BG113181A - Lyophilised snail products and preparation method thereof - Google Patents

Abstract

The method and the products obtained thereof will find application in the food, cosmetic and pharmaceutical industries. The method is continuous and waste-free and includes the following stages: Stage 1 hatching of snails and/or fattening of snails, Stage 2 preparation of snail eggs, Stage 3 preparation of snail slime, Stage 4 preparation of snail haemocyanin, Stage 5 preparation of snail meat, Stage 6 preparation of snail meat protein hydrolysate, Stage 7 preparation of snail shell protein hydrolysate and Stage 8 lyophilisation process of the products obtained in Stage 2 to Stage 7. The products obtained therefrom are lyophilized caviar, lyophilized mucus, lyophilized hemocyanin, lyophilized meat, lyophilized meat protein hydrolysate and lyophilized snail shell protein hydrolysate. The advantages of the products obtained by the method are as follows: - full preservation of the biological activity of the components contained therein; - a prolonged shelf life of 2 to 5 years, without the use of preservatives; - storage and transport under standard conditions.

Description

LYOPHILIZED SNAIL PRODUCTS AND METHOD FOR THEIR OBTAINMENT

FIELD OF ENGINEERING

The method and the products obtained according to the invention will find application in the food, cosmetic and pharmaceutical industries.

PRIOR ART

A healthy product from snails and a method for its preparation is known from patent document CN1321587. The product contains a mixture of lyophilized products, including snail meat, and the remaining components are lyophilized royal jelly, galactose, dietary fiber, vitamin C. The method includes the following technological operations: removing the snail from the shell, heat treatment, grinding the meat, freezing the snail paste for subsequent lyophilization which takes place under vacuum for 24-72 hours.

Patent document CN1275 607 relates to a medicinal composition for the treatment of hyperhydrochloria, a method for its preparation and its use. The medicinal composition includes lyophilized snail meat, lyophilized snail shells, and the remaining components are: lyophilized white turnip or spinach. Its application is to inhibit stomach acid and as an anti-ulcer agent. The method for its preparation includes a lyophilization process taking place at a temperature of -40 20°C for 12-42 hours. The remaining stages are the classic ones - removing the meat, grinding, subsequent lyophilization and tableting.

A method of obtaining polysaccharides from snails is known from patent document CN10049422IC Belly bottom snail polysaccharide and its preparing method, which includes the following stages: a. Raw materials and processing: fresh snail meat is rinsed with water, followed by draining the water; b. Homogenize in a homogenizer and for 15-30 minutes to obtain a homogenate; drying at 0°C 70°C, then grind to powder, add 0.5-30 times water and mix to obtain a homogenate; Post-treatment options include: ultrasonic radiation in an ultrasonic generator and oscillation for 0-4h or/and microwave treatment 50-600w for 10 minutes; Enzymolysis: a neutral protease with an enzyme specific activity of 600-1200 u/g is added to the homogenate treated in step c, enzymolysis takes place at pH 6.5-8.5, at a temperature of 35-50°C for 1-3h, an enzyme ratio is added after destroying the enzyme pepsin with an activity of 6000-12000 u/g, continuing to hydrolyze at pH 1.0-3.5, at a temperature of 35-50°C for 1-5 hours and then adjusting to pH 6.5- 7.5, purpose of obtaining snail meat or its hydrolyzate; Water extraction: heating the hydrolysis to 70-100°C, water extraction for 0.5-5 hours, the extraction is from 1-3 times; Concentration: concentration of the aqueous extract to a solids content of 15-30%; Alcohol precipitate - Add 2 to 6 volumes of 95% by weight ethanol to the concentrated solution and, after standing for 8 to 24 hours, centrifuge at 4000 r/min for 15 minutes to collect the precipitate; To the protein is added: 0.5 to 3 times the volume of water to the precipitate, 0.1 to 0.5 times the volume, a 4:1 to 5:1 volume ratio of chloroform to amyl alcohol or n-butanol solution, vigorously shaken for 20 to 30 minutes, centrifugation and removal of the denatured protein at the interface between the aqueous layer and the solvent layer to obtain an aqueous polysaccharide solution; Alcohol precipitate: add 2 to 6 times the volume of 95% ethanol by weight to an aqueous solution of polysaccharide and, after standing for 8 to 24 hours, centrifuge at 4000 r/min for 15 minutes to collect the precipitate; Freeze drying: add 0.1 to 0.5 times water, cool to -30 to -35°C, freeze, then dry in vacuum from 50 to 80 Pa, temperature from 30 to 60°C, then dry in for 10 to 15 hours, the dry polysaccharide powder being obtained with a polysaccharide yield of 3.0-16.0% and a purity of 25-55%.

TECHNICAL ESSENCE OF THE INVENTION

The aim of the invention is to create a method for obtaining products from snails that is continuous and waste-free and that the products obtained according to it have high qualitative and quantitative indicators.

