Method of restoring sensitive layer of biosensor
SUBSTANCE: invention relates to biotechnology. The method involves treating the sensitive layer of a biosensor with solution of a common fraction of protease of hepatopancreas of king crab in a buffer comprising tris-HCl 50 mM, CaCl2 3 mM, NaCl 100 mM, pH 8.0 at solution temperature of 35-40°C. The process is carried out in multiple successive steps with a solution of a solution of a common fraction of protease of hepatopancreas of king crab in said solvent at temperature of 35-40°C followed by exposure. The sensitive layer of the biosensor is successively washed with dodecyl sulphate dissolved in the same buffer in concentration of 0.1% and then with water at each step. The treatment process is carried out three times.
EFFECT: invention reduces the duration of the process.
The invention relates to the field of biosensor analysis and relates to a method of recovery of the sensitive layer of the biosensor.
The biosensor chips are a high-tech product, which leads to the fact that non-factory of their production is impossible. At the same time they are essentially disposable items, reuse, which is not envisaged, although the cost of the chip is high enough, and the cost of one analysis can be very significant. In many cases this is not an obstacle for the use of biosensors as when performing a vital analysis or the development of new drugs price analysis is not the determining factor. At the same time there are studies, for which you need to spend tens or even hundreds of chips. In many cases this is not possible for economic reasons.
In most cases, if immobilized component in the study of the interaction is a protein or polypeptide, its immobilization on the chip is made by the user. From the factory chips are produced without immobilizovannoi components, but only prepared for immobilization. This is because protein and polypeptides immobilized on the chip surface stored worse than separately chips and sewn components, for which the nutrient is possible to provide optimal storage conditions for each.
Immobilizovannyi component must be put firmly on the surface of the chip. Otherwise, if this component is washed away from the chip, there is a strong drift, interfering with the measurements. In addition, the loss of the immobilized component leads to a continuous decrease in the sensitivity of the biosensor. Therefore, usually used for immobilization irreversible presume, leading to the formation of covalent bonds between the substrate of the chip and immobilizovannoi molecule. Most often used immobilizatsiya for the amino group on carboxymethylamino dextranase substrate activated with a mixture of EDC/NHS (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide -hydroxysuccinimide). In addition, it is possible to commit macromolecules for carboxyl group, sulfhydryl group, etc. Using any of these methods leads to irreversible fixation of macromolecules, after which they remain on the surface of the chip and cannot be washed off with her, detergents, solvents and other cleaning agents. Therefore, over time, after degradation of the immobilized component used chip already unfit for any further use. At the same time, the substrate of such a chip is quite suitable for the immobilization of other macromolecules, but to use it to re-immobilization in most cases, it is not because some places (usually Carbo is strong groups) immobilization deactivated or occupied in the previous landing, and the remaining from the previous immobilization of molecules can lead to unpredictable artifacts and errors of measurement.
To get used chip in a state close to that in which it comes from the factory, it is necessary to more fully resolve with its surface remains of previously immobilized macromolecules and, if necessary, to restore to the original values of the surface density of carboxyl groups. At the moment there is no way aiming to break the covalent bond formed during immobilization, however, have the ability to delete all items sewn macromolecule in addition, which is directly sewn to the substrate.
This can lead to the surface of the chip in contact with a solution of enzyme able to hydrolyze communication between the components of macromolecules. For proteins, and proteins should be used protease, and nucleic acids - nuclease - Rncse, Tnkz[1, 2, 5].
There is a wide choice of proteolytic enzymes, but not all of them are suitable for carrying out regeneration , as most proteases cut the peptide bond only on certain amino acids, which, in addition, must be available for exposure of the enzyme, that is located outside of the protein globule. Such proteases are not able gidrolizovat is sewn proteins. Always remain areas that do not contain the right amino acids, and which will not be hydrolyzed and removed. In addition, even if the protein molecule will be cut in several places, the cut pieces will not be separated from the protein, because the protein chain in globule strongly intertwined with itself, and cut off pieces can't leave globule.
