BE1026978A1 - PROCESS FOR THE PRODUCTION OF OVOMUCOIDS AND OVOINHIBITORS FROM A LOT OF COTURNIX COTURNIX JAPONICA QUAIL EGGS - Google Patents

Abstract

Process for the production of ovomucoids and ovoinhibitors from a batch of Cotunix cotunix japonica quail eggs from a homogenate of concentrated whites and yolks, the drying of which is optimized.

Description

The present invention relates to a process for the production of ovomucoids and ovoinhibitors from a batch of eggs COTURNIX COTURNIX JAPONICA The present invention relates to a process for the production of ovomucoids and ovoinhibitors from a batch of eggs Quail Coturnix coturnix japonica.

quail egg has long been recognized for its therapeutic virtues. Several clinical studies have been devoted to it and, more specifically, concerning its antiallergic properties to treat asthma, allergic conjunctivitis, eczema and other pathologies.

Coturnix coturnix japonica quails are the domesticated form of wild quail (Coturnix coturnix) whose eggs are particularly rich in albumen. They should not be confused with the species of Asian origin, known as the Japanese quail (Coturnix Japonica). Egg white contains active substances known for their antiallergic properties such as ovomucoids and ovoinhibitors. The antiallergic activity depends on the fraction of ovomucoides and ovoinhibitors present in the egg white. Egg white also has anti-inflammatory properties. Ovomucoid is a protease inhibitor enzyme found in the whites of bird eggs. More specifically, ovomucoid is an enzyme that inhibits human serine proteases. This enzyme is a polypeptide which comprises three very similar functional domains each capable of binding a protease. For example, the quail egg comprises a polypeptide essen ASS which can bind trypsin and also activate it. This is called anti-tryptic activity. Ovainhibitor is a glycoprotein which is structurally different from that of ovomucoide. Vovoinhibitor is also part of ‘protease inhibitors and more specifically, serine proteases. The ovoinhibitor has a potent activity against Velastase which is involved in numerous pathologies such as pulmonary emphysema or psoriasis.

Proteases are enzymes endowed with proteolytic pro-inflammatory properties contained in the intracytoplasmic granulations of immune cells. Certain proteases, and more particularly serine proteases, are involved, in humans, in physiological processes such as [a coagulation, complement activation and immunity. Serine proteases are involved in the activation cascade of certain immune cells such as hasophils and mast cells.

On the other hand, for certain pathologies, the proteases released by the pro-inflammatory cells can lead to a worsening of the symptoms, for example for asthma. fl is therefore important to achieve a protease / anti-protease balance in order to alleviate certain symptoms. An intake of protease inhibitors, for example, via an ovomucoid or an ovoinhibitor can therefore alleviate certain symptoms of pathologies.

In particular, the invention relates to a process for producing ovomucoids and ovainhibitors from a batch of Coturnix coturnix japonica quail eggs comprising the steps of: {a) washing and disinfection of said batch of eggs Quail Coturnix coturnix japonica;

b} opening of the shells of each egg of said batch of Coturnix coturnix japonica quail eggs with recovery of the components of the egg without said shell: c} separation of the components of Egg with recovery of a fraction of egg whites and a fraction of egg yolks: This process is known from document EP 315 552, according to law, a stock solution of Coturnix coturnix japonica quail egg yolks and whites is formed, diluted in water to impregnate granules which are then , dried.

The documents FR 2 836 379, FR 2 813 531, CH 695 925, FR 2 811 19 898 describe the same process except that additives are added, in addition, to the granules, More precisely, each additive is added by additional impregnation of granules by said additive.

In addition, these granules are final formulations intended for the consumer or the patient. The present invention more particularly aims at the production of a powder concentrated in active material in order to develop different final formulas according to the markets and the specific requests of formulators.

The process according to these documents, if it is aimed at a solid formulation obtained, it does not make it possible to obtain a powder as a final or intermediate product, nor does it make it possible to obtain a concentrated product when the stock solution is very diluted to comply with the conditions of homeopathy.

There is therefore a general need for a method which allows the production of ovomucoids and ovoinhibitors from Coturnix coturnix japonica quail eggs in high yield, while retaining the therapeutic properties of ovomucoids and ovoinhibitors.