The goal is achieved with a method which, according to the invention, is continuous and waste-free and includes the following stages, stage 1 hatching of snails and/or fattening them, stage 2 obtaining caviar from snails, stage 3 obtaining slime from snails, stage 4 obtaining hemocyanin from snails, step 5 obtaining snail meat, step 6 obtaining protein hydrolyzate from snail meat, step 7 obtaining protein hydrolyzate from snail shell and step 8 lyophilization process of the products from step 2 to step 7, with step 1 being process of hatching snails and/or their fattening in controlled conditions for a period of 3-4 months, in specialized premises for the purpose, in stage 2 includes fertilization of the snails, spawning, with the snails located at a density of 250-300 pieces of m and the laying process lasts from 36 to 48 hours, stage 3 the process of obtaining mucus from snails is carried out in a vessel with equivalent amounts of water and live snails, the vessel is equipped with a stirrer of 30 to 60 revolutions per minute and elect gave birth to pulse supply of low-voltage voltage from 10 to 12 Volts, which is supplied for a time of 60-120 seconds from 2 to 3 times, after which the snails are removed from the vessel, the solution is allowed to homogenize for 10-14 hours at a temperature of 4 -6°C, filtered successively through 50, 30, 20, 10, 5, 1 micron pore size filters, after which the resulting filtrate is subjected to additional filtration for concentration, by reverse osmosis or dialysis or ultrafiltration, stage 4 the process of obtaining hemocyanin is carried out manually by making an S-shaped movement along the lower part of the leg of the snail, ending at the end with a light sting and releasing the hemocyanin, stage 5 process of obtaining meat from snails, which is carried out by thermal processing, separating the meat from the shell is carried out in water preheated to 70°C for 30-40 seconds, followed by mechanical removal of the meat and its immersion in pre-cooled water with a temperature of 20°C, stage 6 process of isolation of protein hydrolyzate from snail meat, which is carried out by enzymatic hydrolysis in a stainless steel vessel equipped with a low-speed stirrer and a heater in alkalized (pH=8) water, which has been previously heated to 60°C, the ground snail meat is added and an enzyme is added, the process takes place for 10-14 hours at a temperature of 60°C to 95°C, with continuous pH control and, if necessary, corrected with a suitable alkaline solution, followed by cooling to 20°C and acidification to pH=4.5 -5.5, step 7 the process of obtaining protein hydrolyzate from snail shell proceeds, the snail shells obtained from step 5 are washed repeatedly with chilled water, then air-dried to constant weight, followed by mechanical grinding to 5 mm size and placing them in a stainless steel vessel with a low-speed stirrer and a heater to preheated to 60°C alkalized (pH= 8) water. Enzyme is added to the mixture and the enzymatic hydrolysis process is carried out for 20-24 hours at 60°C and pH=8 of the reaction medium. The process is carried out under continuous pH control and, if necessary, corrected with an alkaline solution. At the end of the process, the temperature of the reaction mixture was raised to 95°C and maintained for 4 hours. The reaction mixture is then cooled sharply to 20°C and acidified to pH=4.5-5.5 with a suitable acid, step 8 the products obtained from steps 2, 3, 4, 5, 6 and 7 are lyophilized.

The method according to the invention offers an embodiment in which step 2 and step 3 can be repeated multiple times.

According to the invention, the additional filtration of stage 3 is carried out by reverse osmosis or dialysis or ultrafiltration.

According to the invention, the process of lyophilization of the products according to the method takes place at a temperature of minus 40-45°C for a time of 32 to 48 hours.

Products obtained by the method according to the invention are:

Freeze-dried caviar, which is a light, fine powder with a pink-reddish color, with a protein content of 25-35% and carbohydrates 7-12%, and has a content of free amino acids as follows:

Amino acid content in mg/year minimally maximum __ ΰ Histidine 0.288 0.352 Arginine 0.243 0.297 Asparagine 0.162 0.198 Glutamine 0.252 0.308 4-Hydroxyproline 0.027 0.033 Serine 0.873 1,067 Aspartic acid 0.405 0.495 Glutamic acid 0.234 0.286 Threonine* 0.216 0.264 Glycine 0.27 0.33 Gamma-aminobutyric acid - - Tyrosine 0.072 0.088

Proline 0.117 0.143 Methionine 0.018 0.022 Valine* 0.171 0.209 Leucine/Isoleucine* 0.207 0.253 tryptophan* 0.063 0.077 Phenylalanine 0.126 0.154 Ornithine 0.576 0.704 Lysine* 0.261 0.319 General content 4,581 5,599

Lyophilized mucilage, which is a fine powder with a beige-brown color, the protein fraction is 40 - 45%, and the content of free amino acids is as follows

Amino acid content in mg/year minimally maximum Histidine 0.43 0.53 Arginine 0.11 0.13 Asparagine 0.39 0.47 Glutamine 0.40 0.48 4-Hydroxyproline 0.11 0.13 Serine 0.63 0.77 Aspartic acid 0.90 1.10 Glutamic acid 1.39 1.69 threonine* 0.43 0.53 Glycine 2.40 2.94 Gamma-aminobutyric acid - - Tyrosine 0.05 0.06

Proline 0.45 0.55 Methionine 0.10 0.12 Valine** 1.64 2.00 Leucine/Isoleucine** 3.40 4.16 tryptophan** 0.19 0.23 Phenylalanine 0.82 1.00 Ornithine 1.48 1.80 Lysine 1.73 2.11 General content 17.03 20.81

# essential amino acid

Lyophilized hemocyanin, which is a fine white powder containing copper (Si) coordinated hemocyanin proteins, reaching up to 90% of the product content.

