Producer melaninogenica biologically active additives "stromelysin"

 

(57) Abstract:

Mutant strain 1-365 Antarctic black yeast Nadsoniellanigravar. hesuelica recommended as a producer melaninogenica biologically active additives. The specified strain produces a wide range of biological macrobiotic activity. Protivopedikuleznoe action supplements stop the ravages of unstable free radicals. Apply an additive for foods, drugs and cosmetics. The additive is non-toxic, is a fine black crystalline powder with a shimmering glitter or its aqueous solution. 3 C.p. f-crystals.

The invention relates to biotechnology and can be used in food, cosmetic and pharmaceutical industry for the production and application of biologically active melaninogenica treatment-and-prophylactic food supplements for diet, additives in cosmetics as a highly active components protective or curative-restorative values, as well as additives in medicines to enhance or modify their therapeutic action.

It is known that based on the use of biomass and other mikroorganismen areas of action. Proactive they are mainly polysaccharides, carotenoids, peptides, vitamins, enzymes (W. A. Heyes , S. H. Wright Edible Mushrooms. In: Economic microbiology, vol. 7, n 4, ed. dy Rose A. H., London, Academ Press, 1979, p. 141.)

Known enamelin (grape melanin) that is proposed for use in the food industry as a food additive to soft drinks [1].

Known melanin biosynthetic for the cosmetic industry company [2].

These products have been adopted for the prototype the purpose of the final product, i.e., for use with similar objectives, namely as a food additive and additive in cosmetics, respectively.

The advantages and technical effects of the invention consist in identifying the source of effective biological additives, with new, not previously known properties not inherent in the known means of similar purpose, namely in the application of the known Antarctic mutant strain 1-365 black yeast Nadsoniella nigra var. hesuelica (Lyakh, C. P., Ruban E. L. Types of pigment mutants Nadsoniella nigra and efficiency of RadioButton. - Microbiology, 1969, T. 38, S. 663.; Ruban E. L., Lyakh, C. P., Volkov, I. M. Mutant Nadsoniella nigra, synthesizing extracellular supplements "Stromelysin" a wide spectrum of action and potential use. Discovered the useful properties of Astromania" strain 1-365 were not previously known and is not described in the literature, although the properties of the melanin - chemical and physico-chemical us was radiated (Ruban E. L., Lyakh, C. P., khrulyova I. M., Titov, I. A. "Melanin pigments Nadsoniella nigra" Izvestiya an SSSR, ser. Biol., 1969, No. 1, S. 134.).

It is known that microbial physiology and are a unique group of substances (Lyakh, C. P., Ruban E. L. Microbial physiology. M.: Nauka, 1972; Lyakh, C. P. Microbial melaninogenica and its functions. M.: Nauka, 1981) and that some of them are on the level or range of activity is significantly superior to vegetable, animal, synthetic and semi-synthetic (biosynthetic) melanin. Melanin biosynthetic at the above patent is obtained from the individual chemical substances with the help of the enzyme preparation, and as substrates using such expensive reagents, as indole, l-DOPA, tyramine and tyrosine, the production of which is very limited. Established industrial production of the claimed supplements are cheaper and can be scaled indefinitely. In addition, it can be arranged in any place, even in Antarctica, because it does not depend on proximity or dostupnaja Nadsoniella nigra var. hesuelica, which is the producer of melaninogenica anticancer drug [3].

The Antarctic flora, delivered in extreme natural conditions, have developed unique mechanisms and substances, ensuring the survival and active life in such a harsh environment as Antarctica (Lyakh, C. P. Adaptation of microorganisms to low temperatures. M.: Nauka, 1976). These substances and microorganisms are of great interest for the pharmaceutical industry, so forward-thinking professionals rightly believe Antarctic flora pharmaceutical Klondike ("Bacterial biomass Antarctica was pharmaceutical Klondike" - Financial news, N 95 329 8 Oct. 1996, according to Agency Reuters).

