low concentration, the dispersion stability may not be exhibited.

  Examples are described below. In the examples, “%” means a weight basis unless otherwise specified.

○ Production Example 1 (simultaneous mixing / hexametaphosphoric acid)
To 100 g of 10% aqueous solution of water-soluble soybean polysaccharide (Soya Five S-DA300: manufactured by Fuji Oil Co., Ltd.), add 13.78 g of sodium hexametaphosphate and 3.3 g of calcium chloride (both manufactured by Kishida Chemical Co., Ltd.) Stir at room temperature for 1 hour at pH 5.0. 100 g of 99 wt% ethanol was added, and the precipitate was recovered by centrifugation. The obtained precipitate was further washed sequentially with 70% by weight, 90% by weight and 95% by weight ethanol, and the precipitate was further dried on a hot plate to obtain phosphorylated water-soluble soybean polysaccharide B. Similarly, a solution containing 1.38 g of sodium hexametaphosphate and 0.33 g of calcium chloride was added to a solution of phosphorylated water-soluble soybean polysaccharide C, 0.14 g of sodium hexametaphosphate and 0.033 g of calcium chloride. Sugar D was designated. In addition, what refine | purified ethanol only about water-soluble soybean polysaccharide was prepared separately, and it was set as the water-soluble soybean polysaccharide A which is a comparative manufacture example.

○ Example 1 (Production and stability evaluation of acidic milk beverage)
(Preparation of fermented milk)
An aqueous solution containing 21% by weight of skim milk powder (manufactured by Yotsuba Milk Industry Co., Ltd.) was prepared and sterilized by heating at 95 ° C. with stirring. After cooling, commercially available plain yogurt was inoculated and fermented in a 40 ° C. incubator until the pH reached 4.6. The fermented yogurt was crushed and homogenized with a homogenizer (150 kgf) to obtain fermented milk.

(Preparation and evaluation of acidic milk beverages)
11.4 g of fermented milk (homogenized yogurt), 1.4 g of 50% sugar solution, 0.8% by weight of each aqueous solution of water-soluble soybean polysaccharide A or phosphorylated water-soluble soybean polysaccharides BD prepared in the above production examples 20 g was added to make the total weight 40 g (final concentration of water-soluble soybean polysaccharide 0.4% by weight). Further, the pH was arbitrarily adjusted with an aqueous solution of citric acid or sodium hydroxide. The treatment for 30 seconds was performed twice with an ultrasonic crusher (made by Kaijo Corp., 5281 type vibrator). This 1 ml was transferred to an Eppendorf tube, heated at 80 ° C. for 30 minutes, then cooled to 10-20 ° C., centrifuged at 300 rpm for 20 minutes, and the precipitation weight excluding the supernatant was measured to determine the precipitation rate. . The precipitation rate was calculated by the following formula.
Precipitation rate (%) = (precipitate weight) / (sorted acidic milk beverage weight) × 100
In the evaluation of the precipitation rate, ◎ indicates that the precipitation rate is 1% or less, ○ indicates that the precipitation rate exceeds 1% and 2% or less, △ indicates that the precipitation rate exceeds 2% and 4% or less, and X indicates that the precipitation rate exceeds 4%. did.

○ Table 1. Stability of acidic milk beverages with phosphorylated water-soluble soybean polysaccharide (simultaneous addition)

  Compared to the control soybean polysaccharide A, phosphorylated water-soluble soybean polysaccharides BD, to which calcium chloride and sodium hexametaphosphate were added, all had a reduced amount of precipitation when the acidic milk beverage was at pH 4.4 to 4.6. However, the dispersion of milk protein particles in this pH range was stabilized.

○ Production Example 2 (Two-stage treatment 1, hexametaphosphoric acid)
To 1,000 g of a 5 wt% aqueous solution of water-soluble soybean polysaccharide (Soya Five S-DA300), 3.68 g of calcium chloride was added, and the mixture was stirred at room temperature for 1 hour at pH 5.0. 1,000 g of 99 wt% ethanol was added, and the precipitate was recovered by centrifugation. The resulting precipitate was further washed sequentially with 70 wt% and 90 wt% ethanol and further air dried to obtain a Ca-treated water-soluble soybean polysaccharide.
0.4 g, 0.2 g, and 0.025 g of sodium hexametaphosphate were added to 100 g of a 5 wt% aqueous solution of Ca-treated water-soluble soybean polysaccharide, and stirred at room temperature for 1 hour at pH 5.0. 100 g of 99 wt% ethanol was added, and the precipitate was recovered by centrifugation. The obtained precipitate was further washed successively with 70% by weight and 90% by weight ethanol and further air-dried to obtain phosphorylated water-soluble soybean polysaccharides E to G.