Freeze-dried snail meat is a brown powder with a protein content of 6575%, carbohydrates 14-20% and with free amino acid content as follows:

Amino acid content in mg/year minimally maximum Histidine** 0.621 0.759 Arginine 0.18 0.22 Asparagine 0.333 0.407 Glutamine 2,106 2,574 4-Hydroxyproline 0.063 0.077 Serine 1,026 1,254 Aspartic acid 0.882 1,078

Glutamic acid 2,763 3,377 threonine* 0.549 0.671 Glycine 0.864 1,056 Gamma-aminobutyric acid - - Tyrosine 0.414 0.506 Proline 0.675 0.825 Methionine 0.135 0.165 Valine** 0.756 0.924 Leucine/Isoleucine** 0.909 1,111 tryptophan** 0.162 0.198 Phenylalanine 0.603 0.737 Ornithine 7,803 9,537 Lysine** 2.16 2.64 General content 23,004 28,116

# essential amino acid

Lyophilized protein hydrolyzate from snail meat containing low molecular weight peptides 65-75%, protein 10-15%, carbohydrates 10-14% and amino acids as follows:

Amino acid content in mg/year minimally Maximum Histidine** 8.73 10.67 Arginine 2,043 2,497 Asparagine 14.76 18.04 Glutamine 30,429 37,191 4-Hydroxyproline 0.072 0.088 Serine 0.999 1,221 Aspartic acid 8,136 9,944 Glutamic acid 17,838 21,802

Threonine 0.405 0.495 G lysine 5,004 6,116 Gamma-aminobutyric acid Tyrosine 15,633 19,107 Proline 1,926 2,354 Methionine 6,084 7,436 valine" 17,055 20,845 Leucine/Isoleucine 25,713 31,427 tryptophan" 4,509 5,511 Phenylalanine 18,603 22,737 Ornithine 32,724 39,996 Lysine" 26,586 32,494 General content 237,249 289,971

# essential amino acid

Lyophilized snail shell protein hydrolyzate, which is a beige-brown powder with free amino acid and micronutrient content as follows:

Amino acid content in mg/year minimally Maximum Histidine 0.31 0.38 Arginine 0.02 0.02 Asparagine 0.18 0.22 Glutamine 2.99 3.65 4-Hydroxyproline 0.01 0.01 Serine 0.40 0.48 Aspartic acid 0.02 0.02 Glutamic acid 0.06 0.07 threonine" 0.23 0.28 Glycine 0.44 0.54

Gamma-aminobutyric acid - - Tyrosine 1.40 1.71 Proline 0.02 0.02 Methionine 0.06 0.07 Valine* 0.51 0.63 Leucine/Isoleucine 0.46 0.57 tryptophan* 0.06 0.07 Phenylalanine 0.76 0.93 Ornithine 0.10 0.13 Lysine* 1.34 1.64 General content 9.37 11.45

# essential amino acid

A trace element content in mg/year minimally Maximum Calcium 189,010 231,020 Magnesium 172,800 211,200 Chrome 0.003 0.004 Sodium 5,020 6,140 Zinc 0.101 0.123 Iron 1,066 1,303

All processes, according to the invention, in which there are manipulations with live animals are realized in strict compliance with the requirements of the veterinary authorities.

The method according to the invention allows the full utilization of all components of the snail, for which there is evidence that they possess beneficial properties in relation to human health.

The method includes the stage from hatching the snail to obtaining lyophilized products from it, without side waste affecting the environment.

The method is implemented both in specialized premises and outdoors in constructed farms related to the fattening of snails.

The method is continuous and waste-free, and from one snail in a continuous cycle, 6 final lyophilized products are obtained, which are:

• Lyophilized caviar;

• Lyophilized mucus;

• Lyophilized hemocyanin (blood);

• Lyophilized meat;

• Lyophilized protein hydrolyzate from meat;

• Lyophilized shell protein hydrolyzate.

Advantages of the products obtained by the method are:

• Complete preservation of the biological activity of the components contained in them;

• Extended shelf life of 2 to 5 years without the use of preservatives;

• Storage and transport under standard conditions.

The method according to the invention ensures full utilization of the raw material - snails. It includes the following technological processes:

• Hatching of the snails in a specially constructed hatching room, • Rearing (fattening) of the snails in controlled conditions, as close as possible to their natural habitat, • Collection of caviar, mucus and hemocyanin under conditions causing minimal stress to the animals.

• Snail meat extraction • Snail shell processing.

• Preparation of protein and amino acid hydrolysis.

• Obtaining lyophilized products.

• Packaging and storage of the received products.

EXEMPLARY IMPLEMENTATION OF THE INVENTION

The present invention is illustrated by the following preferred exemplary embodiments, which in no way limit its scope.

Example 1.

Stage 1 Hatching of the snails in a specially constructed hatching room The reproduction of the snails takes place in artificial conditions. In specialized rooms, conditions close to those in nature are recreated. The animals are placed in special facilities /hammocks/. Here they fertilize and lay their eggs in a specific soil /peat/ placed in a bucket.

Rearing (fattening) of the snails in controlled conditions

The process of fattening the snails is carried out in specialized open-air farms. In them, the snails are intensively fed with combined starter feed - up to the first month and combined fattening feed - until they are ready for collection. The small snails created in artificial conditions are released at sunset in the outdoor farm. They are fattened on the farm for 3 to 4 months. During the fattening period and the period of extraction of the raw materials from the snails, a commercially available product LBB (Lactis Bacillus Bulgaricus) is added to the compound feed in some cases, which further stimulates the growth of the snails and acts favorably for the longer period of survival in hibernation.