There are enzymes that can cut from the protein chain amino acid for amino acid with a C or N end (this is different enzymes, respectively-peptidases and N-peptidases), which in principle can hydrolyze the entire protein residue. However, such enzymes are still not solve the problem for several reasons. There are proteins in which the C-end, N-end, or both hidden inside the protein .
Such proteins are invulnerable to the appropriate type of end proteases or to both. Because in most cases, the immobilization of a protein globule is not at the end of the chain, and for any internal link for complete cleaning sensitive surfaces of the biosensor must be present in regenerating the solution of both types of end proteases. At the same time, these proteases are normally resistant to its effects, but vulnerable to proteases of another type. And finally, it is quite expensive proteins which are not always available for purchase. At the same time, given the other activities of such enzymes is generally low, and for effective removal of proteins within an acceptable time they need to apply in large numbers.
There is a method of regeneration of optical elements of the biosensor by hydrolysis of the immobilized protein with a solution of proteolytic enzyme Pronase that is, the Solution of the specified enzyme concentration 1 mg/ml were prepared in buffer solution composition is 20 mm phosphate, 150 mm sodium chloride, pH 7,2.
The chip was placed in the solution for about 15-17 hours at room temperature. Then it was rinsed in distilled water. There was obtained a satisfactory level of recovery properties of the chip, however, the main drawback of the method is the high consumption of proteolytic enzyme, its high cost (http://www.sigmaaldrich.com/life-science/biochemicals/biochemical-products.html?TablePage=16410626 and the lack of production of this drug. In this regard, the cost recovery of the optical chip by this method is too high .
In accordance with the invention describes a method of restoring the sensitive layer of the biosensor by multistage processing solution the total fraction of proteases hepatopancreas crab in a buffer solution composition of Tris-HCl 50 mm, CaCl23 mm, NaCl 100 mm, PH 8.0 at a solution temperature of 35-40° C, followed by exposure and sequence is through washing sensitive layer first dodecyl sulfate and then with water after each stage.
The total fraction of proteases from hepatopancreas crab is a very effective set of enzymes designed and are highly adapted for destruction and transformation into oligopeptides and single amino acids in any protein molecules. This mixture is able to hydrolyze any proteins regardless of their primary and secondary structure, with a very high speed and at a temperature from +2°C and above. These properties are determined by habitat conditions and physiology of the red king crab and optimized over millions of years of evolution. The total fraction of proteases is a by-product in the processing of red king crab in the food products, so the cost is not high .
According to literature data , the optimal conditions for storing and actions enzyme preparation collagenase crab are the following: for storage - potassium phosphate 100 mm, 50 mm potassium citrate (pH of 4.8); and to work Tris-HCl 50 mm, CaCl23 mm, NaCl 100 mm, (pH 8.0). The best activity of the drug is achieved at a temperature of 35-40°C.
The objective of the invention is to develop a method to restore sensitive layer of the biosensor.
Directly the process of regeneration of the sensitive layer of the biosensor is as follows.
Used the biosensor chip is released from the protective shell, if it exists. Sachemship placed in a hermetically closed container minimum volume, enough to chip fit in it entirely. Prepare a solution of the total fraction of proteases of hepatopancreas crab in a buffer solution composition of Tris-HCl 50 mm, CaCl23 mm, NaCl 100 mm (pH 8.0). The temperature of the solution is maintained within the range of 35-40°C. For this purpose a portion of the dry lyophilized drug is dissolved to a concentration of 0.1-0.01 mg/ml Smaller values result in longer procedures, and large waste of the drug. The resulting solution is poured into the container placed in a chip so that the sensitive surface of the chip was coated with the resulting solution, and the lid of the container tightly closed. The container is placed in a thermostat at a temperature of 35-40°C. After 24 hours exposure, the chip is removed from the container with a solution of protease and washed in a solution of sodium dodecyl sulfate for one hour, then 5 minutes in running distilled water and again placed in a solution of protease. This cycle is repeated three times, after which the chip was washed for one hour in flowing distilled water. Thus, the total duration of the recovery of the chip is about 76 hours. The solution proteases hepatopancreas crab saves suitability to work within one month from the date of dissolution in case of storage at a temperature not exceeding 8°C [6, 7].