Antarctic roots mutant strain 1-365 Nadsoniella nigra var hesuelica gave him not only a high resistance to adverse conditions of its historical homeland in their extreme expression, but also great potential persistent resistance to the effects of many other fatal factors. The genetic apparatus of cells of strain is also well protected, and even mutant strain was obtained under the action of very high doses RNO to associate with the existence in their cell walls, on the border with the outside world, the black pigment melanin, stable free radicals at a high concentration (Ruban E. L., Fomin, C., Master, S. P. a Study of melanin Nadsoniella nigra by the EPR method. - DAN SSSR, 1968, I. 182, No. 5, S. 1219). This black pigment extracted from biomass, and is the basis for anticancer drug "Stromelysin".

However, as it turned out, in addition to its main purpose, "Stromelysin" strain 1-365 exhibits a broad spectrum of biological macrobiotic activity, the boundaries of which are not yet clear.

The essence and purpose of the invention is the use of strain 1-365 Antarctic black yeast Nadsoniella nigra var. hesuelica as unlimited and easily reproducible in the production source of microbial melanin used for a new purpose, namely as a highly active start melaninogenica biologically active additives that may be used in food, cosmetic and pharmaceutical industries. Shared between all of these forms of supplements is primarily protivopedikuleznoe action, stopping the destructive action of unstable free radicals. The basis for this effect is provided by the system melanin Nadsoniella nigra var. hesuelica were the first of microbial melanin, which found subtropically properties.

Based on the foregoing, Stromelysin" as treatment-and-prophylactic food additive can and should be used primarily in a special diet for the correction of the violations underlying the free radical pathologies: for the prevention and treatment of recurrences and metastases in cancer patients after radical treatment for the prevention of malignant tumors with borderline forms of the disease and with the same purpose in families with a high cancer risk, receiving a real threat; to prevent disorders in people who are in conditions of high radioactive background of long-term or short-term (in the latter case, for example, "Stromelysin" should be added to drinks for the flight personnel and passengers); for the removal of radionuclides and radiation sickness. "Stromelysin" in the form of a dietary Supplement also helps with stomach ulcers and 12 duodenal ulcers (ulcers fast subcourse; when XP. gastritis quickly disappear all the typical symptoms, "Stromelysin" restores the structure of the liver and pancreas, activates the immune system "cleans" diagenic situations, in combination with lotions successfully cures piles, relieves symptoms of XP. prostatitis with restoration of natural features and other

Stromelysin it is preferable to add a spring or mineral water or soft drinks, cocktails, or any other acceptable food product. In acid environments, water-soluble form of the additive may precipitate a brown flakes that does not detract from its biologically active virtues.

For creams, lotions, pastes and other cosmetic additive protects skin from the same unstable free radicals and UV rays, especially the hard ultraviolet highlands, the Antarctic summer and UV radiation technogenic character. Photoprotection effect of melanin strain 1-365 previously been shown in experiments on irradiation of UV light germicidal lamps are sensitive to UV light of organisms, which is very important, in the presence of melanin formation cytotoxic substances under the action of UV light was observed. The pigment is protected cells only at the moment of exposure (Lyakh, C. P., Ruban E. L. Photoprotection effect pigment Nadsoniekka nigra - Microbiology, 1968, T. 37, vol. 5, S. 862). Part of the mechanism of which is unclear.

The additive may be introduced into food or cosmetic, or therapeutic agent as an individual substance or combinations thereof (compositions) with other substances, including biologically active, and also be entered as a component in other supplements.

The additive is non-toxic even at high concentrations (observations on the Order of C. P. and experiments on the test-object of the ciliate Tetrahymena pyriformis - GOST 28178-89 and mice). It was proved the absence of acute toxicity in biomass Nadsoniella nigra var. hesuelica 1-365 (experiments on the test ciliates). Thus, working with culture and with the addition of (biotech part at the stage of its preparation) safe for staff, as a safe additive itself.

Getting the dietary Supplement.