Stage 2. Getting caviar from snails

After fertilization, the snails lay their eggs in a special soil (peat) placed in a bucket. Laying itself is a process that lasts from 46 to 48 hours. Then the bucket is emptied, the caviar is in the form of a ball, which is scattered in a container with a lid, where the snails hatch in a period of 12-15 days. According to the invention, up to 90% hatching of the snails is achieved, while in nature this process is only 10%. In the hatchery, on specially designed hammocks, the snails are placed at a density of 250-300 pieces per m . The animals are fertilized and lay their eggs (caviar) in specially placed polyethylene buckets filled with special soil (peat or similar soil). The snails are returned to the hammocks and the spawn is collected from the buckets. It is in the form of balls, each of which consists of about 100-120 individual caviar grains.

Stage 3 Obtaining slime from snails

Equivalent quantities of water and live snails in the case of 100 kg are placed in a 300 liter stainless steel vessel equipped with a low speed 60 rpm stirrer and 12 Volt low voltage pulse electrodes. With continuous stirring, the low-voltage voltage is applied to the vessel twice for 120 seconds. During periods of low voltage supply, snails become stressed and secrete mucus as a protective reaction. At the end of the process, the snails are removed from the container and returned to the hatchery. The solution of mucus and water was stored at a temperature of 4°C for 12 hours in cold rooms to homogenize it. The resulting slime solution was successively filtered on filters with a pore size of 50, 30, 20, 10, 5, 1 micron. This is followed by a process of additional filtration in order to concentrate, by means of reverse osmosis under standard conditions for the process.

Stage 4. Preparation of hemocyanin

Hemocyanin is extracted manually by an operator. For this purpose, the animal is taken in the hand and a slight S-shaped movement is performed along the lower part of the leg of the snail, ending at the end with a light sting. When boiling, up to two drops of a white thick liquid are released, which turns blue in contact with oxygen from the air.

Stage 5. Obtaining snail meat

According to the invention, the snails are subjected to heat treatment. To separate the meat from the shell, the snails are immersed in water preheated to 70°C for 30 seconds. The snail is immersed in pre-cooled water to 20°C in order to preserve the native form of the contained proteins, the meat is separated from the shell mechanically.

Stage 6. Preparation of protein hydrolyzate from meat

80 kg of freshly ground snail meat is added to 100 liters of preheated to 60°C, alkalized (pH=8) water in a stainless steel vessel with a volume of not less than 200 liters, equipped with a low-speed stirrer and a heater. A pre-calculated amount of Alcalase 2.5 enzyme, Novozymes, according to the manufacturer's instructions, is added to the mixture. The enzymatic hydrolysis process was carried out for 14 hours at 60°C and pH=8 of the reaction medium. The process is carried out under continuous pH control and, if necessary, corrected with a suitable alkaline solution. At the end of the process, the temperature of the reaction mixture was raised to 95°C and maintained for 4 hours. The reaction mixture is then cooled sharply to 20°C and acidified to pH=5.5 with a suitable acid.

Stage 7. Preparation of shell protein hydrolyzate

Snail shells obtained as described in step 5 were washed repeatedly with chilled water, then air-dried to constant weight. The dry shells are mechanically ground to a size of about 5 mm. 30 kg of freshly ground snail shells are added to 100 liters of pre-heated to 60°C alkalized pH=8 water in a 200-liter stainless steel vessel equipped with a low-speed stirrer and heater. A pre-calculated amount of Alcalase 2.5 enzyme, Novozymes, according to the manufacturer's instructions, is added to the mixture. The enzymatic hydrolysis process is carried out for 24 hours at 60°C and pH=8 of the reaction medium. The process is carried out under continuous pH control and, if necessary, corrected with an alkaline solution suitable for the purpose. At the end of the process, the temperature of the reaction mixture was raised to 95°C and maintained for 4 hours. The reaction mixture was then cooled sharply to 20°C and acidified to pH=4.5

The indicated quantitative results of the final products vary due to the fact that they depend on the raw material, which is a living organism and has individual indicators

Step 8 Preparation of lyophilized products by the method of step 2 to step 8

8.1. Obtaining lyophilized caviar

The resulting caviar is washed repeatedly with purified chilled water to remove traces of soil. After that, with the help of a blender (shaker), it is ground to obtain a homogeneous mixture. The pink colored mixture is frozen in a thin layer in the lyophilization trays at minus 40°C and the lyophilization process is carried out for 32 hours under controlled optimal conditions of vacuum and temperature.

Description of the product obtained by the method of stage 2

The product is a light, fine powder with a pinkish-reddish color. The product has a relatively low protein content of 25% and carbohydrates of 7%. The content of free fatty acids is below 1%.

Free amino acid content of lyophilized caviar

Amino acid content in mg/year minimally Maximum _ _ y Histidine 0.288 0.352 Arginine 0.243 0.297 Asparagine 0.162 0.198 Glutamine 0.252 0.308 4-Hydroxyproline 0.027 0.033 Serine 0.873 1,067 Aspartic acid 0.405 0.495 Glutamic acid 0.234 0.286 Threonine* 0.216 0.264 Glycine 0.27 0.33 Gamma-aminobutyric acid - - Tyrosine 0.072 0.088 Proline 0.117 0.143 _ _ y Methionine 0.018 0.022

valine" 0.171 0.209 41 Leucine/Isoleucine 0.207 0.253 tryptophan” 0.063 0.077 Phenylalanine" 0.126 0.154 Ornithine 0.576 0.704 Lysine" 0.261 0.319 General content 4,581 5,599

# essential amino acid

8.2. Preparation of lyophilized mucus

A concentrated mucus solution was frozen at -40°C in the lyophilization trays and the lyophilization process was conducted for 30 hours under controlled optimal vacuum and temperature conditions.