After okonchatelnaya optical surface of the chip blow clean compressed air to remove dust, return the chip in a protective shell and placed in a plastic bag for storage. To assess the quality of recovery chip is placed into the sensor and make sure that the provisions of the lines sensogram correspond to the position of the lines characteristic of intact dextran or approaching such. To estimate the number of carboxyl groups they control preconcentration any reference lipid. If the speed of preconcentration lipid considerably reduced, it is possible to restore the number of carboxyl groups the following procedure:
The whole procedure of regeneration sensitive surfaces can be produced directly in the biosensor, as the applied solutions (solution proteases, a solution of dodecyl sulfate and water) do not cause corrosion and do not destroy plastics, or held separately in the supporting tubes or specially adapted containers. When conducting regeneration outside of the device removed restrictions on the duration of operations and requirements decrease of the specific activity of the protease complex. The duration of the procedures you can bring up to several days without compromising performance.
Experimental verification showed that the total fraction of proteases hepatopancreas crab quickly and effectively removes sensitive surface of the biosensor chip more than the 99% of the previously immobilized protein. Thus liberated from the results of the previous planting chip it is possible to plant a new batch of protein not less than 90%, which sits on a new chipmoondang increase the number of carboxyl groups on the dextran chip is not needed.
Used in the described method, the total fraction of proteases hepatopancreas crab has high proteolytic activity and low cost, because it is produced in large quantities as a by-product of the food industry.
1. Patent RU No. 2027181.
2. maxXbond: the First regeneration system for twisted DNA-columns https://www.applichem.com/ru/o-produktakh/maxxbond-pervaja-sistema-regeneracii-dnk-kolonok/.
3. Handbook of Proteolytic Enzymes / Eds Barrett AJ, Rawlings N.D., Woessner J.F. London: Acad. Press, 1998.
4. Ronald C. Chatelier, Thomas R. Gengenbach, Hans J. Griesser, Michael Brigham-Burke, t and Daniel J. O Shannessyt'(1995). A General Method to Recondition and Reuse BIAcore Sensor Chips Fouled with Covalently Immobilized Protein/Peptide. ANALYTICAL BIOCHEMISTRY 229, 112-118.
5. Patent RU No. 2239827.
6. THE 2639-001-45554109-98. The drug, the enzyme collagenase from hydrobionts.
7. Patent RU No. 2280076.
1. The method of restoring the sensitive layer of the biosensor by treating it with a solution of enzyme, characterized in that the quality of an enzyme solution used solution the total fraction of proteases hepatopancreas red king crab in the buffer composition of Tris-HCl 50 mm , CaCl23 mm, NaCl 100 mm, pH 8.0 at a temperature of the solution is in the range of 35-40 °With, and the process is carried out in several successive stages by processing the sensitive layer of the biosensor solution the total fraction of proteases hepatopancreas red king crab in the specified solvent at a temperature in the range of 35-40°With subsequent exposure and sequential leaching sensitive layer dodecyl sulfate dissolved in the same buffer at a concentration of 0.1 %, then the water at each stage.
2. The method according to claim 1, characterized in that the loop processing is carried out three times.
SUBSTANCE: invention relates to biotechnology. Claimed is a method of obtaining a liquefied biomass, including obtaining the biomass material from sugar beet and/or sugar cane, liquefying the said biomass with enzyme mixture. The said enzyme mixture is used in an amount of at least 0.025% of the biomass. The enzyme mixture includes cellobiohydrolase, beta-glucosidase and polygalacturonase and additionally one or several enzymes with hemicellulase activity, selected from arabinase, xylanase, pectinmethylesterase, ramnogalacturonase and 1,3-/1,6-beta-D-glucanase. Resulting product contains less than 2 wt % of a residual insoluble dry substance.
