MXPA99010313A - Method for treating a lactic raw material containing gmp - Google Patents

Abstract

The invention concerns a simple ion-exchanging industrial process which consists in treating a liquid lactic raw material containing glycomacropeptide in the presence of a weak anionic resin to obtain an improved protein product useful in food processing and said glycomacropeptide which is selectively adsorbed on the resin, then eluted in said resin. Before being treated with resin, the liquid raw material is decationized so that its pH has a value between 1 and 4.5.

Description

PROCESS TO TREAT A LACTICAL RAW MATERIAL THAT HAS GMP DESCRIPTION OF THE INVENTION - The invention relates to a process - for the treatment of a lactic raw material containing g 1 ic or cro epiphion or cation or cro-peptide (hereinafter GMP), for the purpose of to separate said GMP from it. - GMP is a partially silylated, oriferous opiate, which is formed by the action of a protease, for example, rennet, in kappa-casein of mammalian milk. Approximately 20% by weight of the proteins are present in the sweet whey obtained after separation of casein during the manufacture of cheese. A process for the manufacture of GMP at the laboratory level is known, which consists of treating a raw material of lactic origin, "such as, for example, an acid casein or a caseinate, which can be hydrolyzed through rennet, or alternately a lamellar-free, demineralized sweet whey from cheese making, with trichloroacetic acid in order to precipitate the proteins, and then recover the supernatant, dialyze, and finally dry the dialysate.This process is not industrial. Production of GMP on an industrial scale, which is described in EP-A-0,488,589, consists in treating a whey product through ion exchange, in order to recover the fraction that has not been adsorbed, concentrate it and demineralize it through ultrafiltration, diafiltration and, when reverse osmosis is appropriate and to recover GMP. A process for the reduction of a whey protein fraction of milk is described in OK-A-2, 188,526. It consists in treating a milk product with a strong anionic resin, under conditions so that the proteins and some peptides of the material are non-selectively adsorbed on the resin in the form of complexes. Such complexes are difficult for subsequently be eluted from the resin. The eluted product is characterized by the formation of a firm gel at a pH of less than 4.5 and at room temperature once it is suspended in water. The protein fraction can be used in stirred milk type drinks and in dessert creams.

