MX2012007892A - Stabilization of citrus fruit beverages comprising soy protein. - Google Patents

Abstract

A composition which permits protein fortification of citrus juices, particularly orange juice, or beverages containing citrus juices, to be carried out without separation of the juice or beverage and the rapid development of a clear or nearly clear liquid layer on top of the juice or beverage, comprises a soy protein product having a protein content of at least about 60 wt% (N x 6.25), preferably at least about 90 wt%, and preferably at least about 100 wt%, which is completely soluble in water at an acid pH value of less than about 4.4 and which is heat stable in aqueous solution, and at least one of at least one calcium salt and at least one organic acid.

Description

STABILIZATION OF CITRUS FRUIT BEVERAGES THAT COMPRISE SOY PROTEIN FIELD PE INVENTION This invention relates to the stabilization of citrus fruit drinks fortified with proteins.

BACKGROUND OF THE INVENTION In U.S. Patent Application No. 12/603, 087 filed October 21, 2009 (U.S. Patent Publication No. 2010-0098818, WO 2010/045727) (S701), assigned to the transferee of the and the descriptions of which are incorporated herein by reference, describes the production of a novel soy protein isolate that produces transparent and heat stable solutions at low pH values and, therefore, can be used to protein fortification, in particular, of carbonated beverages and sports drinks, as well as other aqueous systems, without precipitation of the protein.

The soy protein isolate provided herein has a unique combination of parameters not found in other soy isolates.

The product is completely soluble at acidic pH values of less than about 4.4 and the solutions thereof are stable to heat, allowing thermal processing, such as in hot fill applications. Stabilizers or other additives are not necessary to keep the protein in solution or suspension. The soy protein solution has no "bean" flavor and does not give off odors either. The product has low phytic acid content and enzymes are not required in the production of the soy protein isolate. The soy protein isolate is also highly soluble at about pH 7.

The novel soy protein isolate having a soy protein content of at least about 90% by weight (N x 6.25), preferably at least about 100% by weight, on a dry weight basis (db), It is produced by a method that comprises: (a) extract a source of soy protein with an aqueous solution of calcium salt, in particular calcium chloride solution, to cause solubilization of the soy protein from the protein source and form an aqueous solution of soy, (b) separating the aqueous solution of soy protein from the source of residual soy protein, (c) optionally dilute the aqueous solution of soy protein, (d) adjusting the pH of the aqueous solution of soy protein to a pH between about 1.5 and 4.4, preferably between about 2 and 4, to produce an acidified clear soy protein solution, (e) optionally heat treating the acidified solution to reduce the activity of the anti-nutritional trypsin inhibitors and the microbial load, (f) optionally concentrating the aqueous solution of clear soy protein while maintaining the ionic strength substantially constant by using a selective membrane technique, (g) optionally diafiltrate the concentrated soy protein solution, (h) optionally pasteurizing the concentrated soy protein solution to reduce the microbial load, and (i) optionally dehydrate the concentrated soy protein solution.

In an attempt to use the novel soy protein isolate for protein fortification of a variety of commercial orange juice products, the separation of orange juice components was observed along with the rapid development of a clear upper or liquid layer. almost clear (slightly cloudy) in the juice sample.

BRIEF DESCRIPTION OF THE INVENTION It has now been found that the novel soybean protein isolate can be used to provide citrus fruit juices fortified with proteins, without the rapid development of a clear or nearly clear upper liquid layer in the juice by using calcium salts alone, organic acids alone or the two species in combination.

Accordingly, in one aspect of the present invention, there is provided a composition comprising: a soy protein product having a protein content of at least about 60% by weight (N x 6.25) which is completely soluble in water at an acidic pH value of less than about 4.4 and which is heat stable in aqueous solution , and at least one of at least one calcium salt and at least one organic acid, the composition will be soluble in citrus fruit juices or drinks containing citrus fruit juices, without the separation of the components of the juice or citrus fruit drink and the rapid development of a substantially clear upper liquid layer in the juice or beverage.

As can be seen from the examples below, there are many possible formulations for the use of calcium salts and organic acids in the stabilization of orange juice and other juices and other mixed citrus fruit beverages containing these juices fortified by the isolated soy protein novel. Higher calcium levels are required to achieve stability when little or no organic acid is used, whereas lower calcium values can be used when higher values of organic acids are used.

In another aspect of the present invention, there is provided a juice or a citrus fruit drink fortified with proteins containing citrus fruit juice having dissolved therein the composition of the invention. The juice or citrus fruit drink fortified with proteins containing citrus fruit juice preferably has the composition: between about 0.1 and 10% w / w of soy protein from the soy protein product, and at least one between about 0 and 1.7% w / w of at least one calcium salt, and between about 0 and 1% w / w of at least one organic acid.

Suitable calcium salts include, but are not limited to, calcium chloride, calcium lactate, and calcium lactate gluconate. Suitable organic acids include, but are not limited to, malic acid and citric acid. Combinations of calcium salts and / or organic acids that have been found satisfactory to stabilize the orange juice against separation and the development of a substantially clear upper liquid layer when fortified with approximately 1.9% w / w protein isolate Soybeans include: - 1.04% w / w of calcium lactate alone - 0.08% w / w calcium lactate with 0.95% w / w malic acid - 0.08% w / w of calcium lactate with 0.95% w / w of citric acid - 0.75%) w / w of calcium lactate with 0.94% w / w malic acid - 0.76% w / w of calcium lactate with 0.47% w / w malic acid - 0.38% w / w of calcium chloride alone - 0.04 and 0.19% in w / w of calcium chloride and 0.95% in w / w of malic acid - 0.19% w / w calcium chloride with 0.48% w / w malic acid - 0.95 and 1.23% in w / w of calcium lactate gluconate alone - 0.09 and 0.47% in w / w of calcium lactate gluconate and 0.95% in w / w of malic acid 0. 48% w / w of calcium lactate gluconate and 0.48% w / w malic acid - 0.95% w / w malic acid alone It is evident that other combinations of calcium salts and / or organic acids will work in an equivalent manner.