EFFECT: liquefied biomass is stable in storage and can be used for obtaining products resulting from fermentation, such as ethanol, butanol, acetone, 1,3-propanediol, propanol, acetic acid, lactic acid and propionic acid.
14 cl, 7 dwg, 3 ex
SUBSTANCE: invention relates to a method of identifying improved protein versions. The method includes testing a multitude of protein versions with single substitutions in the first test of the first property and in the second test of the second property. The parent protein property is given a value 1.0 in each test. The favourable first or second property possesses a value, exceeding 1.0, and extremely unfavourable first or second property has a value less than approximately 0.80. The said parent protein represents protease. The first property is stability, the second property is washing efficiency. Identification of substitution and introduction of the substitution into the protein to obtain the protein with multiple substitutions are carried out. The substitution does not interact in a three-dimensional structure of the said parent protein, and one of the substitutions contains a change of the total charge, equal to 0, -1 or -2 with respect to the parent protein, and, at least, one of the substitutions contains the change of the total charge, equal to +1 or +2 with respect to the parent protein. The protein version with multiple changes in the first and second tests is tested and the said protein version is identified.
EFFECT: invention makes it possible to obtain the protein version with improved stability and washing efficiency.
14 cl, 6 dwg, 6 tbl, 7 ex
SUBSTANCE: invention relates to field of biochemistry and biotechnology. Performed are: crushing and sieving of lignocelluloses material and selection of granules with particle size from 0.08-0.1 mm. As lignocelluloses material used are: straw, grass, chip, corncobs, bagasse and oil cake. Obtained granules are mixed with weight ratio granules-water 1:(1-5) at temperature 70-90°C. They are dispersed through colloid mill for 1-2 hours to obtain suspension with particle size 40-80 mcm. Homogenisation of obtained suspension is performed under pressure 50-100 atm and temperature 60-85°C for 1-2 hours. Suspension with particles with size 10-40 mcm is obtained. Bufferisation of obtained suspension with buffer solution of sodium acetate and acetic acid with pH value = 4.8-5.8 is carried out. Mixture of enzymes: cellulose in amount 10-60 international units per gram of lignocelluloses material, β-glucosidase in amount 40-100 international units per gram of lignocelluloses material and xylanase in amount 60-120 international units per gram of lignocelluloses material is added. Mixture of enzymes is introduced into reactor after cooling suspension to the temperature of carrying out enzymolysis 40-55°C. Enzymolysis is carried out in reactor for 36-72 hours at rate of reactor rotation 80-160 revolutions per minute. Content of sugar in hydrolysate is determined by method of liquid chromatography.
EFFECT: invention makes it possible to carry out processing of lignocelluloses material at low temperature 40-55°C.
7 cl, 3 dwg, 3 ex
SUBSTANCE: at first, gel filtration of the solution of Central Asian cobra venom (Naja naja oxiana) is carried out on superfine Sephadex G-75, followed by chromatography of endonuclease on SP-Sephadex C-25, dialysis of the isolated enzyme with the addition of ballast protein, sterilisation and lyophilisation. At that prior to dialysis BSA is added to the final concentration of 1.0-2.0 mg/ml, and the dialysed endonuclease without concentration is sterilised and lyophilised.
EFFECT: method enables to increase the yield of endonuclease while isolation, as well as to eliminate the obstacle to its use for treatment of cow rabies.
SUBSTANCE: invention relates to biotechnology. The method of producing globotriose involves enzymatic transfer of a galactose monomer from a galactose donor to lactose acting as an acceptor. The galactose monomer donor used is β-galactosyl azide. The enzyme used is α-1,4-galactosyl synthase, which is obtained by introducing two substitutes D327G and P402D in corresponding positions of the amino acid sequence of the wild-type alpha-galactosidase gene isolated from Thermotoga maritima. The reaction is carried out at 37°C and pH 5.0 until complete splitting of β-galactosyl azide.