In JP-A-O 7132 O 49, it is proposed to use a weakly anionic ion exchange resin, whose matrix is hydrophilic to separate the silylated peptides from the whey. The method used consists of passing the starting material, whose pH has been precisely adjusted in advance to a value of 4 to 6, on a hydrophilic macromolecular support consisting of a natural polysaccharide or a synthetic polyvinyl, inserted with basic exchange groups. The supports used as a matrix are not easily applicable in industrial form. The object of the invention is the selective separation of GMP from other components - from plastic materials in a single operation, to an industrial scale, with a high yield. Therefore, the invention relates to a process for the ion exchange treatment of a liquid lactic raw material --- consisting of GMP, with the help of recovering, on the one hand, a product that can be used directly as a source of protein and, on the other hand, the GMP in a "purified" form, characterized in that it comprises the following steps: i) of s ion t ioni zation of liquid liquid material, so that the pH has a value of 1 to 4.5, ii) bring the liquid into contact with an anionic resin of hydrophobic matrix, predominantly in the form "alkaline to a stabilized pH, iii) separation of the resin and the liquid product that is recovered, and iv) desorption of the GMP of the resin. As the lactic starting material, any product or sub-product containing GMP can be used in the process according to the invention. The following can be mentioned as a guide: "~ - s ~ ú ~ e r" of sweet milk obtained after the separation of casein coagulated with rennet, - sweet whey - or smi nei milk whey to a greater or lesser degree, for example, through electrodialysis, ion exchange, reverse osmosis, electrodeionization or a combination of these procedures, - a sweet milk concentrate, - a concentrate of sweet demineralized whey to a greater or lesser degree, for example, through electrodialysis, ion exchange, reverse osmosis, electrodeionization or a combination of these procedures, a concentrate of Sweet milk dream proteins substantially lactose-free, obtained from, for example, ultrafiltration, followed by diafiltration (ultrafiltration with washing), mother liquors of crystallization of lactose from sweet whey, a permeate of ultrafiltration of a sweet whey, - the hydrolysis product, through a protease, of a native casein obtained through the preparation of acid It is skimmed with an inorganic acid or through biological acidification, which when appropriate with addition of calcium ions or alternatively of a micellar casein, obtained for example, through my cr of a skim milk, - the product of hydrolysis of a caseinate through a protease. A preferred raw material is a fresh milk whey pre-concentrated to make it, preferably at 10-23 wt.%, And from fully sourced or fully deionized, ie, free from cations and free of anions. Another preferred material is a lactose-free and cation-free sweet whey protein concentrate. These starting materials can be provided in liquid form or powder form, and in the only case, they are dispersed in water, preferably demineralized with a view to their subsequent treatment. These starting materials can be derived from the milk of ruminants, such as cows, goats, sheep or buffaloes. According to a first embodiment of the process, the liquid starting material is contacted with a weakly anionic resin in a reactor, with moderate agitation, at a temperature of "_ = 50 ° C, preferably between 0 and 15 ° C. The agitation should be sufficient for the flow of resin bed. It can be produced, for example, by means of a stirrer or, preferably, by the introduction of a fluid stream, for example of air or nitrogen under pressure through the bottom of the reactor. ~~~ It is possible to use any anion exchange resin whose matrix is hydrophobic and in which the exchange groups are weakly basic in the form of gel, macroporous or macrotransparently crosslinked, preferably polystyrene or polyacrylic, particularly on the basis of a 1 poie / en vinylbenzene copolymer and preferably cross-linked more often because of osmotic shock resistance considerations The active groups are generally primary to tertiary amines. alkaline form (hereinafter referred to as OH form ") and therefore its active sites should preferably be" greatly regenerated in this form. before this contact process, the active sites of the resin are exchanged against the GMP molecules, producing a gradual increase in the pH of the treated liquid, up to a final stabilized value, for example, from 4.5 to 5.5, depending on the material of used game. The duration of the operation and the respective amounts of the resin and the treated liquid is selected as a function of the composition of the starting material and the desired amount of GMP. This operation lasts from 1 to 10 hours, for example about 4 hours. The respective proportions of resin and liquid that will be treated can vary widely and are, in volume, from 1: 1 to 1:30 and preferably from 1: 1 to 1:10., depending on the desired degree of separation of the GMP. According to another embodiment, the liquid can be percolated through a column filled with resin, the treated liquid collected therefrom and the GMP adsorbed on the resin recovered by elution. To do this, the procedure can be carried out continuously or smi nally, for example, by extracting the saturated resin from the column counter-clockwise and replacing it with freshly generated resin. The preceding embodiments, in a reactor and in a column, can be combined, for example, using a mixed device whose upper part is a reactor provided with means for stirring or for producing a fluidized bed containing the resin, separated by a grid or a filter of a lower part consisting of a column where, at the end of the treatment, the resin can be recovered, for example, by decanting, and the treated liquid re-extracted. The liquid thus treated can be concentrated, for example, by evaporation and then dried, for example, by spray drying in a drying tower. In this way, the powder obtained advantageously serves as a protein starting material in the preparation of baby products, and is important due to its desired amino acid profile, its aminogram showing a reduction in threonine and an enrichment in aromatic amino acids such as tryptophan. To separate the GMP from it, the resin is first treated by washing, for example, with demineralized water and then, when appropriate, with a diluted saline solution or a dilute acid solution and rinsed with demineralized water. The actual desorption of the GMP is carried out with an aqueous solution of acid, base or salt, preferably with a solution to basic solution, for example, NaOH, KOH or Ca (OH) 2, advantageously with a concentration of <8% by weight, preferably from 0.5 to 3%, followed by washing with demineralized water. In this way, the resin is regenerated at the same time. The eluted product and the washings are then combined and then demineralized, for example, by "ultrafiltration on anoxylation on a membrane with an average cut region of about 3000 daltons and the retention product is dry, for example, through freezing. "The GMP thus obtained is substantially free of grase and lactose and has a low whey protein content. Preferably it contains by weight: < 1% fat, < 0.2% lactose, and < 3% of true whey proteins. GMP can be used in its known applications, for example, for its biological properties in oral, parenteral or subcutaneous pharmaceutical compositions such as antimicrobial agents, antibacterial or antibacterial agents or preferably as an anti-plaque agent. and against caries in compositions for dental hygiene, or alternatively in foods, for example, confectionery products for their properties against plaque and against caries, for their functional properties as "emulsifying, gelling or foaming agents or for their dietetic properties, for example in infant compositions, as they do not contain phenylalanine. An important advantage of the process: according to the invention is that there is no reduction in the operation of the resin or scale of the same, even after 150 cycles of treatment. The following examples illustrate the invention, as well as Figure 1 of the drawing, which shows schematically and not in the form of limitation, a preferred device for carrying out the invention. In the cases, the parts and percentages are by weight unless otherwise stated.