Mutant strain 1-365 Antarctic black yeast Nadsoniella nigra var. hesuelica as producer supplements "Stromelysin" cultivated in a known manner under conditions of aeration and agitation in a liquid nutrient medium, for example, the following composition: g/l : K2HPO40,1; CaCl20,03; MgSO40,05; NaCl, 0,05 with glucose or sucrose (1.5 to 2%), liquid yeast autolysate (1%) pH 3.5 - 3 26 - 28oC during the week. To increase biomass yield and STABILO to grow biomass producer strain 1-365 can and apparatus for aeration and mixing, if the cultivation is carried out in a thin layer of fluid with a good enough natural aeration. The biomass from the liquid medium is separated by centrifugation and washed repeatedly with water, then autoclave at 0.5 ATM with 0.5 N. NaOH for 1 - 2 h-Cell mass is removed by centrifugation, centrifugal acidified with conc. HCl to pH 2. Dried out. So get a water-insoluble additive. To obtain water-soluble forms the washed precipitate slightly dry or immediately dissolved in a small amount of 12 - 25% ammonia solution, followed by a soft evaporation of unreacted ammonia and water. So get ammonium salt. Na+and K+-salt is obtained by dissolving the precipitate in 0.5 N. NaOH or KOH, respectively dialysis in distilled water and drying.

Ready-made additive "Stromelysin" represents black crystals with a bright shimmering Shine or concentrated solutions in water. For convenience of packing the original crystals RUB. The powdered form of the final product makes it easy to dispense the drug. Aqueous solutions of additives are color from cognac to black depending on the concentration. Concentrated solutions are stable and also the"Astromania" strain 1-365 on the test-volunteers as a dietary Supplement:

Example 1. Patient N. L., 1938 born. Data endoscopy: from 22.09.95 aggravation of ulcers bulbs 12 duodenal ulcer with a diameter of 0.5 - 0.6 see Supplement "Astromania in drinking water is 10 mg 3 times a day for 14 days. 20.10.95. : healed ulcer (white scar); 28.01.97. the bulb 12 duodenal ulcer - without a pathology.

Example 2. Patient I., 1932 born., physician, melanoma of the skin of the anterior chest wall. 24.06.94 excision education. Histology: nodular form, level IV invasion 8 mm. Adding "Astromania" drink with 25.06.94. first, in doses of 60 to 30 mg / day with subsequent dose reduction. Since the beginning of October and now 2.5 - 3 mg / day breaks to drink. May 1997: without recurrence, metastasis no.

Example 3. Pilot C., 1912 born. All possible senile Algie, adding "Astromania" in "mixture diet was significantly accelerated and increased analgesic effect.

Example 4. Test B. , 1938 born. Adding "Astromania" balm "gold star" has had a noticeable effect in the treatment of runny nose, previous use of this tool for this purpose without additives were ineffective. The additive into the rinse and drink quickly removed sore throat.

Example 5. Test B., 1938 born. January 96, it is time to Relax. ezidis in December 1996, reuse of Astromania" as additives in drinking water "Holy spring" - the symptoms completely disappeared after a month.

Example 6. Volunteer L., 1936 born. The introduction of Astromania" in the vaseline defended the skin of the hands from burns under a UV lamp . The other hand only with vaseline at the same exposure was badly burned.

Thus, from comparison with known means the same destination can be seen that the mutant strain 1-365 Antarctic black yeast Nadsoniella nigra var. hesuelica is a producer of valuable high-level and non-toxic additives for food products, cosmetics and medicines.

1. The use of a mutant strain 1-365 Antarctic black yeast Nadsoniella nigra var. hesuelica as producer melaninogenica biologically active additives.

2. The use of a mutant strain 1-365 Antarctic black yeast Nadsoniella nigra var. hesuelica as producer melaninogenica biologically active additives under item 1 for food.

3. The use of a mutant strain 1-365 Antarctic black yeast Nadsoniella nigra var. hesuelica as producer melaninogenica biologically active additives on p. 1 space for the hesuelica as producer melaninogenica biologically active additives under item 1 for medicines.

 

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