EFFECT: invention increases output and purity of globotriose.
2 cl, 2 dwg, 1 ex
SUBSTANCE: invention refers to medicine and veterinary science. A method provides using bacterial 7P ribonuclease for increasing the effectiveness of suppression of RNA- and DNA-containing virus replication for treating acute viral diseases in mammals and warm-blooded animals.
EFFECT: Bacillus intermedius RNAase suppression of virus replication is ensured by using substantially lower doses of enzymes introduced into an infected body with the increased effectiveness of virus replication and therapeutic process.
2 cl, 8 tbl, 7 ex
SUBSTANCE: invention refers to yeast strain Yarrowia lipolytica All-Russian collection of industrial microorganisms Y - 3852 - producer of phytase. Recombinant strain is obtained by transformation of strain Yarrowia lipolytica POlf ATCC MYA-2613 and includes phytase gene PhyA Obesumbacterium proteus.
EFFECT: strain can be used in animal breeding for obtaining feed additives.
2 dwg, 5 ex
SUBSTANCE: recombinant bacteria strain Escherichia coli All-Russian collection of industrial microorganisms B-11271 - producer of hydrolase of esters of alpha-amino acids from Xanthomonas rubrilineans All-Russian collection of industrial microorganisms B-9915 is built, which is obtained by transformation of strain-recipient Escherichia coli BL21(DE3) of plasmid DNA corresponding to nucleotide sequence SEQ ID NO 1 containing gene aehR of hydrolase of alpha-amino acids esters from Xanthomonas rubrilineans All-Russian collection of industrial microorganisms B-9915, which is not modified on 5'-end with the sequence coding a polyhistidin tail and being under control of promoter T7, as well as resistance gene to kanamycin Kan.
EFFECT: invention allows obtaining hydrolase of esters of alpha-amino acids with high efficiency degree.
2 cl, 3 dwg, 1 tbl, 7 ex
SUBSTANCE: skins of pond fish are flushed with cold flushing water during 10-15 minutes. They are crushed to the size of 2-3 mm. Water extraction is performed at the temperature of 40-45°C during 40-50 minutes at the ratio of crushed skins to water, which is equal to 1:1 at periodic mixing. Then, they are filtered; liquid fraction is dried in a spraying drier at the drier outlet product temperature of 60-65°C during 15-25 minutes so that hyaluronic acid is obtained. Solid fraction is subject to bleaching during 12 hours with hydrogen peroxide-salt solution that is prepared by mixing of 1 l of 3% hydrogen peroxide and 20 g of sodium chloride. Treatment of bleached solid fraction is performed with 1.0-1.2% solution of sodium hydroxide during 24 hours at the temperature of 20-25°C with further neutralisation of the obtained mixture with 3% boric acid solution. Treatment of swollen solid fraction is performed with Pancreatin ferment preparation solution taken in the quantity of 0.5-0.6% to the weight of solid fraction during 1.5-2.0 hours at the temperature of 37-40°C. Flushing of solid fraction is performed with cold flushing water for removal of Pancreatin residues so that collagen is obtained. The obtained collagen, depending on the purpose, is supplied for drying in drying chambers with forced air circulation at the temperature of 18-20°C during 12 hours and storage in dry ventilated rooms at the temperature of not higher than 20°C during 24 months or frozen to the temperature of minus 18 - minus 20°C and stored at the temperature of minus 18 - minus 20°C during 24 months. The liquid fraction dried in the spraying chamber is stored at the temperature of 0-4°C during 12 months or dissolved in physiological buffer solution.
EFFECT: improvement of the method.
2 dwg, 1 tbl, 1 ex
SUBSTANCE: strain is built by transformation of strain-recipient Escherichia coli BL21 (DE3) with plasmid DNA containing the coding area of gene aehR of hydrolase of alpha-amino acids esters Xanthomonas rubrilineans All-Russian collection of industrial microorganisms B-9915 under control of promoter T7 and gene of stability to kanamycin Kan of hydrolase of alpha amino acids esters from Xanthomonas rubrilineans All-Russian collection of industrial microorganisms B-9915. The proposed synthesis method of hydrolase of alpha-amino acids esters is implemented by cultivation of strain-producer Escherichia coli All-Russian collection of industrial microorganisms B-11246 with addition to the composition of environment of kanamycin and isopropyl-"в"-D-tiogallactoside as an expression inducer of the obtained product.