The present invention intends to solve the problem by a method for producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs characterized in that, the method further comprises the steps of: in contact with a predetermined fraction of egg fauna, in which the egg yolk is the major component, with a predetermined fraction of egg white, in which the egg white is the major component in a proportion of '' at least two whites for one yolk, with formation of a mixture of whites and yolks: e} homogenization of said mixture of whites and yolks of said batch of Coturnix coturnix japonica quail eggs with obtaining a homogenate of whites and of concentrated yolks: f) Feeding a mixer with microcrystalline cellulose and subsequent feeding of the homogenate of concentrated whites and yolks with obtaining a fluffy powder of said microcrystalline cellulose and of said homogenate whites and yolks; g) Mixing for a predefined mixing time T of said microcrystalline cellulose with the homogenate of concentrated whites and yolks.

h} drying of said fluffy powder in an enclosure at a drying temperature of between 80 ° C and 85 ° C, preferably between 81 ° C and 84 ° C, more particularly at 82 ° C, and for a period of between 6 hours and 8 hours, preferably for 7 hours, obtaining a crude dried powder of ovomucoids and ovoinhibitors in a homogeneous mixture.

By the expression two whites for one yolk is meant by way of example the addition to all of the egg whites of half by weight of the previously separated egg yolks. Alternatively, by way of example is also meant the addition to n whole eggs (white + yolk) of n egg whites.

In the process according to the present invention, it has been observed, against all expectations, that drying the mixture of microcrystalline cellulose with the homogenate of concentrated whites and yolks makes it possible to obtain a non-sticky, homogeneous and dried powder of optimally with a maximum of 5% humidity, without degrading the ovomucoids and ovoinhibitors, without having to require a subsequent sterilization / pasteurization step, and without having to resort to dilutions in order to overcome the high viscosity of the homogenate.

According to the documents cited above, the drying is carried out post-impregnation of the granules and imperatively requires serial dilutions in order to allow the impregnation of the granules with the relatively viscous homogenate. In addition, the drying is carried out at around 40 ° C so as not to denature the ovomucoids and ovoinhibitors, but also because the homogenate is diluted with an aqueous solution allowing it to be dried at around 40 ° C.

Although it is otherwise known to dry ovomuroids and ovoinhibitors by lyophilization, when a large amount of quail egg homogenate has to be dried, it is not possible to carry out such lyophilization effectively. . Lyophilization of the homogenate of quail eggs unfortunately forms a sticky or even viscous powder which cannot be used directly. In addition, the lyophilization cycles are long and can last from 20 hours to 72 hours. The product must then be further sterilized or pasteurized for further use in the pharmaceutical, nutraceutical or food supplement fields.

According to the present invention, it is possible to obtain a mixture of non-coflating but homogeneous powder in which the active substances retain their activity without having to resort to slow lyophilization, which slow lyophilization further requires a pasteurization / sterilization step and requires extremely expensive equipment. Drying, according to the present invention, the temperature of which is between 80 ° C and 85 ° C, and for a period of between 6 hours and 8 hours makes it possible to eliminate the bactericological risk while making it possible to maintain optimal activity of the bacteria. ovomucoids and ovoinhibitors without investment in expensive equipment, thus ensuring optimized process efficiency.

In fact, unlike the drying at 40 ° C, which may be sufficient post-impregnation of a small amount of egg homogenate diluted on a granule, the drying according to the invention, the duration of which is only 6 hours at 8 hours and at a temperature between 80 ° C and 85 ° C, it was possible to dry a large amount of viscous egg homogenate in a relatively short time. In addition, since the temperature remains below 85 ° C, the ovomucoids and ovoinhibitors are not degraded while being pasteurized.

1 has therefore been surprisingly demonstrated that the narrow window of temperature and time makes it possible to obtain a non-sticky powder mixture which can be used directly. Indeed, said powder mixture can be packaged using a tablet machine or a capsule filler or, alternatively, used as an additive. This, while preserving the active substances and allowing long-term storage because the powder is pasteurized.

In another preferred embodiment according to the present invention, the process for producing ovomucoids and ovoinhibitors is characterized in that said homogenate of whites and yolks consists of whites and yolks of quail eggs Coturnix coturnix japonica . Indeed, this makes it possible to keep the initial concentration of each constituent and an optimal activity of the ovomucoids and ovoinhibitors.

; BE2019 / 5039 In a variant according to the invention, the method for producing ovomucoids and ovoinhibitors is characterized in that a dosage and adjustment of the concentration of ovomucoids and of ovoinhibitors are carried out in order to reach a predefined concentration with obtaining of a standardized extract. The dosage and adjustment make it possible to obtain a product whose concentration of ovomucoids and ovoinhibitors is known and which meets specific requirements.