Example 1 For the treatment, a reactor 1 was used consisting, in its upper part, of a main tank 2 communicating in its lower part with a "corff rt imen to 3" with a diameter smaller than that of the tank 2. Tank 2 is provided with a rinsing liquid inlet channel 4, a gas inlet under pressure 5, a safety valve 6 that allows the gas pressure in reactor 1 to be regulated. from its base, the tank 2 is provided with a strainer 7 and a channel for withdrawing liquid 8. At the level of the compartment 3, the reactor is provided with a pH meter 9, a gas inlet 10 and communicates through a three-way valve 11 with an inlet channel 12 for the liquid to be treated and a discharge channel 13 for the treated liquid At the base of the compartment 3, a grid or a perforated plate 14 is provided whose role is to collect the pearls of resin 15. Under grid 14, a Extraction channel 16 carries the liquid through the pump 17 to a regulating tank 18 provided with a level control device 19 and thence to the -channel 20 through "the pump 21. The channel 20 is already connected either to channel 12 or discharge overflow 22. A whey sweet whey protein concentrate, conventionally treated through electro-analysis and free of cations in a strong cationic resin, was dispersed in deionized water so that the solution had a dry matter content of 6.5%. The concentrate has the following composition: Q_ O Proteins (GMP 76 included) Lactose 4.8 Ash 2.5 Limes 8 Water rest for 100 127 kg of the dispersion were received, with an initial pH of 4.25, at the temperature of 12 ° C, through channel 12 to reactor 1 through which base air was introduced by bubbling at compartment level 3, through inlet 10 through a non-return valve 23, in order to create a fluidized bed of beads of resin 15, comprising 23 kg of weak anionic resin with a hydrophobic matrix based on polystyrene (IMAC HP 661®, Rohm &Haas, regenerated in the form of OH "). The resin beads 15 were stirred for 4 hours in contact with the dispersion due to the turbulence created by the fluidization.The pH was constantly controlled through the pH meter 9. The stabilization of the pH 5.08 indicates the end of the reaction.The air supply in 10 was then cut and the air was introduced through the upper part of the reactor - at 5, above the liquid level 24, which has the effect of pushing the liquid and sedimenting the resin beads at the bottom "3 of the reactor 2, where they are retained by the grid 14. The treated liquid is extracted by gravity through the channel 8 and through the channel "16 by means of the pump 17 to the regulating tank 18 and is discharged via the channel 20 through the -pump 21- -and further to the exit through-- channels 12 and 13. After concentration of the liquid at a dry matter content of 28% by evaporation, the concentration was spray-dried in a drying tower (these operations are not shown). Analysis of the concentrate by high performance liquid chromatography (HPLC) shows that the reaction removed 91% of the starting GMP. In addition, the powder contains 95% of the starting whey proteins. To recover the GMP, the reactor and the resin were washed with deionized water starting with channel 25 and valve 26, then channel 4 through the reactor to the outlet med. "To channels 12 and 13. The GMP was eluyó through "of the same circuit twice with 40 liters of the aqueous solution to 2% of NaOH distributed by channel 27 and valve 28 and the rinse was performed with 30 liters of deionized water After having combined the eluted product and the rinsing volumes, all concentrated to a "25 liter volume through ultrafiltration or nanofiltration with a membrane" having a nominal cut of 3000 daltons, and then the retention product was dried by freezing (these operations are not shown) and _ 750 g of the GMP were obtained, corresponding a yield of 82% in relation to starting GMP. Periodically, the resin was subjected to acid regeneration after alkaline regeneration once the equivalent of 10 resin bed volumes was treated. To do this, after elution of GMP with the alkaline solution as described above, a concentrated aqueous solution of HCl was supplied through the channel 29 and the valve 30 respectively for the resin. It was converted to the OH form by passing a concentrated aqueous solution of NaOH from the channels 27, respectively 25 for the water, then 4 and then leaving the reactor 1 through channel 16 and collecting by the pump 17 to the regulating tank 18, and then to "through" the pump 21 and was discharged by the channel 20 and "the overflow 22 to the effluent treatment." After this operation, the resin is ready for another cycle of treatment.