Description of the product obtained by the method of step 3

The product is a beige-brown fine powder. Biochemical analysis of the product shows that it is a complex mixture of proteins, carbohydrates, lipids, flavonoids, inorganic substances and low molecular weight compounds. The protein fraction is the main component of mucus in the amount of 40%.

Free amino acid content of lyophilized mucus

Amino acid content in mg/year Minimally maximum .... i Histidine 0.43 0.53 Arginine 0.11 0.13 Asparagine 0.39 0.47 Glutamine 0.40 0.48 4-Hydroxyproline 0.11 0.13 Serine 0.63 0.77

Aspartic acid 0.90 1.10 Glutamic acid 1.39 1.69 threonine* 0.43 0.53 Glycine 2.40 2.94 Gamma-aminobutyric acid - Tyrosine 0.05 0.06 Proline 0.45 0.55 Methionine 0.10 0.12 Valine* 1.64 2.00 Leucine/Isoleucine* 3.40 4.16 tryptophan" 0.19 0.23 Phenylalanine 0.82 1.00 Ornithine 1.48 1.80 Lysine* 1.73 2.11 General content 17.03 20.81

# essential amino acid

8.3. Preparation of lyophilized hemocyanin

The obtained hemocyanin is frozen in a thin layer in the lyophilization trays at minus 40-45°C and the lyophilization process is carried out for 36-38 hours, under controlled optimal conditions of vacuum and temperature.

Description of the product obtained by the method of step 4

Lyophilized hemocyanin is a fine white powder. The main components of the product are copper (Si) coordinated hemocyanin proteins, representing up to 90% of the product content.

8.4. Obtaining freeze-dried meat

The cooled meat is ground in a standard meat grinder, spread in a thin layer in the lyophilization trays, frozen at -45°C and the lyophilization process is carried out for 36 hours under controlled optimal conditions of vacuum and temperature.

Description of the product obtained from step 5

Freeze-dried meat is a brown powder with a high protein content of 75% and carbohydrates of 20% and a relatively low lipid level of less than 4%. Biochemical analysis shows that about 70% of the contained protein is water soluble.

Free amino acid content of freeze-dried snail meat

Amino acid content in mg/year minimally maximum Histidine 0.621 0.759 Arginine 0.18 0.22 Asparagine 0.333 0.407 Glutamine 2,106 2,574 4-Hydroxyproline 0.063 0.077 Serine 1,026 1,254 Aspartic acid 0.882 1,078 Glutamic acid 2,763 3,377 threonine* 0.549 0.671 Glycine 0.864 1,056 Gamma-aminobutyric acid - - Tyrosine 0.414 0.506 Proline 0.675 0.825 Methionine 0.135 0.165 Valine* 0.756 0.924

Leucine/Isoleucine 0.909 1,111 tryptophan* 0.162 0.198 Phenylalanine 0.603 0.737 Ornithine 7,803 9,537 Lysine* 2.16 2.64 General content 23,004 28,116

# essential amino acid

8.5. Preparation of lyophilized protein hydrolyzate from meat

After filtration (filter cloth), the obtained hydrolyzate is spread in a thin layer in the trays for lyophilization, frozen at -40°C and the lyophilization process is carried out for 34 hours, under controlled optimal conditions of vacuum and temperature.

Description of the product obtained from step 6

The product is a brown colored powder with a high water solubility of up to 90% on a dry matter basis. Main components of the lyophilized hydrolyzate are low molecular peptides 65% (up to 5 kD) and free amino acids. The product also contains protein, 10%, carbohydrates 10% and less than 4% lipids.

Free amino acid content of lyophilized snail meat hydrolyzate

Amino acid content in mg/year minimally maximum her Histidine 8.73 10.67 Arginine 2,043 2,497 Asparagine 14.76 18.04 Glutamine 30,429 37,191 4-Hydroxyproline 0.072 0.088 Serine 0.999 1,221 Aspartic acid 8,136 9,944

Glutamic acid 17,838 21,802 threonine* 0.405 0.495 Glycine 5,004 6,116 Gamma-aminobutyric acid Tyrosine 15,633 19,107 Proline 1,926 2,354 Methionine 6,084 7,436 Valine* 17,055 20,845 Leucine/Isoleucine* 25,713 31,427 tryptophan* 4,509 5,511 Phenylalanine 18,603 22,737 Ornithine 32,724 39,996 Lysine* 26,586 32,494 General content 237,249 289,971

# essential amino acid

8.6. Preparation of lyophilized shell protein hydrolyzate

The obtained hydrolyzate is spread in a thin layer in the trays for lyophilization, frozen at minus 40-45°C and the lyophilization process is carried out for 34-38 hours under controlled conditions of vacuum and temperature.

Description of the product obtained from step 7

The product is a beige-brown colored powder with a relatively low water solubility of up to 15% on a dry matter basis. The product dissolves well in dilute mineral acid solutions. Main organic components of the lyophilized hydrolyzate are up to 25% low molecular weight peptides and free amino acids.