More particularly, the method of producing ovomucoids and ovoinhibitors according to the invention is characterized in that the adjustment of the concentration of ovomucoids and ovoinhibitors is carried out by adding microcrystalline cellulose.

Advantageously, the method for producing ovomucoids and ovoinhibitors according to the invention is characterized in that the disinfection of the quail eggs is carried out by a predetermined number of sprays of a disinfectant solution, the duration of each vaporization varying and being between 1 minute and 15 minutes, preferably between 1.5 minutes and 12 minutes, more preferably 2 minutes and 10 minutes. Each spraying of the eggs placed on a shelf is carried out continuously to allow homogeneous and complete coverage of the surface of the egg by the disinfectant solution.

In a variant according to the present invention, the process for producing ovomucoids and ovoinhibitors is characterized in that the opening of the shells is carried out in the upper part of the shell and by a precise breaking tool, for example scissors. to cut the quail eggs. Indeed, the use of a precise breaking tool makes it possible to avoid a perforation of the yolks, which is very important given that the homogenate of egg whites and yolks is produced from a ratio of at least 2 whites for a yolk, after separating the whites and yolks while properly breaking the membrane of the quail egg.

ë BE2019 / 5039 Advantageously, the method for producing ovomucoids and ovoinhibitors according to the invention is characterized in that the homogenization of the mixture of yolks and whites is carried out using a mixer-beater, the duration of said homogenization being between 1 minute and 5 minutes, preferably 2 minutes; with a rest time of between 1 minute and 15 minutes, preferably between 8 minutes and 12 minutes, more preferably 10 minutes.

More advantageously, the method of producing ovomucoids and ovoinhibitors according to the invention is characterized in that the ratio of the mass of microcrystalline cellulose to the mass of said homogenate of egg whites and yolks is between 1 and 2 , preferably between 13 and 1.7, more particularly around 15. This ratio will make it possible to obtain a non-viscous flaky powder.

Preferably, the process for producing ovomucoids and ovoinhibitors according to the invention is characterized in that said predefined mixing time T is between 1 minute and 15 minutes, preferably between 3 minutes and 7 minutes, more preferably, 5 minutes.

In fact, this mixing time makes it possible to obtain a non-viscous fluffy powder.

More preferably, the process for the production of ovomucoids and ovoinhibitors according to the invention is characterized in that said subsequent feeding of concentrated white and yolk homogenate is effected by several additions staggered over time, between which an intermediate mixing takes place during a period of time between 30 seconds and 3 minutes, preferably between 1 minute and 2 minutes, more preferably about 1 minute.

Indeed, this mixing process makes it possible to obtain a non-viscous fluffy powder.

In another variant according to the invention, the method for producing ovomucoids and ovoinhibitors is characterized in that an additional step of drying said crude dried powder in said chamber at a drying temperature of between 80 ° C and 85 ° C. ° C, preferably between S1 ° C and 84 ° C, more particularly at 82 ° C, and for a period of between 30 minutes and 2 hours, preferably for 1 hour. In the case where the water content of the crude dried powder is greater than or equal to 5%, this additional drying step makes it possible to obtain a crude dried powder whose water content is strictly less than 5%.

In another preferred embodiment of the invention, the process for producing avomucoids and ovoinhibitors is characterized in that a step of sieving said crude dried powder of ovomucoids and ovoinhibitors is carried out by means of a calibrated sieve with a mesh of between 100um and 500um, preferably between 200um and 300um, more preferably 250um. This results in a calibrated powder of ovomucoids and ovoinhibitors that can easily be used alone or with additives.

Other embodiments of the process for producing ovomucoids and ovoinhibitors are indicated in the appended claims.

Other characteristics, details and advantages of the invention will emerge from the description given below, without limitation and with reference to the appended examples.

Example 1: protocol for the production of a powder of ovomucoids and 14ke ovoinhibitors from a batch of quail eggs Coturnix coturnix japonica 1800 quail eggs were placed on 11 slatted shelves for eggs of quail in a laundromat. Each shelf has a storage capacity of 170 quail eggs that do not overlap. 170 eggs were placed on each of the first 10 shelves. The remaining eggs have been placed on the 11th shelf. This 11) "ciayette is a backup shelf if quail eggs from

First 10 should be discarded after washing or disinfecting or opening the shells.

Each shelf of quail eggs was washed with potable veal with a temperature above 35 ° C for at least 1 minute. > Then, each shelf of quail eggs was sprayed for the first time continuously with a disinfection solution for at least 2 minutes. Each shelf of quail eggs was sprayed continuously for a second time with disinfection solution for a minimum of 10 minutes. Broken or cracked eggs are discarded. The intact eggs were rinsed in cold potable water for a minimum of one minute and drained for a minimum of 5 minutes.

