Producer melaninogenica biologically active additives "stromelysin"

 

(57) Abstract:

Natural strain 365 Antarctic black yeast Nadsoniella nigravar. 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, cosmetics and medicines. The additive is non-toxic, is a fine black crystalline powder with shimmering Shine and its aqueous solution. 1 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 for amplification or modification of the remedial 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 and other (Hayes W. A. , Wright S. H. Edible mushrooms. In: Economic microbiology logy, vol. 4, ed. by Rose A. H., London, Acad. Press, 1979, p. 141).

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

Known melanin biosynthetic for cosmetics company L OERAL (Paris) (FR 269 8336-A1, C 12 P 1/00, 1994).

These products, namely emmalani and melanin biosynthetic adopted for the prototype the purpose of the final product, i.e., for use with similar objectives, namely as a dietary Supplement is the first and additives in cosmetics is the second.

The advantages and technical effects of the invention consists in identifying the source of effective biological additives, with new, not previously known properties not inherent to the known properties of similar purpose, namely in the application of the known Antarctic strain 365 black yeast Nadsoniella nogrs var. hesuelica (Lyakh, C. P., Ruban E. L. Antarctic black yeast Nadsoniella nogrs var. hesuelica. Izvestiya an SSSR, ser. Biol., 1970, No. 4, S. 581) as is ncial use. Discovered the useful properties of Astromania were not previously known and is not described in the literature.

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 level or spectrum 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 unlimited mashtabirovanie. In addition, it can be arranged in any place, even in Antarctica, because it does not depend on proximity or availability of sources of raw materials, as, for example, enamelin.

A known strain 365 Antarctic black yeast Nadsoniella nigra var. hesuelica, which is the producer of melaninogenica anticancer drug [performance communications mechanisms and substances providing and 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 from 8 Oct. 1996, according to Reuters.).

Antarctic origin Nadsoniella higra var. hesuelica strain 365 has given her not only a high resistance to adverse conditions of its historical homeland in their extreme expression, but also great potential persistent resistance to impact many other fatal factors. The genetic apparatus of cells of strain is also well protected (Lyakh, C. P. Melanopogon Antarctic black yeast Nadsoniella higra var. hesuelica and its physiological function. Abstract. Kida. dessert. M: 1970).

A significant part of the resistance of cells of strain 365 can confidently be associated with the existence in their cell walls, on the border with the outside world, Cherng. prophetic. (Ruban E. L., Fomin, C., Master, S. P. a Study of melanin Nadsoniella higra by the EPR method. - Reports of the Academy of Sciences of the USSR, 1968, I. 182, No. 5, S. 1219). This black pigment extracted from biomass forms and represent a basis for anticancer drug.

However, as it turned out, in addition to its main purpose "Stromelysin" strain 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 365 Antarctic black yeast Nadsoniella higra 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 intensive protivopedikuleznoe action, stopping the avalanche of available labile radicals, damaging healthy cells and tissues. The basis for this effect provide a system polyisoprene in the molecules of the additive "Stromelysin" and its inherent high soalnya of melanin, have discovered free-radical 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 radiation background 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" also helps with stomach ulcers and duodenal ulcers (ulcers fast rubouts), with XP. gastritis (no symptoms), it restores the structure of the liver and pancreas, activates the immune system "cleans" the Airways of smokers, as an adaptogen increases mental and physical servant who occupies the symptoms of XP. prostatitis with restoration of natural features and other

"Stromelysin" should be added preferably in 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. In the composition of medicinal ointments and compositions additive significantly increases or itself provokes a strong analgesic effect, as an additive to rinse quickly relieves sore throat, drink - cough etc.

The additive may be introduced into food or cosmetic and therapeutic agent as an individual substance or in combination (compositions) with other substances, including biologically active, or be introduced as a component in other supplements. Especially interesting and vitamin complexes. Thus, it was found that the Addendum is"ptx2">

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

Getting the dietary Supplement.

Strain 365 Antarctic black yeast Nadsoniella higra var. hesuelica cultivated in a known manner under conditions of aeration and agitation in a liquid nutrient medium, for example, the following composition, g/l: K2HPO 0,1; CaCl20,03; MgSO40,05; NaCl, 0,05 with glucose (2% or sucrose (1.5 to 2%), liquid yeast autolysate (1%) at 26-28oC pH 3.5-3, during the week. To increase biomass yield and stabilization of low pH in the environment, add citric acid sodium 0,5%.