While the present invention relates primarily to the use of soy protein isolates, it is contemplated that lower purity soy protein products with similar properties to soy protein isolate can be used. These lower purity products may have a protein concentration of at least about 60% by weight (N x 6.25) db GENERAL DESCRIPTION OF THE INVENTION The initial step of the process for providing the soy protein product used in the composition described herein involves solubilizing the soy protein from a source of soy protein. The source of soy protein can be soybeans or any soy product or by-product derived from the processing of soybeans., including, but not limited to, soybean meal, soy flakes, soybeans and soybean meal. The source of soy protein can be used in the form of total fat, in a partially defatted form or in a totally defatted form. When the source of soy protein contains an appreciable amount of fat, a step for oil removal is generally required during the process. The soy protein recovered from the soy protein source may be the protein that occurs in nature in soybeans or the protein material may be a protein modified by genetic manipulation, but possessing the characteristic hydrophobic and polar properties of the natural protein.

The solubilization of proteins from the soy protein source material is most conveniently carried out using a calcium chloride solution, although solutions of other calcium salts can be used. In addition, other alkaline earth metal compounds, such as magnesium salts, can be used. In addition, extraction of the soy protein from the soy protein source can be effected using calcium salt solution in combination with another saline solution, such as sodium chloride. Additionally, the extraction of the soy protein from the soy protein source can be carried out using water or another saline solution, such as sodium chloride, with calcium salt which will subsequently be added to the aqueous solution of soy protein. produced in the extraction step. The precipitate formed with the addition of the calcium salt is removed before further processing.

As the concentration of the calcium salt solution increases, the degree of solubilization of the protein from the soy protein source initially increases until a maximum value is reached. Any subsequent increase in salt concentration does not increase the total solubilized protein. The concentration of the calcium salt solution that causes the maximum solubilization of the protein varies depending on the salt being treated. In general, it is preferred to use a concentration value less than about 1.0 M, and more preferably a value between about 0.10 M and 0.15 M.

In a batch process, the saline solubilization of the protein is carried out at a temperature between about 1 ° C and 100 ° C, preferably between about 15 ° and 60 ° C, more preferably between about 15 ° C and about 35 ° C. ° C, preferably accompanied by stirring to decrease the time of solubilization, which is usually between about 1 and 60 minutes. It is preferred to carry out the solubilization to extract practically as much protein from the soy protein source as possible, in order to provide a high total yield of the product.

In a continuous process, the extraction of the soy protein from the soy protein source is carried out in any consistent manner by carrying out a continuous extraction of soy protein from the soy protein source. In one embodiment, the soy protein source is continuously mixed with a calcium salt solution and the mixture is transported through a tube or conduit having a length and a flow magnitude for a sufficient residence time to carry perform the desired extraction according to the parameters described herein. In this continuous process, the step of saline solubilization is carried out rapidly, in a time of up to about 10 minutes, preferably to carry out the solubilization to extract practically as much protein from the source of soy protein as possible. Solubilization in the continuous process is carried out at temperatures between about 1 ° C and 100 ° C, preferably between about 15 ° and 60 ° C, more preferably between about 15 ° C and 35 ° C.

The extraction is generally conducted at a pH between about 5 and 11, preferably between about 5 and 7. The pH of the extraction system (the source of soy protein and the calcium salt solution) can be adjusted to any value desired within the variation between about 5 and 11 to be used in the extraction step by the use of any convenient food grade acid, in general hydrochloric acid or phosphoric acid, or food grade alkali, in general sodium hydroxide, as necessary.

The concentration of the soy protein source in the calcium salt solution during the solubilization step can vary widely. Typical concentration values are between about 5 and 15% w / v.

The step for extracting proteins with the aqueous saline has the additional effect of solubilizing fats that may be present in the soy protein source, which then results in the fats being present in the aqueous phase.

The protein solution resulting from the extraction step generally has a protein concentration between about 5 and 50 g / L, preferably between about 10 and 50 g / L.

The aqueous calcium salt solution may contain an antioxidant. The antioxidant can be any convenient antioxidant, such as sodium sulfite or ascorbic acid. The amount of antioxidant employed can vary between about 0.01 and 1% by weight of the solution, preferably about 0.05% by weight. The antioxidant serves to inhibit the oxidation of any phenolic in the protein solution.

The aqueous phase resulting from the extraction step can then be separated from the residual soy protein source, in any convenient manner, such as by using a decanter centrifuge, followed by centrifugation and / or disk filtration, to remove the source material from residual soy protein. The source of separated residual soy protein can be dehydrated for disposal. Alternatively, the separated residual soy protein source can be re-extracted with freshly prepared calcium salt solution and the protein solution produced with the clarification combined with the initial protein solution for further processing as described below. Alternatively, the separate source of residual soy protein can be processed by a conventional isoelectric precipitation method or any other convenient method to recover this residual protein.

When the soy protein source contains significant amounts of fat, as described in U.S. Patent Nos. 5,844,086 and 6,005,076, assigned to the assignee thereof and the descriptions thereof are incorporated herein by reference, then the defatting steps described therein can be carried out in the separated aqueous protein solution. Alternatively, defatting of the separated aqueous protein solution can be achieved by any other convenient method.

The aqueous solution of soy protein can be treated with an absorbent, such as activated carbon powder or granulated activated carbon, to remove colored and / or odor compounds. This absorbent treatment can be carried out under any convenient conditions, generally at the ambient temperature of the aqueous solution of the separated pro tein. For activated carbon powder, an amount between about 0.025% and 5% w / v is used, preferably between about 0.05% and 2% w / v. The adsorbent agent can be removed from the soy solution by any suitable means, such as by filtration.

The aqueous solution of the resulting soy protein can be diluted in general with between about 0.5 and 10 volumes, preferably between about 0.5 and 2 volumes, of aqueous diluent. in order to decrease the conductivity of the aqueous solution of soy protein to a value usually below about 90 mS, preferably between about 4 and 18 mS. This dilution is usually carried out using water, although a diluted salt solution, such as sodium chloride or calcium chloride, having a conductivity of up to about 3 mS can be used.