○ Production Example 3 (Two-stage treatment 2, phytic acid)
In the same manner as in Production Example 2, phosphorylated water-soluble soybean polysaccharides H to J were obtained. However, instead of sodium hexametaphosphate, preparation was performed using 0.2 g, 0.1 g, and 0.05 g of sodium phytate (manufactured by sigma).

○ Example 2 (Production and stability evaluation of acidic milk beverage)
In the same manner as in Example 1, water-soluble soybean polysaccharide A and phosphorylated water-soluble soybean polysaccharides E to J were evaluated. In addition, each water-soluble soybean polysaccharide was evaluated by two kinds of systems having a final concentration of 0.4% by weight and 0.8% by weight with respect to an acidic milk beverage.

○ Table 2. Stability of acidic milk drinks with phosphorylated water-soluble soybean polysaccharide (two-stage addition) 0.4%

○ Table 3. Stability of acidic milk beverages with phosphorylated water-soluble soybean polysaccharide (two-stage addition) 0.8%

  In the system having a final concentration of 0.4% by weight (Table 2), the phosphorylated water-soluble soybean polysaccharide subjected to the treatment of the present invention had an overall reduced precipitation rate and improved milk protein particle dispersion stability. . In particular, phosphorylated water-soluble soybean polysaccharides F to J have a significantly reduced precipitation rate at pH 4.4, and it has been found that the dispersion stability in the vicinity of the isoelectric point of milk protein particles is improved. Further, in the system having a final concentration of 0.8% by weight (Table 3), the overall precipitation rate is further reduced, and in particular, in the phosphorylated water-soluble soybean polysaccharides H to J to which sodium phytate is added, the pH is 4.4. The precipitation rate was greatly improved.

○ Production Example 4 (application to pectin)
To 1,000 g of a 5 wt% aqueous solution of HM-pectin (CP kelco, YM-150-LJ), 3.68 g of calcium chloride was added and stirred at pH 5.0 at room temperature for 1 hour. 1,000 g of 99 wt% ethanol was added, and the precipitate was recovered by centrifugation. The resulting precipitate was further washed sequentially with 70% by weight and 90% by weight ethanol and further air-dried to obtain a Ca-treated pectin.
0.1 g of sodium phytate was added to 100 g of a 5% by weight aqueous solution of Ca-treated pectin, and the mixture was stirred at room temperature for 1 hour at pH 5.0. 100 g of 99 wt% ethanol was added, and the precipitate was recovered by centrifugation. The obtained precipitate was further washed successively with 70% by weight and 90% by weight ethanol and further air-dried to obtain phosphorylated pectin to which phytic acid was bound.

○ Production Example 5 (Application to carrageenan)
To 1,000 g of a 5 wt% aqueous solution of lambda carrageenan (manufactured by Saneigen FFI, carrageenin CSL-1), 0.032 g of magnesium chloride was added and stirred at pH 5.0 for 1 hour at room temperature. 1,000 g of 99 wt% ethanol was added, and the precipitate was recovered by centrifugation. The resulting precipitate was further washed sequentially with 70% by weight and 90% by weight ethanol and further air-dried to obtain Mg-treated carrageenan.
0.1 g of sodium phytate was added to 500 g of a 1% by weight aqueous solution of Mg-treated carrageenan, and the mixture was stirred at room temperature for 1 hour at pH 5.0. 500 g of 99 wt% ethanol was added, and the precipitate was recovered by centrifugation. The obtained precipitate was further washed successively with 70% by weight and 90% by weight ethanol and further air-dried to obtain a phosphorylated carrageenan bound with phytic acid.

Claims (9)

A method for producing a phosphorylated water-soluble soybean polysaccharide, comprising treating a water-soluble soybean polysaccharide having an acidic sugar as a constituent sugar with a divalent metal ion and a phosphate ion. To water-soluble soybean polysaccharide having an acidic sugar as a saccharide, and treated with divalent metal ions, followed characterized by treatment with phosphoric acid ions, phosphorylation water-soluble soybean polysaccharide of claim 1 Manufacturing method. The production method according to claim 1, wherein the divalent metal ion is derived from calcium or magnesium. The production method according to claim 1, wherein the phosphate ions are derived from one or more selected from phytic acid, metaphosphoric acid, and polyphosphoric acid. A phosphorylated water-soluble soybean polysaccharide produced by the method according to claim 1. A protein particle dispersion stabilizer comprising the phosphorylated water-soluble soybean polysaccharide according to claim 5 as an active ingredient. An acidic protein food containing the phosphorylated water-soluble soybean polysaccharide according to claim 5. A method for enhancing the ability to disperse protein particles of a water-soluble soybean polysaccharide, comprising treating the water-soluble soybean polysaccharide with a divalent metal ion and a phosphate ion. A method for enhancing the ability to disperse protein particles of a water-soluble soybean polysaccharide, characterized by treating the water-soluble soybean polysaccharide with a divalent metal ion and subsequently treating with a phosphate ion.