Example 2"Sweet bovine whey, which was previously concentrated to a dry matter content of 17%, and then demineralized through electrodialysis, free of cations in a strong cationic resin column, was released from - anions in a column of weak organic resin and dried by spray in a drying tower, with the following composition: O. or Proteins (GMP 11.7 included) Lactose 81.7 Ash 1 Lipids 1 Water rest for 100 This demineralized whey powder was solubilized in demineralized water. After removal of the cation, the solution had an initial pH of 3.8. In the "preceding plant, 392 kg of this solution was treated at the temperature of 8 ° C, while being stirred in the reactor in the presence of 23 kg of weak anionic resin of the hydrophobic matrix based on polystyrene (IMAC HP 661®, Rohm &Haas, regenerated in the OH form ") for 4 hours. Stabilizing the pH to 4.89 indicates the end of the reaction. The liquid was then removed and the resin recovered as before. After concentration of the liquid at a dry matter content of 45% by evaporation, the concentrate was spray-dried in a drying tower. dry matter of 18%, was released from the cation through treatment in a column of strong cationic resin, whose initial pH was 1.09. In the previous plant, 70 kg of this whey was treated at the temperature of 25 ° C, while stirring in the reactor in the presence of 14 kg of -the weak anionic resin of the hydrophobic matrix based on polystyrene (IRA 96 ®, Rohm &Haas, regenerated in the form of OH ") for 4 hours.L stirring was provided by- the creation of a fluidized bed of resin beads, with nitrogen bubbling.The pH stabilization at 4.79 indicates the end The liquid was then separated from the resin as above, and after concentration of the liquid at a dry matter content of 45% by evaporation, the concentrate was spray-dried in a drying tower. of the powder through HPLC shows that the reaction removed 85% of the starting GMP, however, the powder contained 9.2% of the whey proteins, corresponding to a yield of 90% of the proteins of sue r "ode milk.

The analysis of the aminogram of the concentrate shows a profile, which was characterized by a reduction of 28% in threonine, an increase of 18% in arginine and an increase of 20% in tryptophan. To recover the GMP, the resin was washed successively with deionized water, with 50 liters of an aqueous solution at 0.05% NaCl and twice with 50 liters of desi or ized water, and then the GMP was eluted twice with 25 liter of an aqueous solution at 0.6% KOH and the rinse was carried out with 10 liters of deionized water After combining the eluted product and the rinsing volumes, everything was concentrated to a volume of 25 liters through ultrafiltration with a membrane having a nominal cut-out of 3000 daltons, and then the retention product was freeze-dried, and 176 g of the GMP were obtained, running at an 80% yield with reference to the starting GMP.

EXAMPLE 4 An ultrafiltration permeate powder was used as the starting material of sweet milk, liberated from most of its salts, the composition of which is as follows: or. o Proteins (GMP 2.75 included) Lactose > 90 Ash 1.5 Water remaining for 100 The preceding powder was dissolved in demineralized water so that the solution had a dry matter content of 19.35%. This solution was released from cations through the passage on a cationic resin column -Tuerte (IR 120® Rohm &Haas), which led to a solution containing 18.73% dry matter whose pH was 2.77. 565 g of this solution and 56.5 g of the weak anionic resin of hydrophobic matrix based on polystyrene (IMAC HP 661®, Rohm &; Haas, regenerated in the form of OH ") for 3 hours at 10 ° C until the pH was stabilized to a final value of 4.53, The permeate was then" treated "after being separated from the resin beads by means of a liquid. The analysis of its aminogram shows a profile characterized by an increase of 20% in threonine and a 50% increase in tryptophan- To recover the GMP, the resin was washed with 1 liter of deionized water, and then the GMP was eluted with 50 ml of an aqueous solution at 0.6% NaOH, and then performed in "rinse with 20 ml of deionized water. After combining the eluate and the rinsing volumes, everything was concentrated through ultrafiltration with a membrane-having a nominal cut-off of 3000 daltons, and then the retention product was freeze-dried and 870 mg of GMP was obtained.