The diluent with which the soy protein solution is mixed can have a temperature between about 2 ° and 70 ° C, preferably between about 10 ° and 50 ° C, more preferably between about 20 ° and 30 ° C.

The diluted soy protein solution is then adjusted in the pH to a value between about 1.5 and 4.4, preferably between about 2 and 4, by the addition of any suitable food grade acid, to result in an aqueous solution of acidified light soy protein. The aqueous acidified clear soy protein solution has a conductivity generally below about 95 mS, preferably between about 4 and 23 mS.

The aqueous solution of acidified light soy protein can be subjected to a heat treatment to inactivate thermolabile anti-nutritional factors, such as trypsin inhibitors, present in this solution as a result of extraction of the soy protein source material during the extraction step. This heating step also provides the additional benefit of reducing the microbial load. In general, the protein solution is heated to a temperature between about 70 ° and 160 ° C, for about 10 seconds and 60 minutes, preferably between about 80 ° and 120 ° C for about 10 seconds and 5 minutes, most preferably between approximately 85 ° and 95 ° C, for approximately 30 seconds and 5 minutes. The heat treated acidified soy protein solution can then be cooled for further processing as will be described below, at a temperature between about 2o and 60 ° C, preferably between about 20 ° and 35 ° C.

Optionally diluted protein solution, acidified and optionally heat treated, optionally can be clarified by any convenient means, such as by filtration, to remove any residual particles The resulting clear acidified soy protein aqueous solution can be dehydrated directly to prepare a soy protein product. In order to provide a soy protein product having a reduced content of impurities and a reduced salt content, such as a soy protein isolate, the aqueous solution of clear acidified soy protein can be processed prior to dehydration.

The aqueous solution of clear acidified soy protein can be concentrated to increase the protein concentration thereof while keeping the ionic strength of the same practically constant. This concentration is generally effected to provide a concentrated solution of soy protein having a protein concentration between about 50 and 300 g / L, preferably between about 100 and 200 g / L.

The concentration step can be carried out in any convenient manner consistent with a batch or continuous operation, such as by employing any convenient selective membrane technique, such as ultrafiltration or diafiltration, using membranes, such as hollow fiber membranes. or spiral membranes, with a suitable molecular weight cutoff, such as between about 3,000 and 1,000,000 Daltons, preferably between about 5,000 and 100,000 Daltons, having considered different materials and membrane configurations, and, for a continuous operation, sized to allow the desired degree of concentration as the aqueous solution of protein passes through the membranes.

As is well known, ultraf iltration and similar selective membrane techniques will allow low molecular weight species to pass through, while preventing higher molecular weight species from passing through. The low molecular weight species include not only the ionic species of the food grade salt but also the low molecular weight materials extracted from the source material, such as carbohydrates, pigments, low molecular weight proteins and anti-nutritional factors, such as trypsin inhibitors, which by themselves are low molecular weight proteins. The molecular weight cutoff of the membrane is usually selected to ensure retention of a significant proportion of the protein in the solution, while allowing contaminants to pass through having considered different materials and membrane configurations.

The concentrated soy protein solution can then be subjected to a diafiltration step using water or a dilute salt solution. The diafiltration solution may be at its natural pH or at a pH equal to that of the protein solution to be diafiltered or at any pH value therebetween. This diafiltration can be carried out using between approximately 2 and 40 volumes of diafiltration solution., preferably between about 5 and 25 volumes of diafiltration solution. In the diafiltration operation, additional amounts of contaminants are removed from the clear soybean protein aqueous solution by passing through the membrane with the permeate. This purifies the aqueous solution of clear protein and can also reduce its viscosity. The diafiltration operation can be carried out until there are no significant additional amounts of contaminants or visible color present in the permeate, or until the material retained has been sufficiently purified in such a way that, when dehydrated, it provides an isolate of Soy protein having a protein content of at least 90% by weight (N x 6.25) db This diaphragm can be carried out using the same membrane as for the concentration step. However, if desired, the diafiltration step can be effected using a separate membrane with different molecular weight cutoff, such as a membrane having a molecular weight cutoff in the range between about 3,000 and 1,000,000 Daltons, preferably between about 5,000 and 100,000 Daltons, having considered different membrane and configuration materials.

Alternatively, the diaphragm passage can be applied to the clear aqueous acidified protein solution before concentration or to the aqueous solution of partially concentrated clear acidified protein. Diafiltration can also be applied at multiple points during the concentration process. When the diafiltration is applied before the concentration or the partially concentrated solution, the resulting diafiltered solution can then be further concentrated. The viscosity reduction achieved by diafiltering multiple times as the protein solution is concentrated can allow a fully concentrated, higher final protein concentration to be reached. This reduces the volume of material that will dehydrate.

The concentration step and the diafiltration step can be effected herein such that the subsequently recovered soy protein product contains less than about 90% by weight of protein (N x 6.25) db, such as at least about 60% by weight of protein (N x 6.25) db By partially concentrating and / or partially diafiltering the aqueous solution of clear soy protein, it is possible to remove only partially the contaminants. This protein solution can then be dehydrated to provide a soy protein product with lower levels of purity. The soy protein product is still capable of producing clear protein solutions under acidic conditions.

In the diafiltration medium, an antioxidant may be present during at least part of the diafiltration step. The antioxidant can be any convenient antioxidant, such as sodium sulfite or ascorbic acid. The amount of antioxidant used in the diafiltration medium depends on the materials used and may vary between about 0.01 and 1% by weight, preferably about 0.05% by weight. The antioxidant serves to inhibit the oxidation of any phenolics present in the concentrated soy protein solution.

The concentration step and the optional diaphysis step can be carried out at any convenient temperature, generally between about 2 ° and 60 ° C, preferably between about 20 ° and 35 ° C, and over the period of time to carry out the desired degree of concentration and diafiltration. The temperature and other conditions used to some extent depend on the membrane equipment used to carry out the membrane processing, the desired protein concentration of the solution and the efficiency of contaminant removal for the permeate.