Free amino acid content of lyophilized snail shell hydrolyzate

Amino acid content in mg/year minimally maximum Histidine 0.31 0.38 Arginine 0.02 0.02 Asparagine 0.18 0.22 Glutamine 2.99 3.65 4-Hydroxyproline 0.01 0.01 Serine 0.40 0.48 Aspartic acid 0.02 0.02 Glutamic acid 0.06 0.07 threonine* 0.23 0.28 Glycine 0.44 0.54 Gamma-aminobutyric acid - - Tyrosine 1.40 1.71 Proline 0.02 0.02 Methionine 0.06 0.07 Valine* 0.51 0.63 Leucine/Isoleucine* 0.46 0.57 tryptophan* 0.06 0.07 — y Phenylalanine 0.76 0.93 Ornithine 0.10 0.13 Lysine* 1.34 1.64 General content 9.37 11.45

# essential amino acid

Micronutrient content of lyophilized snail shell hydrolyzate

A trace element content in mg/year minimally maximum Calcium 189,010 231,020 Magnesium 172,800 211,200 Chrome 0.003 0.004 Sodium 5,020 6,140 Zinc 0.101 0.123 Iron 1,066 1,303

Example 2

Preparation of lyophilized caviar according to the invention in step 2

The resulting caviar is washed repeatedly with purified chilled water to remove traces of soil. After which, with the help of a blender (shaker), it is ground to obtain a homogeneous mixture. The pink colored mixture was frozen in a thin layer in the lyophilization trays at minus 45 °C and the lyophilization process was carried out for 38 hours under controlled optimal conditions of vacuum and temperature.

Description of the product:

The product is a light, fine powder with a pinkish-reddish color. The product has a protein content of 35% and carbohydrates 12%.

The remaining stages and the obtained products, their qualitative and quantitative characteristics are as in example 1.

Example 3

Preparation of mucus from snails, according to the invention in step 3

Equivalent amounts of water and live snails in the case of 100 kg are placed in a stainless steel container of a volume of not less than 250 liters equipped with a low-speed 60 rpm stirrer and electrodes for pulsed low-voltage voltage supply of 10 Volts. With continuous stirring, the low-voltage voltage is applied to the vessel for 120 seconds three times. During periods of low voltage supply, snails become stressed and secrete mucus as a protective reaction. At the end of the process, the snails are removed from the container and returned to the hatchery. The solution of mucus and water was stored at a temperature of 6 C for 10 hours in cold rooms to homogenize it. The resulting slime solution was successively filtered on filters with a pore size of 50, 30, 20, 10, 5, 1 micron. This is followed by a process of additional filtration in order to concentrate, by dialysis under standard conditions for the process.

The remaining stages and the obtained products, their qualitative and quantitative characteristics are as in example 1.

Example 3

Preparation of mucus from snails, according to the invention in step 3

Equivalent amounts of water and live snails in the case of 100 kg are placed in a stainless steel container with a volume of not less than 250 liters equipped with a low-speed 30 rpm stirrer and electrodes for pulsed low-voltage voltage supply of 12 Volts. With continuous stirring, a low-voltage voltage is supplied to the vessel for a short time of 60 seconds three times. During periods of low voltage supply, snails become stressed and secrete mucus as a protective reaction. At the end of the process, the snails are removed from the container and returned to the hatchery. The solution of mucus and water was stored at a temperature of 4 C for 14 hours in cold rooms to homogenize it. The resulting slime solution was successively filtered on filters with a pore size of 50, 30, 20, 10, 5, 1 micron. This is followed by a process of additional filtration in order to concentrate, through ultrafiltration under standard conditions.

Preparation of lyophilized mucus:

A concentrated mucus solution is frozen at a temperature of -45°C in the lyophilization trays and the lyophilization process is carried out for up to 38 hours under controlled optimal conditions of vacuum and temperature.

Description of the product:

The product is a beige-brown fine powder. Biochemical analysis of the product shows that it is a complex mixture of proteins, carbohydrates, lipids, flavonoids, inorganic substances and low molecular weight compounds. The protein fraction is the main component of the mucus and is in the amount of 43%.

The remaining stages and the obtained products, their qualitative and quantitative characteristics are as in example 1.

Example 4

Preparation of snail meat according to the invention in step 5

According to the invention, the snails are subjected to heat treatment. To separate the meat from the shell, the snails are immersed in water preheated to 70°C for 40 seconds. The snail is immersed in pre-cooled water to 20°C in order to preserve the native form of the contained proteins, the meat is separated from the shell mechanically.

Obtaining freeze-dried meat:

The cooled meat is ground in a standard meat grinder, spread in a thin layer in the lyophilization trays, frozen at minus 40°C and the lyophilization process is carried out for 30 hours under controlled optimal conditions of vacuum and temperature.

Description of the product:

Freeze-dried meat is a brown powder with a high protein content of 65% and carbohydrates of 14% and a relatively low lipid level of less than 4%. Biochemical analysis shows that about 70% of the contained protein is water soluble.

The remaining stages and the obtained products, their qualitative and quantitative characteristics are as in example 1.

Example 5

Preparation of protein hydrolyzate from meat, according to the invention in step 6

80 kg of freshly ground snail meat is added to 100 liters of preheated to 60°C, alkalized (pH=8) water in a stainless steel vessel with a volume of not less than 200 liters, equipped with a low-speed stirrer and a heater. A previously calculated (according to the manufacturer's instructions) amount of Alcalase 2.5 enzyme, Novozymes, is added to the mixture. The enzymatic hydrolysis process was carried out for 10 hours at 60°C and pH=8 of the reaction medium. The process is carried out under continuous pH control and, if necessary, corrected with a suitable alkaline solution. At the end of the process, the temperature of the reaction mixture was raised to 95°C and maintained for 4 hours. The reaction mixture is then cooled sharply to 20°C and acidified to pH=4.5 with a suitable acid.