In some circumstances, to grow the biomass producer strain 365 and apparatus for aeration and stirring, if cultivation is carried out in a thin layer of fluid with sufficient natural aeration, or on solid nutrient media, for example, what and where the strain grows well and easily with them is washed away. Without filters the culture grows partially on Wednesday and removed bad. Economical consumption of nutrients inherent in the cultures of strains 365, allows you to change several times filters on the same environment with additional wetting its surface.

The biomass from the liquid medium is separated by centrifugation and washed repeatedly with water, then autoclave at 0.5 ATM. 0.5 N. NaOH for 2 to 3 hours-Cell mass is removed by centrifugation, centrifugal acidified with conc. NCl to pH 2. Sediment in the form of black and brown flakes separated by centrifugation and repeatedly washed with distilled water. Dried out. So get a water-insoluble additive. To obtain water-soluble form of the additive washed precipitate is thoroughly dissolved in a small amount of 12 - 25% ammonia solution, followed by a soft evaporation of 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 koncentrirovannaya 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 can easily be dosed when added to drinks.

Examples of use of Astromania" as biologically active additives to food products, cosmetics and medicines: test-volunteers.

Example 1. Tester L, W, 1936 born., Ultrasound from 20.06.95 and 1.08.96: Liver: increased structure diffuse sealed off from the elements fatty infiltration, vascular drawing is rendered poorly. Pancreas: structure condensed considerably, coarse-grained. Additive "Stromelysin" table water "Holy spring" to 10 mg per dose 3 times a day with 15.04.97 ultrasound from 7.05.97 liver: not enlarged, the structure of the normal vascular pattern is not changed, pancreas: structure of the ordinary. At the same time stopped the constant 7-month cough is the result of XP. bronchitis as a complication of the flu and stopped aching stomach, there was no need in the daily intake of Apple cider vinegar.

Example 2. Test L. 1936 born. Herpes in the acute stage - fever, rashes on the tongue. Single Supplement 30 mg "Astronautica 4 months. Before taking supplements herpes rash appeared regularly at the slightest cooling.

The patient, 1961 born. Squamous keratinized cancer of the left lung with metastasis to lymph nodes subaortalnom and bifurcation regions T3N2Mo. 27.08.93 advanced chemistry, irradiation combined pneumonectomy left, metastasis in the area of postoperative scar, squamous cell carcinoma, excision of nodes? 7.04 baby mortality.94, the resumption of metastasis, re-excision 27.04.94, the resumption of metastasis, re-excision 27.06.94, squamous cell carcinoma, postoperative period with lymphorrhea, festering wounds, I tbsp. radiation disease variability. Written out. The connection reception "Astromania inside with water initially at 40 to 30 mg, and lotions on the wound. Accepts additive "Stromelysin" up to the present time with breaks of about 2.5 mg per day night of drinking. Last survey: February 96 g and Dec. 96, Results: without pathologies.

Example 3. He is also a persistent smoker. Bronchoscopy at the same time: no traces of smoke.

Example 4. Test N. L. J., 1938 Constantly apply the cream "Forest nymph". The addition of cream additive: leather razogrewatego joint, secondary trigeminal neuralgia II branch. Disease duration: September 1994 the Effect of obezbolevanija "Filipina" was not sufficiently severe side effects. Single Supplement "Astromania" in the water "Holy spring" in number 3, which Consists of 20 min was fully litter usually a long morning of acute pain, which was not renewed almost 52 hours When resuming pain receiving water as an additive in combination with 20 PL. "Pentavite" eliminated pain syndrome, and the effect was prolonged almost 4 days. Previously it was found that "Pentomic" and he can stop hard to bear night pain after about 1 hour, if you take 100 table. (single experiment), severe pain then returned on the 6th day. Re-admission "Pentavite in doses up to 30 PL. a day no effect on pain is not provided (attempts were made during the month).

Example 6. The old lady S. 1912 All possible Algie, rubbing in the hands of the left, especially the patient, the drug composition menthol 1 g, fenilsalitsilat 3 g vaseline oil 96 g was ineffective during acute pain; adding melanin in the composition resulted in fast - in 5 min and stokoe (10%) oil.

Thus, from comparison with known means the same destination can be seen that the strain 365 Nadsoniella nigra var. hesuelica Antarctic black yeast is the percentage of valuable highly biologically active additives in food products, cosmetics and medicines.

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

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

3. The use of natural strain 365 Antarctic black yeast Nadsoniella nigra var. hesuelica as producer melaninogenica biologically active additives under item 1 for cosmetics.

4. The use of natural strain 365 Antarctic black yeast Nadsoniella nigra var. hesuelica as producer melaninogenica biologically active additives under item 1 for medicines.

 

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