There are two major trypsin inhibitors in soybeans, namely the Kunitz inhibitor, which is a heat-labile molecule with a molecular weight of approximately 21,000 Daltons, and the Bowman-Birk inhibitor, a more heat-stable molecule with a molecular weight of 8,000. Daltons. The level of activity of the trypsin inhibitor in the final product of the soy protein can be controlled by manipulating the various process variables.

As noted above, the heat treatment of the aqueous solution of acidified light soy protein can be used to inactivate the thermolabile trypsin inhibitors. The partially concentrated or fully concentrated soy protein solution can also be heat treated to inactivate thermolabile trypsin inhibitors. When the heat treatment is applied to the acidified partially concentrated soy protein solution, the resulting heat treated solution can then be further concentrated.

In addition, the steps of concentration and / or diafiltration can be operated in a favorable manner for the removal of the trypsin inhibitors in the permeate along with the other contaminants. The removal of the trypsin inhibitors is stimulated by using a membrane of larger pore size, such as between about 30,000 and 1,000,000 Da, operating the membrane at elevated temperatures, such as between about 30 ° and 60 ° C, and by using volumes greater than the diafiltration medium, such as, for example, between about 20 and 40 volumes.

The acidification and membrane processing of the diluted protein solution at a lower pH between about 1.5 and 3 can reduce the activity of the trypsin inhibitor relative to the processing of the solution at a pH greater than about 3 and 4.4. When the protein solution is concentrated and diafiltered to the lower end of the pH variation, it may be convenient to increase the pH of the retained material before dehydration. The pH of the concentrated and diafiltered protein solution may be increased to the desired value, for example pH 3, by the addition of any convenient food grade alkali such as sodium hydroxide.

In addition, a reduction in the activity of the trypsin inhibitor can be achieved by exposing the soy materials to reducing agents that alter or rearrange the disulfide bonds of the inhibitors. Suitable reducing agents include sodium sulfite, cysteine and tracetele istein.

The addition of these reducing agents can be carried out in various stages of the general process. The reducing agent can be added with the soy protein source material in the extraction step, it can be added to the aqueous solution of clarified soy protein after the removal of the residual soy protein source material, it can be added to the solution Protein concentrate before or after diafiltration, or can be mixed dry with the dehydrated soy protein product. The addition of the reducing agent can be combined with a thermal treatment step and the membrane processing steps, as described above.

If it is desired to preserve the active trypsin inhibitors in the concentrated protein solution, this can be achieved by eliminating or reducing the intensity of the heat treatment step, without using reducing agents, by operating the concentration and diafiltration steps to the greater end of the process. pH variation, such as pH 3 to about 4.4, using a concentration and diaph iltration membrane with a smaller pore size, operating the membrane at lower temperatures and employing lower volumes of the diafil traction medium.

The concentrated and optionally diafiltered protein solution may be subjected to an additional defatting operation, if required, as described in U.S. Patent Nos. 5,844,086 and 6,005,076. Alternatively, defatting of the concentrated and optionally diafiltered protein solution can be achieved by any other convenient method.

The clear aqueous solution of concentrated and optionally diafiltered protein can be treated with an absorbent, such as activated carbon powder or granulated activated carbon, to remove colored and / or odor compounds. This absorbent treatment can be carried out under any convenient conditions, generally at the ambient temperature of the concentrated protein solution. For activated carbon powder, an amount between about 0.025% and 5% w / v is used, preferably between about 0.05% and 2% w / v. The adsorbent can be removed from the soy protein solution by any convenient means, such as by filtration.

The aqueous solution of concentrated acidified clear and optionally diafiltered soybean protein can be dehydrated by any convenient technique, such as spray drying or lyophilization. A pasteurization step can be carried out on the soy protein solution before dehydration. This pasteurization can be carried out under any suitable pasteurization conditions. In general, the concentrated and optionally diafiltered soy protein solution is heated to a temperature between about 55 ° and 70 ° C, preferably between about 60 ° and 65 ° C, for between about 30 seconds and 60 minutes, preferably between approximately 10 and 15 minutes. The pasteurized concentrated soy protein solution can then be cooled for dehydration, preferably at a temperature between about 25 ° and 40 ° C.

The dehydrated soy protein product has an excess protein content of about 60% by weight (N x 6.25) d.b. Preferably, the dehydrated soy protein product is an isolate with a high protein content, in excess of about 90% by weight of protein, preferably at least about 100% by weight (N x 6.25) d.b.

As mentioned above, in an attempt to use this soy protein isolate for protein fortification of a variety of commercial orange juice products, the separation of components and the development of a substantially clear upper liquid layer in the juice were observed. of Orange. According to the present invention, the calcium salts, organic acids or the two species in combination can be used to allow the isolate of soy protein that is used to provide the citrus fruit juices fortified with proteins without the rapid development of a clear or almost transparent upper liquid layer in fruit juice, in particular orange juice. Higher calcium levels are required to achieve stability when little or no organic acid is used, whereas lower calcium values can be used where higher values of organic acid are used.

EXAMPLES Example 1: This example illustrates the production of soy protein isolate soluble in novel acid. 'a' kg of soybean meal minimally processed with heat, defatted was added to > b 'L of a 0.15M NaCl2 solution at room temperature and stirred for 60 minutes to provide an aqueous solution of protein. The residual soybean meal was removed and the resulting protein solution was clarified by centrifugation and filtration to produce 'c' L of the filtered protein solution having a protein content of xd '% by weight.

The filtered protein solution was then added to * e 'volumes of purified water by reverse osmosis and the pH of the sample was decreased to' f 'with diluted HC1.

The solution of the diluted and acidified protein extract was reduced in volume from ¾g 'L to * h' L by concentration in a membrane 'i' having a molecular weight cutoff of * j 'Daltons. The acidified concentrated protein solution was diafiltered with 'k' L purified water by reverse osmosis. The resultant, acidified, diafiltered concentrated protein solution had a protein content of 1% by weight and represented a yield of m% by weight of the initial filtered protein solution. 'n' kg of acidified, diafiltered, concentrated protein solution were passed through o 'L bed volumes of granular activated carbon at a flow magnitude of'? ' bed volumes per hour and then dehydrated to provide a product that was found to have a protein content of 4q '% (N x 6.25) d.b. The product was given the designation '' S701C.