Lyophilized protein hydrolyzate from meat:

After filtering with a filter cloth, the resulting hydrolyzate is spread in a thin layer in the trays for lyophilization, frozen at -45C° and the lyophilization process is carried out for 38 hours, under controlled optimal conditions of vacuum and temperature.

Description of the product:

The product is a brown colored powder with a high water solubility of up to 90% on a dry matter basis. Main components of the lyophilized hydrolyzate 75% are low molecular weight peptides up to 5 kD and free amino acids. The product also contains 15% protein, 14% carbohydrates and less than 4% lipids.

The remaining stages and the obtained products, their qualitative and quantitative characteristics are as in example 1.

Example 6

Preparation of shell protein hydrolyzate according to the invention in step 7

Snail shells obtained as described in step 5 were washed repeatedly with chilled water, then air-dried to constant weight. The dry shells are mechanically ground to a size of about 5 mm. In a stainless steel vessel with a volume of 200 liters equipped with a low-speed stirrer and a heater, 30 kg of freshly ground snail meat is added to 100 liters of preheated to 60°C alkalized pH=8 water. A pre-calculated amount of Alcalase 2.5 enzyme, Novozymes, according to the manufacturer's instructions, is added to the mixture. The enzymatic hydrolysis process was carried out for 20 hours at 60°C and pH=8 of the reaction medium. The process is carried out under continuous pH control and, if necessary, corrected with an alkaline solution suitable for the purpose. At the end of the process, the temperature of the reaction mixture was increased to 95 °C and maintained for 4 h. The reaction mixture was then cooled sharply to 20°C and acidified to pH=5.5.

The remaining stages and the obtained products, their qualitative and quantitative characteristics are as in example 1.

Claims (4)

