RU2556121C1 - Tea fungus infusion production method - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: method envisages Medusomyces Gisevii Lindau microorganism cultivation under aeration conditions at a temperature of 23-25°C in the carbohydrate-containing medium; the carbohydrate source and growth promoter are represented by dried homogenate of dead bees in an amount of 8-10 g per 1 l of the nutrient medium.

EFFECT: invention allows to accelerate the end product maturation and producer-microorganism biomass accumulation as well as improve the manufactured product taste properties.

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Description

The invention relates to the field of biotechnology, microbiological, food and processing industries and can be used to obtain kombucha infusion by growing it in an aqueous solution containing a beekeeping product - bee morbidity.

Bee killing is a complex of dead bees. From a technological point of view, scorching is a natural raw material that contains protein, chitin, melanins, wax, vitamins, mineral salts, organic acids, hydrocarbons, flavonoids, biologically active substances that stimulate cell growth and development, free amino acids, fatty acids, nucleic acids. Fundamentally, bee mortem contains all beekeeping products (honey, bee bread, pollen, wax, bee venom, propolis, royal jelly) that contain more than 400 different components (Y. Konstantinov. Beekeeping products. Natural medicines. - M.: Publishing House in “Centerpolygraph.” - 2014, 160 s). It is enriched with wax and wax-like substances containing esters, saturated hydrocarbons, free fatty acids, aromatic substances.

The number of flavonoid and phenolic compounds (14-16%) characterizes the so-called "colored alcohols" of carotenoid nature and is used as a determining indicator of preparations obtained from bees. Noteworthy is the abnormally high value of the content of vitamin C - 0.5-0.7 mg per 1 g of bee extract. For comparison: the vitamin C content in citrus fruits is 0.4-0.6 mg per 1 g. Ascorbic acid, which is present in bee subpestilence, is of great importance for its use in medical and preventive practice.

The most interesting from the point of view of modern biotechnology is chitin. Chitin is an acetylated aminopolysaccharide that is part of the cuticle of insects, the shell of crustaceans and the cell wall of fungi. In the cuticle of insects, including bees, chitin is in combination with melanins. Moreover, the ability of chitin under certain conditions (under the influence of enzymes, organic acids) to turn into a biologically active substance - chitosan, used in medicine, food industry, cosmetics, agriculture, radiobiology, toxicology, pharmacology, immunology and veterinary medicine is important. Today, there are more than 70 known areas of use of chitosan (Chitin and chitosan. Obtaining, properties and application / Ed. By K.G. Skryabin, G.A. Vikhoreva, V.P. Varlamova. - M .: Nauka, 2002. - 368 p.).

Thus, bee killing is an extremely complex complex containing biologically active substances that stimulate the growth and development of cells of plant, animal and microbial origin, enhancing their metabolism and their production of vital biologically active substances used both in medicine and in food industry (Nemtsov S.V., Zueva O.Yu., Khismatullin R.G. and others. Chitosan from death - a new product of bees / Beekeeping. - 2001. - No. 5. - S. 50-51; Nemtsov S.V. ., Zueva O.Yu., Khismatullin R.G. Apizan / http: // be ewellms.ru/ru / tentorium - shop - apizan).

A known method of growing Kombucha in a 10% solution of sugar and tea (Khachatryan V. Kombucha. - St. Petersburg: Dilya, 2007, 160 C.).

The disadvantage of this method is the long ripening (up to 8 days) of the infusion and a significant consumption of a valuable food product - sugar.

There are also known methods of obtaining infusion of Kombucha by cultivating it in aqueous solutions containing nitrogen-containing substances (ammonium salts, ammonium phosphates, citric acid, starch, glycerin, yeast extracts (Danielyan L.T. Kombucha and its biological properties. - M .: Medical literature, 2010. - 83 p.).

The disadvantage of this method is the high cost of the starting products and the weak stimulation of symbionts (yeast and acetic acid bacteria that make up the basis of Kombucha). At the same time, there is no acceleration in the maturation of the tea drink and an improvement in the taste of the products.

A known method of growing yeast on mineral nutrient media containing sources of carbon, nitrogen, phosphorus, trace elements using as a stimulator of yeast growth α-aminocarbolic acids that are part of natural components such as arginine, leucine and lysine, as well as B vitamins, especially pantothenic acid and pyridoxine (French Patent No. 1538957, CL C12C, 1969).

The disadvantage of this method is the high cost of the starting products.