In the following Table 1 the parameters' a 'to ¾r' of three executions are shown. S701C of the three runs was combined dry in the proportion of 46.6% by weight of S005-K18-08A S701C: 40.7% by weight of S005-K24-08A S701C: 12.7% by weight of S005-L08-08A S701C for form a product called Mixture I of S701C.

TABLE 1 - PARAMETERS FOR EXECUTIONS TO PRODUCE S701C FOR THE MIX I OF S701C N / A = not available Example 2: This example illustrates the production of another batch of soy protein isolate soluble in novel acid. 98. 34 kg of defatted soybean meal, minimally processed with heat, were added to 1,000 L of 0.15 M CaCl2 solution at room temperature and stirred for 30 minutes to provide an aqueous solution of protein. The residual soybean meal was removed and the resulting protein solution was clarified by centrifugation and filtration to yield 670.1 L of filtered protein solution having a protein content of 2.38% by weight.

The filtered protein solution was then added to 1 volume of purified water by reverse osmosis and the pH of the sample was decreased to 3.14 with diluted HC1.

The solution of the diluted and acidified protein extract was reduced in volume from 1,350 L to 100 L by concentration in a polyethersulfone membrane (PES) having a molecular weight cutoff of 100,000 Daltons. The concentrated, acidified protein solution was diafiltered with 1,000 L of purified water by reverse osmosis. The resulting acidified, diafiltered, concentrated protein solution had a protein content of 8.95% by weight and represented a yield of 74.6% by weight of the initial filtered protein solution. The concentrated, acidified, diafiltered protein solution was then dried to provide a product that was found to have a protein content of 101.31% (N x 6.25) d.b. The product was given the designation S008-C02-09A S701.

In case 3; This example illustrates the effect of the addition of the novel soy protein isolate to commercial orange juice products.

Sufficient isolate of powdered soy protein (S701C) from batch SO 05-K24-08A, prepared as described in Example 1, was added to commercial orange juice products to provide a protein concentration of 2% p / vy was solubilized with a magnetic stirrer. The protein fortified products were stored at 4 ° C for 24 hours and visually observed after 1 and 24 hours. The commercial orange juice products tested were Tropicana Essentials Low Acid Orange Juice, Tropicana Essentials Calcium Orange Juice, Tropicana Essentials Omega-3 Orange Juice, Tropicana Premium No Pulp Orange Juice and Tropicana Premium Orange Juice with Pulp.

After storage for one hour at 4 ° C, some sedimentation of solids was observed in all samples of orange juice except for the product of Tropicana Essentials Calcium Orange Juice, which appeared to be homogeneous without any separation. After 24 hours of storage at 4 ° C, all samples had separated with the development of a clear or almost clear upper liquid layer (referred to herein as clarification).

E xemployment 4: This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate of the present using malic acid.

In glass vials the powdered soy protein, prepared as described in Example 2, malic acid and Sun-Rype Orange Juice (aseptically processed) was weighed according to the formulations shown in Table 2.

TABLE 2 - FORMULATIONS FOR MASS ACID TRIALS Sample number 1 2 ugo of naran a by weight (g) 30.65 30.65 Protein powder by weight (g) 0.63 0.63 malic acid by weight (g) 0.15 0.30 % protein (w / w) 1.91 1.90 % malic acid (w / w) 0.48 0.95 The vials were mixed with a vortex mixer operated at medium speed until the aggregate compounds were completely dissolved. A sample of control orange juice was emptied into a glass vial without malic acid and without soy protein. The samples were placed in storage at 4 ° C and visually observed after 24 hours. After storage for 24 hours at 4 ° C, 0.48% w / w of the malic acid sample had separation with clarification. After the same storage period, the sample that contained 0.95% w / w malic acid did not show separation with clarification.

Apparently, malic acid alone was able to stabilize the Sun-Rype Orange Juice which contained approximately 1.9% w / w of the novel soy protein when used at a level of 0.95% w / w.

Example 5; This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate therein using calcium lactate.

The soy protein powder, prepared as described in Example 1, calcium lactate and Sun-Rype Orange Juice (aseptically processed) was weighed into glass vials in accordance with the formulations shown in Table 3.

TABLE 3 - FORMULATIONS FOR TESTS WITH CALCIUM LACTATE Sample number orange juice by weight (g) 30 .65 30 .65 30 .65 30 .65 protein powder by weight (g) 0. .62 0. 62 0. 62 0. 62 calcium lactate by weight (g) 0. 024 0. 12 0. 24 0. 33 % protein (w / w) 1,, 91 1. 91 1. 90 1. 89 % calcium lactate (w / w) 0. .08 0. 38 0. 76 1. 04 The vials were mixed with a vortex mixer operated at medium speed until the aggregate compounds were completely dissolved. A sample of control orange juice was emptied into a glass vial without soy protein and calcium lactate. The samples were placed in storage at 4 ° C and visually observed after 24 hours.

In the following Table 4 the obtained results are shown: TABLE 4 - OBSERVATIONS OF THE SÜN-RYPE ORANGE JUICE CONTAINING PROTEIN OF SOYA AND CALCICO LACTATE % calcium lactate (w / w) Observation 0. 08 Separation with clarification 0. 38 Separation with clarification 0. 76 Separation with clarification 1. 04 same appearance as the control orange juice sample As can be seen from the results presented in Table 4, the samples of orange juice that contained approximately 1.9% w / w protein and 0.08%, 0.38% and 0.76% w / w calcium lactate were not stable or showed separation with clarification. The sample that contained 1. 04% in p / p of calcium lactate, however, did not have separation with clarification nor had an appearance similar to the control sample.

Example 6: This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate therein using calcium lactate and malic acid.

The soy protein powder, prepared as described in Example 1, calcium lactate, malic acid and Sun-Rype Orange Juice (aseptically processed) according to the formulations shown in Table 5 was weighed into glass vials.