EFFECT: high biosynthesis level of target product, which can be used for development of methods for obtaining biocatalysts of synthesis processes of aminopenicillins and aminocephalosporins.
2 cl, 3 dwg, 1 tbl, 5 ex
SUBSTANCE: method of obtaining an antiviral preparation is carried out by preparation of an inoculation mycelium of basidiomycete Enokitake Flammulina velutipes (Curtis) Singer, preparation of a liquid nutritional medium, which contains water, vegetable oil and molasses as a carbon source, as a nitrogen source - corn flour, as mineral salts - potassium dihydrophosphate and magnesium sulphate, its sterilisation, inoculation of the sterile nutritional medium with the prepared inoculation mycelium, cultivation of basidiomycete in it in an aerobic conditions; the obtained submerged culture is divided into basidiomycete biomass and a culture liquid, from which a clot is separated by addition to the latter of ethyl alcohol, which is pressed, dried and crushed with obtaining the antiviral preparation under specified conditions.
EFFECT: method makes it possible to simplify technological process, increase the output of the antiviral preparation, possessing higher activity.
2 cl, 1 tbl, 4 ex
SUBSTANCE: strain of fungus Aspergillus oryzae Amy T-52-3-21 produces maltogenic α-amylase, and it is deposited in the All-Russian Collection of Microorganisms Institute of Biochemistry and Physiology of Microorganisms n.a. GK Scriabin RAS under the number F-4476D. The strain is made on the basis of strain Aspergillus oryzae of All-Russian Collection of Microorganisms F-3927D using the genetic engineering methods. Activity of α-amylase at 120 h of growth of the strain is 600-640 units/ml.
EFFECT: higher level of activity of maltogenic α-amylase.
1 tbl, 2 ex
SUBSTANCE: invention relates to biotechnology. Claimed is strain of Fusarium sambucinum, deposited in VKPM collection under number F-1161. Claimed strain is producent of protein food biomass.
EFFECT: invention makes it possible to accumulate biomass with high protein content with higher quantity of valuable unsaturated fatty acids, complete in composition.
2 tbl, 3 ex
SUBSTANCE: invention relates to biotechnology. Claimed is method of obtaining food fungal biomass with high protein content. Multicycle deep cultivation of Fusarium sambucinum All-Russian collection of industrial microorganisms F-1161 on liquid nutritional medium, containing sources of carbon, nitrogen, mineral salts, separation and drying of wet fungus biomass are carried out. Cultivation is performed at pH from 3.5 to 7.0 under conditions of air aeration from 0.5 to 2.0 l/l/min. Temperature mode in each cycle of fermentation is supported from the beginning of the cycle to the point of switch at the level from 26 to 30°C, and further to the end of the cycle at the level from 22 to 25°C. Point of switch is determined by accumulation of biomass to concentration from 45 to 60% from maximally achievable in fermentation apparatus, or point of switch is determined by concentration of dissolved oxygen by its reduction to the value from 20 to 40% of saturation ( calculated per atmospheric air pressure).
EFFECT: invention makes it possible to obtain the largest accumulation of biomass with high protein content with specified quantity of nucleic acids.
SUBSTANCE: mutant strain is obtained by impact on Glarea lozoyensis ATCC 20957 strain by nitrosoguanidine and it is deposited in CGMCC with number CGMCC 2933.
EFFECT: fungi strain has stable genetic and productive properties, produces low quantity of impurities during fermentation and is acceptable for commercial production of antibiotic.