11 shelves containing all the eggs intact were then placed on a means of transport such as a trolley made up of different levels, one shelf being arranged on one level. This means of transport is protected by surrounding the sides as well as the upper part by a means of protection such as protective plastic film. The means of transport containing the shelves were then taken to the ovomucoid and ovoinhibitor production room.

In the production room, the eggs were separated into 2 groups, the first group consisting of 850 eggs as well as the second group. A first bucket which serves as a container for the egg yolks of the first group and a second bucket which serves as a container for the whole eggs of the second group were tared. The shells of the first group of eggs were opened with the help of a precise breaking tool. The separation of the whites and the yolks was carried out by a device for the separation by gravity of the whites from the yolks. The yolks were received in the first tared bucket and the whites in another container, such as, for example, a stainless steel tank. The mass of the yolks was weighed.

The shells of the second set of eggs were opened using a precise breaking tool. The whole eggs were directly recovered in the BE2019 / 5039 second tared bucket and their mass was weighed. Said whole eggs are then added to the whites which are in another container, such as, for example, a stainless steel tank. The mixture of whole eggs and egg whites was homogenized using a mixer-beater for about 2 minutes. The homogenate was then allowed to stand for about 10 minutes. The mass of the homogenate was weighed. The homogenate of egg whites and yolks was then separated into two equal amounts in two pre-tared buckets. The mass of microcrystalline cellulose is calculated as a function of [a total mass of the homogenate, said total mass of the homogenate being the addition of the 2 masses of homogenates present in the 2 buckets. The ratio of the mass of microcrystalline cellulose to the said total mass of the homogenate of egg whites and yolks is 1.5. The calculated mass of microcrystalline cellulose was poured into a mixer. Said homogenate of egg whites and yolks was then poured in in different stages. Between each pour of said homogenate of egg whites and yolks into the mixer, about 1 minute was mixed. When the last step is carried out, that is to say the last payment of said homogenate of egg whites and yolks, was mixed for about 5 minutes to obtain a flaky powder. Said flaky powder was then placed on a total of 32 trays, each tray containing 750g of said fluffy powder. These trays were protected with a protective film and transported to the drying room.

We turned on the drying cabinet which was set at 82 ° C for a drying time of 7 hours. The protective film has been removed from the trays. When the temperature reached 82 ° C, the trays were placed in the drying chamber. The 7h drying time was triggered when the temperature in the drying chamber rose to 82 ° C. When the drying time of 7 hours was reached, the drying chamber was turned off and the trays unloaded. A crude dried powder was obtained from which a sample was taken to measure the water content of the sample. If the water content of the sample is greater than or equal to 5%, the crude dried powder is again heated to 82 ° C for one hour, The crude dried powder was then poured into a double bag which is placed in a was previously tared. The double pre-tared bag was then closed as well as the barrel. The mass of the drum containing said crude dried powder enclosed in the double bag was then weighed.

Finally, said crude dried powder was transported to the calibration room. Said crude dried powder was sized using a sizer and a 250um mesh sieve. The calibrated powder is received in tared plastic bags and is then weighed. Samples were taken for microbiological analysis of the calibrated powder.

A plastic bag was then tared in which all of the remaining calibrated mass was packed. The mass of the conditioned powder corresponds to the mass of the calibrated powder minus the mass of the samples. The bag is then closed and placed in another bag present in a drum of more than 60 L. The drum is then closed with a lid and labeled.

Example 2 Concentrations of Ovomucoids per 100g as a Function of Drying Time Ovomucoids and ovoinhibitors were produced from 9 batches of quail eggs according to the process according to the present invention by varying the drying time, the number of quail eggs used being equal for each batch,

| Number | Time of | Ovomucoids | lot | drying wed / 190g I 4h 5) 13 14h 29.8 | 4 0 14h | 24.6 | LM | 160 | 6 I 9 | 249 | The above table shows that when the drying time is 14 h, the concentration of ovomucoids in the raw dried powder drops sharply.

This decrease is due, among other things, to a degradation of the ovomucoids.

However, when the drying time is 7 hours, according to the present invention, the concentration of ovomucoids remains high.

Regarding batches 5 to 9, the variability of the concentration of ovomucoids with respect to the drying time also depends on the mass of egg whites in each batch.