TABLE 5 - FORMULATIONS FOR TESTS WITH CALCIUM AND ACID LACTATE MALIC Sample number 1 2 3 4 orange juice by weight (g) 30 .65 30 .65 30 .65 30 .65 protein powder by weight (g) 0 .62 0 .62 0 .62 0 .62 calcium lactate in weight (g) 0. 024 0. 024 0 -.24 0. .24 malic acid by weight (g) 0 .03 0 .30 0, .03 0, .30 % protein (w / w) 1.91 1.89 1.90 1.88% calcium lactate (w / w) 0 .08 0. .08 0 .76 0. .75% malic acid (w / w) 0 .10 0.. 95 0 .10 0. .94 The vials were mixed with a vortex mixer operated at medium speed until the aggregate compounds were completely dissolved. A sample of control orange juice was emptied into a glass vial without soy protein, calcium lactate or malic acid. The samples were stored at 4 ° C and visually observed after 24 hours.

In the following Table 6 the obtained results are shown: TABLE 6 - OBSERVATIONS OF THE SÜN-RYPE ORANGE JÜICE CONTAINING PROTEIN OF SOYA, LACTATQ CALCICO AND ACIDO MÁLICO As can be seen from the results in Table 6, the samples containing 0.1% w / w malic acid exhibited separation with clarification, while the samples with the highest levels of malic acid did not have separation with clarification and They seemed similar to the control sample.

And 7; This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate therein using calcium chloride.

The procedure of Example 5 was repeated with the substitution of calcium lactate for calcium chloride. The formulations used are shown in the following Table 7.

TABLE 7 - FORMULATIONS FOR TESTS WITH CALCIUM CHLORIDE Sample number 1 2 3 4 orange juice by weight (g) 30.65 30.65 30 .65 30.65 protein powder by weight (g) 0.62 0.62 0.62 0.62 calcium chloride by weight (g) 0.012 0.03 0, .06 0.12 % protein (w / w) 1.91 1.91 1, .91 1.91 % calcium chloride (w / w) 0.04 0.10 0, .19 0.38 In the following Table 8 the obtained results are shown: TABLE 8 OBSERVATIONS OF THE SUN-RYPE ORANGE JUICE CONTAINING SOY PROTEIN AND CALCIUM CHLORIDE % calcium chloride (w / w) Observation 0. 04 Separation with clarification 0. 10 Separation with clarification 0. 19 Separation with clarification 0. 38 same appearance as the control orange juice sample As can be seen from the results provided in Table 8, the samples containing 0.04%, 0.10% and 0.19% w / w of calcium chloride were unstable and exhibited separation with clarification. However, the sample with 0.38% w / w of calcium chloride had no separation with clarification and seemed similar to the control sample.

Example 8; This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate therein using calcium chloride and malic acid.

The procedure of Example 6 was repeated substituting calcium lactate for calcium chloride. The formulations used are shown in the following Table 9.

TABLE 9 - FORMULATIONS FOR TESTS WITH CALCIUM CHLORIDE AND MALIC ACID Sample number 1 2 3 4 orange juice by weight (g) 30 .65 30 .65 30 .65 30 .65 protein powder by weight (g) 0, .62 0, .62 0. 62 0. 62 chloride of calcium by weight (g) 0. 012 0. 012 0. 06 0. 06 malic acid by weight (g) 0. .03 0. .30 0. 03 0. 30 % protein (w / w) 1. .91 1. .89 1. 91 1. 89 % calcium chloride (w / w) 0. .04 0, .04 0. 19 0. 19 % malic acid (w / w) 0 .10 0 .95 0. 10 0. 95 In the following Table 10 the obtained results are shown: TABLE 10 OBSERVATIONS OF THE SÜN-RYPE ORANGE JÜICE QDE CONTAINS PROTEIN OF SOYA, CLQRÜRO OF CALCIUM AND ACID MÁLICQ % acid chloride% Observation calcium (w / w) malic (w / w) 0. .04 0. .10 Separation with clarification 0. .04 0. .95 Same aspect as the control orange juice sample 0. .19 0. .10 Separation with clarification 0. .19 0. .95 same look as the orange juice control sample As can be seen from the results presented in Table 10, samples containing 0.1% w / w malic acid exhibited separation with clarification, while samples with 0.95% w / w malic acid appeared similar to the control sample.

Example 9: This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate therein using calcium lactate gluconate.

The procedure of Example 5 was repeated with calcium lactate gluconate (CLG) in substitution of calcium lactate. The formulations used are shown in the following Table 11.

TABLE 11 - FORMULATIONS FOR TESTS WITH GLUCONATE DE CALCICO LACTATE Sample number 1 2 3 4 juqo orange by weight (g) 35 .75 35.75 35.75 35.75 protein powder by weight (g) 0 .73 0.73 0.73 0.73 calcium lactate gluconate by weight (g) 0. 035 0.175 0.350 0.455 % protein (w / w) 1 .93 1.92 1.91 1.91 % of calcium lactate gluconate (w / w) 0 .10 0.48 0.95 1.23 In the following Table 12 the results obtained are shown: TABLE 12 - OBSERVATIONS OF THE SUN-RYPE ORANGE JUICE CONTAINING PROTEIN OF SOYA AND GLUCONATE OF LACTATE CALCICO As can be seen from the results presented in Table 12, samples of orange juice containing 0.10% and 0.48% w / w calcium gluconate gluconate were not stable or exhibited separation with clarification. The samples with 0.95% and 1.23% w / w calcium gluconate gluconate had no separation with clarification nor did they appear similar to the control sample.

Example 10: This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate therein using calcium lactate gluconate and malic acid.

The procedure of Example 6 was repeated, replacing calcium lactate with calcium lactate gluconate. In the following Table 13 the formulations used are shown.