The closest analogue of the proposed method is a method for producing yeast biomass under aeration conditions at a temperature of 25 + 40 ° C and pH 3.0 + 6.0 on a nutrient medium containing sources of carbon, nitrogen, mineral additives and growth stimulants. As a nutrient medium, hydrolysates or fermentolysates of grain raw materials and / or waste from flour milling are used. As biostimulants of yeast cell growth, they use (all 100%): sodium sulfite (1 · 10 ⁴ ± 1 · 10 ^-8 ), gibberillic acids (1 · 10 ^-7 ± 1 · 10 ^-17 ), water-soluble chitin oligosaccharines (1 · 10 ^-5 ± 1 · 10 ^-9 ), with a total concentration (1 · 10 ^-5 ± 1 · 10 ^-10 ) to the volume (RF Patent No. 2384612, class C2, 2007).

The disadvantage of this method is that the stimulating effect of increasing the biomass of yeast is achieved only when combined with these components, which makes the method very expensive and economically inefficient. Moreover, the biomass increase is only 5-15%, which is clearly insufficient, since there is a direct correlation between the biomass and the final product produced by the producer microbe, because the larger the biomass, the greater the final product produced. In addition, the method involves the use of a growth promoter of water-soluble oligosaccharides of chitin (chitosan), the preparation of which is technologically difficult, time-consuming and expensive.

Since Kombucha during the growing process synthesizes and produces acetic, succinic, gluconic, oxalic, citric, malic, pyruvic acid and enzymes: catalase, lipase, protease and carboxylase in the culture fluid, the chitin contained in the bees under the influence of these acids undergoes natural succination, decarboxylation and deacylation, turning into an easily accessible and high-quality nutrient substance for microorganism cells - producer of Kombucha drink - hydrophilic Ationic biopolymer - chitosan, which is a 2-amino-duoxy-b-D-glucan aminopolysaccharide, which is a highly active stimulator of cell growth and development, as well as an ideal enhancer of taste and aroma (L. S. Galbraich. Chitin and chitosan. Structure, properties, application. // Soros Educational Journal. - 2001. - T. 7. - No. 1. - S. 51-56).

The use of chitin-containing raw materials - bee killing as food for Kombucha in order to obtain a drink allows replacing an expensive food product in the growth medium - sugar with a cheaper, more affordable and more active food source - a waste beekeeping product containing a number of biologically active components for full development, synthesis and production of metabolites useful for humans, a number of which (acetic, oxalic, succinic acids) ferment the polysaccharide into monosaccharides, and enzymes (amylase, decarboxy basics, succinate dehydrogenase) convert chitin of bee subpestilence to chitosan - the most valuable biologically active substance, thereby eliminating the need to obtain chitosan in an industrial way, which is very expensive, complex, and most importantly - technologically dangerous, since it is necessary to convert chitin to chitosan by deacylation and demineralization of raw materials apply concentrated acids, alkalis (50-80%), as well as high temperatures (130-180 ° C).

The objective of the invention is to obtain the infusion of tea mushroom, which allows to accelerate the ripening of the infusion, improve the taste of the finished product, accelerate the growth and increase the biomass of the microorganism - producer and reduce the cost of obtaining the final product by replacing an expensive power source - sugar with a more affordable and cheaper power source - a waste product of beekeeping - dead bees.

The problem is solved in that the method of obtaining infusion of Kombucha involves cultivating the microorganism under aeration conditions at room temperature (23-25 ° C) in a carbohydrate-containing nutrient medium, as a source of carbohydrates and a growth stimulator, dried bee homogenate is used at a rate of 8-10 g per 1 liter of culture medium.

The microbial association Medusomyces Gisevii Lindau, which is a symbiosis of strains of Sacharomyces cerevisae, Torula, Acetobacter xilinum, is used as a microorganism producing infusion of Kombucha.

As sources of carbohydrates and a growth stimulant, beekeeping wastes are used - dead bees containing high molecular weight compounds - polysaccharides as an integral part of chitin.

Replacing sugar with a chitin-containing component, which is the death of bees in the culture medium of Kombucha cultivation, significantly accelerates the processes of cell metabolism due to the conversion of chitin into chitosan in the culture medium, which has metabolism-stimulating and growth-promoting effects, as well as giving the taste and aroma to the drink.

At the first stage of obtaining kombucha infusion, the main technological raw material is prepared - bee subpestilence. For this purpose, fresh dead bees are collected without signs of mold and decay. Raw materials are sieved through a sieve with large cells to separate it from small debris. Then the death is dried in an oven, oven at a temperature of + 40-45 ° C or above a central heating battery. The dry material is homogenized in a ball mill or electric grinder. The degree of dispersion is controlled microscopically, which should be in the range of 60-90 microns. The resulting powder is packaged in paper bags of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50 g, sterilized by autoclaving at 100 ° C and a pressure of 1.0-1.5 atmospheres. Sterilized bags with homogenate (powder) of dead bees are stored in the refrigerator and used as carbon sources in the culture medium.