1. A method for obtaining lyophilized products from snails including enzymatic hydrolysis and lyophilization, characterized in that the method is continuous and waste-free and includes the following stages: stage 1 hatching of snails and/or their fattening, stage 2 obtaining caviar from snails , step 3 obtaining snail mucus, step 4 obtaining snail hemocyanin, step 5 obtaining snail meat, step 6 obtaining protein hydrolyzate from snail meat, step 7 obtaining protein hydrolyzate from snail shell and step 8 process of lyophilization of the products from stage 2 to stage 7, with stage 1 being a process of hatching snails and/or fattening them in controlled conditions for a period of 3-4 months, in specialized premises and facilities, in stage 2 includes fertilization of the snails , spawning where the snails are located at a density of 250-300 pieces per m ' and the spawning process lasts from 46 to 48 hours, stage 3 the process of obtaining slime from snails takes place in a vessel with equi valent quantities of water and live snails, the vessel is equipped with a stirrer of 30 to 60 revolutions per minute and electrodes for the pulsed supply of low-voltage voltage from 10 to 12 Volts, which is supplied for a time of 60-120 seconds from 2 to 3 times, the snails are removed from the vessel, the solution is homogenized for 10 to 14 hours at a temperature of 4-6°C, it is successively filtered through filters with a pore size of 50, 30, 20, 10, 5, 1 microns, the resulting filtrate is subjected to additional filtration, stage 4 the process of obtaining hemocyanin is carried out manually, making an S-shaped movement along the lower part of the leg of the snail, ending at the end with a light sting and releasing the hemocyanin, stage 5 process of obtaining meat from snails, which is carried out by heat treatment, separating the meat from the shell in water pre-heated to 70°C for 30-40 seconds, followed by mechanical removal of the meat and immersion in water pre-cooled to 20°C, stage 6 process on obtaining a protein hydrolyzate from snail meat, which is carried out by enzymatic hydrolysis in a stainless steel vessel equipped with a low-speed stirrer and a heater in alkalized (pH = 8) water, which is preheated to 60°C, the ground snail meat is added and an enzyme is added, the process takes place for 10-14 hours at a temperature of 60°C to 95°C with continuous pH control and, if necessary, corrected with an alkaline solution, followed by cooling to 20°C and acidification to pH=4.5-5.5, step 7 the process of obtaining snail shell protein hydrolyzate proceeds as the snail shells obtained from step 5 are washed repeatedly with chilled water, then air-dried to a constant weight, followed by mechanical grinding to a size of 5 mm, placed in a stainless steel vessel with a low-speed stirrer and a heater to preheated to 60°C alkalized pH = 8 water, an enzyme is added to the mixture and the enzymatic hydrolysis process is carried out for 20 to 24 hours at 60°C and pH=8 of the reaction medium, the process takes place under continuous pH control and if necessary corrected with an alkaline solution, at the end of the process, the temperature of the reaction mixture is increased to 95°C and maintained for 4 hours, followed by sharp cooling to 20°C, after which it is acidified to pH=4.5-5.5, stage 8 the products obtained from stages 2, 3, 4, 5, 6 and 7 are lyophilized. 2. Method according to claim 1, characterized in that in a given case step 2 and step 3 are repeated multiple times. 3. Method according to claim 1, characterized in that the lyophilization of the products takes place at a temperature of minus 40-45°C for a time of 32 to 48 hours. 4. Method according to claim 1, characterized in that in step 3 the additional filtration is carried out by reverse osmosis or dialysis or ultrafiltration. 5. Products obtained by the method according to claim 1, characterized in that it is lyophilized caviar, which is a fine powder with a pinkish-reddish color, with a protein content of 25-35%, carbohydrates 7-12% and a content of free amino acids as follows: Amino acid content in mg/g minimum maximum Histidine 0.288 0.352 Arginine 0.243 0.297 Asparagine 0.162 0.198 Glutamine 0.252 0.308 4-Hydroxyproline 0.027 0.033 Serine 0.873 1,067 Aspartic acid 0.405 0.495 Glutamic acid 0.234 0.286 Threonine 0.216 0.264 Glycine 0.27 0.33 Gamma-aminobutyric acid - - Tyrosine 0.072 0.088 Proline 0.117 0.143 Methionine 0.018 0.022 Valin 0.171 0.209 Leucine/Isoleucine 0.207 0.253 Tryptophan 0.063 0.077 Phenylalanine 0.126 0.154 Ornithine 0.576 0.704 Lysine 0.261 0.319 General content 4,581 5,599 6. Products obtained by the method according to claim 1, characterized in that it is a lyophilized slime, which is a fine powder with a beige-brown color, the protein fraction is 40 - 45% and it has a content of free amino acids as follows: Amino acid content in mg/g minimum maximum Histidine 0.43 0.53 Arginine 0.11 0.13 Asparagine 0.39 0.47 Glutamine 0.40 0.48 4-Hydroxyproline 0.11 0.13 Series 0.63 0.77 Aspartic acid 0.90 1.10 Glutamic acid 1.39 1.69 Threonine 0.43 0.53 Glycine 2.40 2.94 Gamma-aminobutyric acid - - Tyrosine 0.05 0.06 Proline 0.45 0.55 Methionine 0.10 0.12 Wales 1.64 2.00 Leucine/Isoleucine 3.40 4.16 Tryptophan 0.19 0.23 Phenylalanine 0.82 1.00 Ornithine 1.48 1.80 Lysine 1.73 2.11 General content 17.03 20.81 7. Products obtained by the method according to claim 1, characterized in that it is lyophilized hemocyanin, which is a fine white powder containing copper (Si) coordinated hemocyanin proteins, 8. Products obtained by the method according to claim 1, characterized in that it is lyophilized snail meat, which is brown in color powder with a protein content of 65-75%, carbohydrates 14-20% and with a content of free amino acids, as follows: Amino acid content in mg/g minimum maximum Histidine 0.621 0.759 Arginine 0.18 0.22 Asparagine 0.333 0.407 Glutamine 2,106 2,574 4-Hydroxyproline 0.063 0.077 Serine 1,026 1,254 Aspartic acid 0.882 1,078 Glutamic acid 2,763 3,377 Threonine 0.549 0.671 Glycine 0.864 1,056 Gamma-aminobutyric acid - - Tyrosine 0.414 0.506 Proline 0.675 0.825 Methionine 0.135 0.165 Wales 0.756 0.924 Leucine/Isoleucine 0.909 1,111 Tryptophan 0.162 0.198 Phenylalanine 0.603 0.737 Ornithine 7,803 9,537 Lysine 2.16 2.64 General content 23,004 28,116 9. Products obtained by the method according to claim 1, characterized in that it is a lyophilized protein hydrolyzate of snail meat, which is beige-brown in color powder, contains low molecular weight peptides 65-75%, protein 10-15 %, carbohydrates 1014 % and amino acids as follows: Amino acid content in mg/g minimum maximum Histidine 8.73 10.67 Arginine 2,043 2,497 Asparagine 14.76 18.04 Glutamine 30,429 37,191 4-Hydroxyproline 0.072 0.088 Serine 0.999 1,221 Aspartic acid 8,136 9,944 Glutamic acid 17,838 21,802 Threonine 0.405 0.495 Glycine 5,004 6,116 Gamma-aminobutyric acid Tyrosine 15,633 19,107 Proline 1,926 2,354 Methionine 6,084 7,436 Valin 17,055 20,845 Leucine/Isoleucine 25,713 31,427 Tryptophan 4,509 5,511 Phenylalanine 18,603 22,737 Ornithine 32,724 39,996 Lysine 26,586 32,494 General content 237,249 289,971 10. Products obtained by the method according to claim 1, characterized in that it is a lyophilized snail shell hydrolyzate, which is a beige-brown powder in color and has a content of free amino acids and trace elements as follows: Amino acid content in mg/g minimum maximum Histidine 0.31 0.38 Arginine 0.02 0.02 Asparagine 0.18 0.22 Glutamine 2.99 3.65 4-Hydroxyproline 0.01 0.01 Serine 0.40 0.48 Aspartic acid 0.02 0.02 Glutamic acid 0.06 0.07 Threonine 0.23 0.28 Glycine 0.44 0.54 Gamma-aminobutyric acid - - Tyrosine 1.40 1.71 Proline 0.02 0.02 Methionine 0.06 0.07 Wales 0.51 0.63 Leucine/Isoleucine 0.46 0.57 Tryptophan 0.06 0.07 Phenylalanine 0.76 0.93 Ornithine 0.10 0.13 Lysine 1.34 1.64 General content 9.37 11.45 A trace element content in mg/year minimally maximum Calcium 189,010 231,020 Magnesium 172,800 211,200 Chrome 0.003 0.004 Sodium 5,020 6,140 Zinc 0.101 0.123 Iron 1,066 1,303