"---" Example 5 3.5 liters of sweet whey, preconcentrated to a content of matter is 20%, released from cations in a strong cationic resin column and at a pH of 1.09, percolated through a column containing 450 ml of a weak anionic resin of a polystyrene-based hydrophobic matrix (IMAC HP 661®, Rohm &Haas) at a rate of 2 bed volumes / hour The equivalent of 4 bed volumes is rec erar or "ñ", with "s replacing 4 equal fractions with a pH ranging from 6 to 3, and where the amount of GMP removed scales of 50 to 9% (evaluated through HPLC). After combining the 4 fractions, a solution with a pH of 4.5 was obtained in which 25% of the GMP was removed (compared to the whey material of p a rt ida). To recover the GMP, the procedure was carried out as in Example 1 and equivalent results were obtained with respect to the purity of the GMP.

Claims (11)

1. A process for the 'ion exchange treatment of a liquid lactic raw material containing GMP, with the aim of recovering, on the one hand, a product that can be used as a source of protein and, on the other hand, the GMP in pure form, characterized in that it comprises the following steps: i) of s ca ti oni zation of the liquid starting material, so that the pH has a value of 1 to 4.5, ii) bringing the liquid in contact with a anionic resin of hydrophobic matrix, predominantly in alkaline form up to a stabilized pH, iii) separation of the resin and the liquid product that is recovered, and iv) deactivation of the GMP of the resin ".

2. The process according to claim 1, characterized in that the starting material is a pre-concentrated sweet whey for making cheese, preferably at 10-23% by weight and cation-free or completely de-ionized .

3. The process according to claim 1, characterized in that the starting material is a sweet whey protein concentrate free of.-Lactose free and cation-free.

4. The process according to claim 1, characterized in that the starting material is the hydrolysis product, through "a protease, a native casein obtained by the acid precipitation of milk" skimmed with an inorganic acid or through acidification biological, where it is appropriate with the addition of calcium ions, the product of hydrolysis, through a protease, of a micellar casein, obtained for example, by my cr ofi 11 ration of skim milk, or alternatively the product of hydrolysis of a caseinate through a protease.

5. The process according to claim 1, characterized in that the starting material is a permeate of the ultrafiltration of the sweet whey.

6. The process according to claim 1, characterized in that the liquid starting material is contacted with a weak anionic resin predominantly in alkaline form in a moderately stirred reactor at a temperature of < 50 ° C, preferably between "0 and 15 ° C for one or several hours, which produces a gradual increase in the pH of the treated liquid, until stabilization is obtained and then the liquid is separated from the resin by ultrafiltration or centrifugation - - - Z

7. The process according to claim 6, characterized in that the weakly basic anion exchange resin in the form of macroporous gel or interlaced ma crotr ans rsa lmen te, whose matrix "Being hydrophobic, it is used.

8. The process according to claim 6, characterized in that the liquid thus treated, in particular by evaporation, is concentrated and then dried, in particular spray-dried in a drying tower. 9. The process according to claim 1, characterized in that "the GMP is separated from it in purified form, the resin is first treated by washing, and then the GMP is desorbed with an aqueous acidic, basic or saline solution, in particular NaOH, KOH or Ca (OH) 2, of concentration <8%, and rinsed with demineralized water, the eluted product and the rinses are then combined and demineralized, in particular through ultrafiltration or nano fi ltration in a membrane with a "middle region of approximately 3000 daltons, and the retention product is dried, in particular freeze-dried. 10. The use of purified GMP, which can be obtained through the process according to the indication 9, to prepare an antimicrobial, pharmaceutical, antimicrobial, antimicrobial composition. concentrated and then dried, in particular spray-dried in a drying tower.

9. The process according to claim 1, characterized in that the GMP is separated from it in purified form, the resin first treated by washing, and then the GMP is desorbed with an aqueous acidic, basic or saline solution, in particular NaOH, KOH or Ca (OH) 2, concentration <8%, and rinsed with demineralised water, the eluted product and the rinses are then combined and demineralized, in particular through ultrafiltration or nano fi ration in a membrane with a mean region of approximately -3000 daltons. , and the retention product is dried, in particular freeze-dried.

10. The use of purified GMP, which can be obtained through the process according to claim 9, for preparing an antimicrobial, antimicrobial, antibacterial, pharmaceutical composition.

11. The use of purified GMP which can be obtained through the process according to claim 9 for preparing a dental hygiene composition against plaque and against cavities.