TABLE 13 - FORMULATIONS FOR TESTS WITH GLUCONATE OF CALCIUM LACTATE AND MALIC ACID Sample number 1 2 3 4 orange juice by weight (g) 30 .65 30 .65 30 .65 30 .65 protein powder by weight (g) 0, .62 0 .62 0 .62 0, .62 gluconate of calcium lactate by weight (g) 0, .03 0 .30 0 .03 0, .30 malic acid by weight (g) 0 .03 0 .30 0 .03 0 .30 % protein (w / w) 1 .91 1 .89 1 .90 1 .89 % of calcium lactate gluconate (w / w) 0 .10 0 .09 0 .48 0 .47 % malic acid (w / w) 0 .10 0 .95 0 .10 0 .95 In the following Table 14 the obtained results are shown: TABLE 14 - OBSERVATIONS OF THE SÜN-RYPE ORANGE JÜICE CONTAINING PROTEIN OF SOY, CLG AND MALIC ACID % acid gluconate% Observation malic calcium lactate (w / w) (P / P) 0. .10 0., 10 Separation with clarification 0, .09 0. .95 Same aspect as the control orange juice sample 0. .48 0. .10 Solids more settled than the control orange juice sample, but without separation with clarification 0, .47 0, .95 Same aspect as the control orange juice sample As can be seen from the results presented in Table 14, the samples that contained 0.95% w / w malic acid were more stable than the samples that contained 0.10% w / w malic acid and appeared similar to the shows control. The sample with 0.48% w / w of CLG and 0.1% w / w of malic acid appeared to contain more settled solids than the control orange juice sample, but had an opaque top layer, while separation with clarification was observed for the sample with 0.1% w / w of CLG and 0.1% w / w of malic acid.

Example 11; This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate therein using malic acid together with calcium lactate, calcium chloride or calcium lactate gluconate.

The soy protein powder, prepared as described in Example 1, the calcium salts, the malic acid and dom-Rype Orange Juice (processed aseptically) were weighed in glass vials according to the formulations shown in Table 15 .

TABLE 15 - FORMULATIONS FOR TESTS WITH ISOLATION OF SOY PROTEIN OF EXAMPLE 1 Sample number 1 2 3 orange juice by weight (g) 30 .65 30 .65 30 .65 protein powder by weight (g) 0, .62 0, .62 0 .62 calcium lactate by weight (g) 0 , .24 0, .00 0 .00 calcium chloride by weight (g) 0. .00 0. .06 0 .00 calcium lactate gluconate by weight (g) 0 .00 0 .00 0 .15 malic acid in weight (g) 0. .15 0 .15 0 .15 % protein (w / w) 1. .89 1 .90 1 .89 % calcium lactate (w / w) 0 .76 0 .00 0 .00 % calcium chloride (w / w) 0 .00 0 .19 0 .00 % of calcium lactate gluconate (P / P) 0 .00 0 .00 0 .48 % malic acid (w / w) 0 .47 0 .48 0 .48 The samples were also prepared with the soy protein powder, prepared as described in Example 2, the calcium salts, malic acid and Sun-Rype Orange Juice (aseptically processed) weighed in glass vials in accordance with the formulations shown. in Table 16.

TABLE 16 - FORMULATIONS FOR TESTS WITH ISOLATION OF SOY PROTEIN OF EXAMPLE 2 Sample number 1 2 3 orange juice by weight (g) 30 .65 30 .65 30 .65 protein powder by weight (g) 0 .63 0 .63 0. .63 calcium lactate by weight (g) 0. 024 0 .00 0. .00 calcium chloride by weight (g) 0 .00 0. 012 0 .00 calcium lactate gluconate by weight (g) 0 .00 0 .00 0 .03 malic acid by weight (g) 0 .15 0 .15 0 .15 % protein (w / w) 1 .91 1 .91 1 .91 % calcium lactate (w / w) 0 .08 0 .00 0 .00 % calcium chloride (w / w) 0 .00 0 .04 0 .00 % calcium lactate gluconate (w / w) 0 .00 0 .00 0 .10 % malic acid (w / w) 0 .48 0 .48 0 .48 The samples were mixed with a vortex mixer operated at medium speed until the aggregate compounds were completely solubilized. The samples were placed in storage at 4 ° C and visually observed after 24 hours. A control sample was prepared without the soy protein, malic acid or calcium salt present.

The following results are shown in the following Tables 17 to 19: TABLE 17 - OBSERVATIONS OF THE SÜN- YPE ORANGE JÜICE CONTAINING PROTEIN OF SOYA, LACTATQ CALCICO AND ACIDO MALICQ % lactate% acid Observation calcium (p / p) malic (p / p) 0. 08 0.48 Separation with clarification 0. 76 0.47 Same aspect as the control orange juice sample TABLE 18 - OBSERVATIONS OF SÜN-RYPE ORANGE JUICE CONTAINING SOY PROTEIN, CALCIUM CHLORIDE AND MALICQ ACID % chloride acid% Observation calcium (w / w) malic (w / w) 0. 04 0.48 Separation with clarification 0. 19 0.48 Same aspect as the control orange juice sample TABLE 19 - OBSERVATIONS OF THE SUN-RYPE ORANGE JUICE CONTAINING PROTEIN OF SOYA, GLUCONATE OF CALCIUM LACTATE AND MALIC ACID % gluconate acid% Observation of malic lactate (w / w) calcium (p / p) 0. 10 0.48 Separation with clarification 0. 48 0.48 Same aspect as the control orange juice sample As can be seen from the results presented in Tables 17 to 19, the samples with lower levels of calcium showed separation with clarification while those with higher levels of calcium did not have separation with clarification and seemed similar to the control sample.

Example 12; This example illustrates the thermal stability of an orange juice product that contained the novel soy protein isolate and various amounts of calcium and malic acid salts.

The soy protein powder, prepared as described in Example 2, the calcium salts, the malic acid and the sun-Rype Orange Juice (processed aseptically) were weighed in beakers according to the formulations shown in the Table. twenty.