The technological process of obtaining infusion of tea mushroom is as follows. To prepare the infusion of kombucha use a sterilized three-liter jar with a wide neck, which is covered with several layers of gauze. The lower layer is separated from the mother fungus and put it in a jar of boiled water (1.5-2 l). First, the sterilized bee homogenate (powder) prepared at the first stage is put on the bottom of the can, which is poured into a sterile gauze bag, tied with a thread and lowered to the bottom of the can, the ends of the string are left outside the neck of the can to replace the nutrient material after draining the finished infusion and making another portion nutrient material (dried homogenate of dead bees).

The jar with the contents is placed in a warm (20-25 ° C) place away from the window to prevent direct sunlight. At the end of the cycle (depending on the degree of maturation of the infusion after 2, 3, 4, 5, 6, 7, 8, 9 and 10 days), the infusion of the fungus is drained through a layer of gauze, and a control tasting is carried out. As it ripens, it is used as a drink.

The degree of ripening of the drink is evaluated by tasting (by taste) and chemically by the concentration of glucuronic acid, which is not detected as it ripens, as well as by the accumulation of Kombucha biomass during cultivation.

For experimental verification of the proposed method, samples of a drink prepared by cultivating kombucha for 2, 3, 4, 5, 6, 7, 8, 9, 10 days after introducing various amounts of bee homogenate into the nutrient medium (1; 2; 4; 8; 10 and 20 g) per 1 liter of culture medium, which was 0.1; 0.2; 0.4; 0.8-1.0; 2.0 wt. %

In parallel, control samples of the tea mushroom drink were obtained, grown for 2, 3, 4, 5, 6, 7, 8, 9, and 10 days according to the known method on a nutrient medium containing 100 g of sugar and 5 g of black tea per 1.0 l of culture medium.

Example 1. Obtaining samples of Kombucha drink by cultivating a producer microorganism for 2 days when keeping bee homogenate in the nutrient medium in an amount of 0.1; 0.2; 0.4; 0.8; 1.0; 2.0 wt. %

Example 2. While maintaining the conditions of example 1, get samples of the tea mushroom drink by growing the producer for 3 days when the content in the nutrient medium of similar concentrations of bee homogenate shown in example 1.

Example 3. Get samples of the drink of tea mushroom by culturing the producer for 4 days while maintaining the conditions of example 1.

Example 4. Get samples of the drink of Kombucha by growing the producer for 5 days on media containing similar concentrations of bee homogenate, shown in example 1.

Example 5. Get samples of the drink of Kombucha by growing the producer for 6 days while maintaining the conditions of example 1.

Example 6. Receive samples of the drink of Kombucha by growing the producer for 7 days while maintaining the conditions of example 1.

Example 7. Get samples of the drink of Kombucha by growing the producer for 8 days while maintaining the conditions of example 1.

Example 8. Receive samples of the drink of Kombucha by growing the producer for 9 days while maintaining the conditions of example 1.

Example 9. Get samples of the drink of Kombucha by growing the producer for 10 days while maintaining the conditions of example 1.

Technological indicators for obtaining a tea mushroom drink are presented in table 1.

Thus, the introduction of the homogenate of dead bees into the culture medium of Kombucha cultivation leads to a significant improvement in all technological parameters of the quality of the synthesized product - Kombucha drink: the growing time (maturation of the drink) is reduced by 50%; biomass increase increases by 59%; the expenditure coefficient for the nutrient substrate (power source) is reduced by 10 times (1 mg / ml of bee death versus 10 mg / ml of sugar in the known method).

The invention in comparison with the known technical solution has the following advantages:

- acceleration of maturation of the final product;

- the use of cheap waste beekeeping product as a food source;

- saving valuable foods: tea and sugar;

- acceleration of biomass accumulation of the producer microorganism;

- improving the taste of the resulting product; the conversion of chitin of bee subpestilence to chitosan due to production by the producer during the cultivation of organic acids and enzymes;

- low cost, environmental friendliness and higher biological usefulness of the food product.

Claims (1)

A method of obtaining kombucha infusion, including cultivating the microorganism Medusomyces Gisevii Lindau under aeration conditions at 23-25 ° C in a carbohydrate-containing medium, characterized in that a dried bee subpesticide homogenate is used as a source of carbohydrates and growth stimulator at the rate of 8-10 g per 1 l nutrient medium.