In fact, ovomucoids are found in the white and the richer the white is in proteins (ovomucoids, ovoinhibitors), the higher their concentration.

This example 2 therefore shows the dependence of the concentration of ovomucoids on the drying time of the different batches of Coturnix coturnix japonica quail eggs. It is understood that the present invention is in no way limited to the embodiments described above and that many modifications can be made thereto without departing from the scope of the appended claims.

Claims (1)

4. A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs comprising the steps of: a) Washing and disinfection of said batch of Coturnix coturnix japonica quail eggs; b} opening of the shells of each egg of said batch of Coturnix coturnix japonica quail eggs with recovery of the components of the egg without said shell: c} separation of the components of the egg with recovery of a fraction of egg whites and a fraction of egg yolks; characterized in that, the method further comprises the steps of: d} contacting a predetermined fraction of egg yolk, in which the egg yolk is the major component, with a predetermined fraction of egg yolk. eggs, in which the egg white is the major component in a proportion of at least two whites to one yolk, with formation of a mixture of whites and yolks: 8) homogenization of said mixture of whites and yolks of said batch of Coturnix coturnix japonica quail eggs with obtaining a homogenate of concentrated whites and yolks: f) Feeding a mixer with microcrystalline cellulose and subsequent feeding of the homogenate of whites and yolks concentrated with obtaining a fluffy powder of said microcrystalline cellulose and of said homogenate of whites and yolks: g) Mixing for a predefined mixing time T of said microcrystalline cellulose with the concentrated homogenate of whites and yolks. h} drying of said fluffy powder in an enclosure at a drying temperature of between SOC and 85 ° C, preferably between 81 ° C and 84 ° C, more particularly at 82 ° C, and for a period of between 6 hours and 8 hours, preferably for 7 hours with obtaining a crude dried powder of ovomucoids and ovoinhibitors in a homogeneous mixture. 2, A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to claim 1, wherein said homogenate of whites and yolks consists of egg whites and yolks. Quail Coturnix coturnix japonica. 3. A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to claim 1 or 2, in which dosage and adjustment of the concentration of ovomucoids and ovoinhibitors are carried out. to reach a predefined concentration with obtaining a standardized extract. 4, A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to any one of the preceding claims, in which the adjustment of the concentration of ovomucoids and ovoinhibitors is carried out by the addition of microcrystalline cellulose. 5. A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica eggs according to claim 1, wherein the disinfection of the quail eggs is carried out by a predetermined number of sprays of a disinfectant solution, the duration of each vaporization being between 1 minute and 15 minutes, preferably between 1.5 minutes and 12 minutes, more preferably 2 minutes and 10 minutes. 6. A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to claim 1, wherein opening of the shells is carried out in the upper part of the shell and by a precise tool. breaking, for example scissors to cut the quail eggs. 7, A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to claims 1, 5 or 5, wherein the homogenization of the mixture of yolks and whites is carried out at using a beater mixer, the duration of said homogenization being between 1 minute and 5 minutes, preferably 2 minutes; with a rest time of between 1 minute and 15 minutes, preferably between 5 minutes and 12 minutes, more preferably 10 minutes. A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to claim 1, wherein the ratio of the mass of microcrystalline cellulose to the mass of said homogenate of whites and whites. egg yolks is between 1 and 2, preferably between 1.3 and 1.7, more particularly around 1.5, 9, Process for producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to claim 1, wherein said predefined mixing time T is between 1 minute and 15 minutes, preferably between 3 minutes and 7 minutes, more preferably 5 minutes. 10. A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to claim 1, wherein said subsequent feeding of the concentrated white and yolk homogenate is carried out by several. additions staggered over time, between which an intermediate mixing takes place for a period of time between 30 seconds and 3 minutes, preferably between 1 minute and 2 minutes, more preferably about 1 minute. 11. A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to any one of the preceding claims, wherein an additional etane for drying said crude dried powder in said chamber. at a drying temperature of between 80 ° C and 85 ° C, preferably between 81 ° C and 84 ° C, more particularly at 82 ° C, and for a period of between 30 minutes and 2 hours, preferably for 1 hour . 12. A method of producing ovomucoids and ovoinhibitors from a batch of Coturnix coturnix japonica quail eggs according to any one of the preceding claims, wherein a screening stage of said crude dried powder of ovomucoids and of ovoinhibitors is produced by means of a calibrated sieve with a mesh size of between 100 μm and 50 μm, preferably between 200 μm and 300 μm, more preferably 250 μm, with obtaining a powder of ovomucoids and of calibrated ovoinhibitors.