6 cl, 2 dwg, 2 tbl, 3 ex
SUBSTANCE: detection method of microfungi Coccidioides posadasii 36 S and Coccidioides immitis C-5 in vitro involves pre-growth of culture in mycelial phase, preparation of a suspension corresponding to 5 units of activity of a standard opacity sample, possibility of spherules formation and detection of spherules filled with endospores. Culture in mycelial phase is grown during 3 days. Possibility of spherules formation is provided by infection of the one-day culture of cells of murine splenocytes, which is obtained on RPMI-1640, and further cultivation during 5 days at the temperature of 37°C, with content of CO2 in atmosphere of 5%. In order to detect spherules in the form of round double-outline formations filled with endospores, a sample is taken, deactivated with formalin and investigated by means of a light microscopy method.
EFFECT: invention allows simplifying the method and reducing the investigation period.
SUBSTANCE: production method of keratinase provides for directed adaptation of strain Penicillium citrinum PC-54-91"ВИЛАР" by three-time subcultivation on Czapek agar medium containing 2% of hair keratin as a carbon source. Cultivation under deep conditions on Czapek medium containing 10% of hair keratin and 0.5% of saccharose during 6 days. Separation of biomass from culture fluid with further extraction of target product by two-stage chromatographic cleaning involving gel filtration on TSK-Gel TOYOPEARL HW-40 and affine chromatography on protein A to sepharose CL-4B with further lyophilisation.
EFFECT: invention allows increasing ferment yield, obtaining keratinase preparation decomposing the hair α-keratin, and improving cleaning degree of ferment preparation.
2 tbl, 5 ex
SUBSTANCE: proposed method provides for preparation of inoculating mycelium of basidiomycete, which has been chosen from the following groups: Flammulina velutipes (Curtis) Singer and/or Hericium erinaceus (Bull.) Pers. Preparation of culture medium containing pulverised cold-pressed sunflower cake, soya flour, potassium dihydrogen phosphate, magnesium sulphate and water in the specified ratios. Inoculation of the obtained culture medium with the inoculating mycelium of basidiomycete, cultivation of basidiomycete with further separation of mycelium biomass and extraction of an antitumour agent from the biomass.
EFFECT: invention allows improving an environmental situation due to utilisation of production wastes of food industry.
3 cl, 1 tbl, 4 ex
SUBSTANCE: strain Aspergillus oryzae RCAM01135 is a producer of proteolytic and amylolytic ferments, which has ability to produce proteolytic and amylolytic ferments. It was deposited at GNU VNIISKHM with the following registration number: RCAM01135. It can be used at production of different food products (fermented spices, additives, and drinks).
EFFECT: invention allows increasing the growth speed and intensity of spore formation.
4 tbl, 2 ex
SUBSTANCE: strain of unicellular algae Dunaliella salina IPPAS D-295-producer of biologically active substances having antioxidant activity was deposited in the culture collection of microalgae of the Institute of Plant Physiology n.a. K.A.Timiryazev RAS (IPP RAS (IPPAS)) under registration number IPPAS D-295 and can be used to produce biologically active substances having antioxidant activity and antagonistic activity against opportunistic pathogenic bacteria.
EFFECT: invention enables to increases the yield of antioxidant substances.
3 tbl, 2 ex
FIELD: biotechnology, in particular reagent for structural protein hydrolysis.
SUBSTANCE: method for production of protheolytic reagent includes cultivation of producer strain selected from dermatophyte of species Trichophyton mentagrophytes, Trichophyton verrucosum, Trichophyton equinum, Microsporum canis, Microsporum gypseum, preferably from fungus strains capable to produce complex of structural protein proteinases (scleroproteases) with total protheolytic activity at least 0.7 U/mg including collagenase 0.1-4.5 U/mg; keratinase 0.1-0.5- U/mg; elastase 0.5-1.9 U/mg. Cultivation is carried out, for example, in wort agar-agar or in wort broth for 20-24 days.
EFFECT: scleroproteases with improved specific activity; method for protheolytic cleavage of specific substrates (scleroproteins) with increased rate and depth.
2 dwg, 12 ex