TABLE 20 - FORMULATIONS FOR THE THERMAL TREATMENT TEST Sample number 1 2 3 4 5 6 7 juqo orange at 204.30 204.30 204.30 204.30 204.30 204.30 204.3C weight (q) protein powder in 4 .19 4 .19 4 .19 4 .19 4 .19 4 .19 4.19 weight (q) calcium lactate in 0 .00 0 .16 0 .00 0 .00 2 .20 0 .00 0.00 weight (q) calcium chloride in 0 .00 0 .00 0 .08 0 .00 0 .00 0 .80 0.00 weight (g) CLG by weight (q) 0 .00 0 .00 0 .00 0 .20 0 .00 0 .00 2.00 malic acid by weight 0 .00 2 .00 2 .00 2 .00 0 .00 0 .00 0.00 l £ i % protein (w / w) 1 .92 1 .90 1 .90 1 .90 1 .90 1 .91 1.90 % calcium lactate 0 .00 0 .08 0 .00 0 .00 1 .04 0 .00 0.00 (p / p) ! of calcium chloride 0 .00 0 .00 0 .04 0 .00 0 .00 0 .38 0.00 (P / P) % of CLG (p / p) 0.00 0.00 0.00 0.09 0.00 0.00 0.95 Malic acid% 0.00 0.95 0.95 0.95 0.00 0.00 0.00 (P / P) The mixtures were stirred with a magnetic stirrer for one hour. The resulting samples they were treated with heat at 85 ° C for 30 seconds and then cooled in a bath with ice. The samples were transferred to food-grade plastic bottles, placed in storage at 4 ° C and visually observed after 24 hours.

In the following Table 21 the obtained results are shown: TABLE 21 - OBSERVATIONS OF SUN-RYPE ORANGE JUICE TREATED WITH HEAT CONTAINING SOY PROTEIN AND CALCIUM SALTS WITH OR WITHOUT MALIC ACID Calcium concentration% acid Observation added (% in p / p) malic (p / p) 0. 00 0. 00 Separation with clarification 0. 08% lactate 0. 95 No separation with Calcification clarification 1. 04% lactate 0. 00 Without separation with Calcification clarification 0. 04% chloride of 0. 95 Without separation with calcium clarification 0. 38% chloride of 0. 00 Without separation with calcium clarification 0. 09% of CLG 0. 95 Without separation with clarification 0. 95% of CLG 0. 00 Without separation with clarification As can be seen from the results provided in Table 21, the sample that contained the soy protein without malic acid or calcium salt showed separation with clarification.

The remaining samples did not have separation with clarification.

It can be concluded from these data that the stability of sun-Rype Orange Juice which contained the novel soy protein isolate stabilized with malic acid and the calcium salt was not adversely affected by the thermal treatment at 85 ° C.

Example 13 t This example illustrates attempts to stabilize an orange juice product having the novel soy protein isolate therein using calcium lactate and citric acid.

The soy protein powder, prepared as described in Example 1, calcium lactate, citric acid and Sun-Rype Orange Juice (processed aseptically) were weighed in glass vials according to the formulations shown in Table 22.

TABLE 22 - FORMULATIONS FOR TESTS WITH CALCIUM LACTATE AND CITRIC ACID Sample number Naran juice by weight (g) 20 .43 20 .43 protein powder by weight (g) 0. .41 0. 41 calcium lactate by weight (g) 0. 016 0. 016 citric acid by weight (g) 0. .00 0 .2 % protein (w / w) 1. .90 1. 88 % calcium lactate (w / w) 0. .08 0. 08 % citric acid (w / w) 0. .00 0. 95 The vials were mixed with a vortex mixer operated at medium speed until the aggregate compounds were completely dissolved. A sample of control orange juice was emptied into a glass vial without soy protein, calcium lactate or citric acid. The samples were stored at 4 ° C and visually observed after 24 hours.

In the following Table 23 the obtained results are shown: TABLE 23 - OBSERVATIONS OF SÜN-RYPE ORANGE JUICE CONTAINING PROTEIN OF SOY. CALCIUM LACTATE AND CITRIC ACID As can be seen from the results provided in Table 23, the sample that contained the soy protein and 0.08% w / w of calcium lactate alone, showed separation with clarification, while the sample with the same lactate level Calcium plus 0.95% w / w of citric acid did not have separation with clarification and seemed similar to the control sample.

SUMMARY OF THE DESCRIPTION In the summary of this description, the unstable solutions of citrus fruits fortified with the soy protein isolate can be stabilized against the separation of citrus fruit components and the rapid development of a clear or almost clear upper liquid layer by the use of calcium salts, organic acids or the two species in combination. Modifications are possible within the scope of this invention.

Claims (10)

1. A composition characterized by: a soy protein product having a protein content of at least 60% by weight (N x 6.25) which is completely soluble in water at an acidic pH value of less than 4.4 and which is stable to heat in aqueous solution, and at least one of at least one calcium salt and at least one organic acid, the composition will be soluble in citrus fruit juices or drinks containing citrus fruit juices, without the separation of the components of the juice or citrus fruit drink and the rapid development of a substantially clear upper liquid layer in the juice or beverage.

2. The composition according to claim 1, characterized in that at least one calcium salt is selected from the group consisting of calcium chloride, calcium lactate and calcium lactate gluconate.

3. The composition according to claim 1 or 2, characterized in that at least one organic acid is malic acid or citric acid.

4. The composition according to any of claims 1 to 3, characterized in that the juice of citrus fruits is orange juice.

5. The composition according to any of claims 1 to 4, characterized in that the soy protein product has a protein content of at least 90% by weight (N x 6.25) d.b.

6. The composition according to claim 5, characterized in that the soy protein product has a protein content of at least 100% by weight (N x 6.25) d.b.

7. A juice or citrus fruit drink fortified with proteins containing fruit juice citrus fruits having dissolved therein the composition according to claims 1 to 6.

8. The juice or citrus fruit drink containing citrus fruit juice according to claim 7, which is orange juice fortified with proteins.

9. The juice or citrus fruit drink containing citrus fruit juice according to claim 7 or 8, the composition is characterized by: 0. 1 to 10% w / w of soy protein from a soy protein product, and at least one of 0 and 1.7% w / w of at least one calcium salt, and 0 to 1% w / w of at least one organic acid.

10. The juice or drink of citrus fruits containing the juice of citrus fruits according to claim 9, characterized in that at least one calcium salt is selected from the group consisting of calcium chloride, calcium lactate and calcium lactate gluconate and at least one acid Organic is one of malic acid and citric acid.