Strain of bacteria micrococcus luteus 805 - producer of site-specific methyl-dependedn endonuclease mluvi

FIELD: biotechnology.

SUBSTANCE: strain Micrococcus luteus, deposited under registration number B-12410 in The All-Russia Collection of Industrial Microorganisms of GosNIIGenetika FSUE is the producer of the site-specific methyl-dependent endonuclease MluVI. The invention provides a new MD-endonuclease enzyme which recognizes and cleaves both strands of the DNA sequence 5'-GCGC^NGCGC-3'/3'-CGCGN^CGCG-5' before the central nucleotide (N) with formation of single-nucleotide 5'-protruding ends in the presence in a given sequence of at least six C5-methylized cytosines and can be used for large-site-specific hydrolysis of DNA containing C5-methyl-cytosine bases.

EFFECT: increase the efficiency of strain application.

2 dwg, 1 tbl, 3 ex

 



 

Same patents:

FIELD: biotechnology.

SUBSTANCE: strain of bacteria Rhodococcus sp. LER-12 having the ability to dispose quickly of crude oil and petroleum products (diesel fuel, motor oil, hydraulic oil, gas condensate) is deposited in the RNCM under the registration number Rhodococcus sp. RNCM Ac-2626D and can be used for purification of soils and water from contaminations with crude oil and petroleum products, in a wide temperature range from +4 to +30°C.

EFFECT: invention enables to improve the quality of purification of soil and water from crude oil and petroleum products.

3 tbl, 5 ex

FIELD: biotechnology.

SUBSTANCE: strain is deposited in the State collection of pathogenic microorganisms "SCIM-Obolensky" under the number F-11. The culture fluid centrate obtained as a result of culturing of said the strain on the liquid nutrient medium has high activity against anthrax causative agent. The zone of inhibition of growth of Bacillus anthracis is 20-25 mm and 35-45 mm when the culture fluid centrate of the said strain is used in an amount of 10 mcl and 100 mcl, respectively.

EFFECT: strain of micromycete Trichoderma hamatum has antibacterial activity against anthrax causative agent Bacillus anthracis.

4 tbl, 3 ex

FIELD: biotechnology.

SUBSTANCE: nutrient medium comprises sodium chloride, potassium chloride, magnesium sulphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, calcium chloride, sodium bicarbonate, L-arginine, L-glutamine, L-tyrosine, L-tryptophan, calcium pantothenate, pyridoxal HCl, thiamine, meso-inositol, nicotinamide, riboflavin, folic acid, glucose, enzymatic hydrolyzate of muscle proteins, lactalbumin enzymatic hydrolyzate, sodium salt of benzylpenicillin with activity of 0.09×105-1.1×105, kanamycin sulphate with activity of 0.09×105-1.1×105, nystatin with activity of 6×104 to 6.5×104 IU/l, choline chloride, succinic acid and distilled water in a predetermined ratio of components.

EFFECT: invention enables to improve the quality of the product obtained.

1 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. The strain of fungus Aspergillus niger B-6 is the producer of citric acid. The strain Aspergillus niger is deposited in the joint Russian Collection of Agrocultural Microorganisms RAAS (RCAM) under registration number RCAM02149 and can be applied in the production of citric acid.

EFFECT: invention makes it possible to increase the output of citric acid.

2 tbl, 6 ex

FIELD: biotechnologies.

SUBSTANCE: method of production of nanodisperse additive for concrete is offered. Cyanobacteria of the sort Pseudanabaena sp. 0411 or Leptolyngbya laminosa 0412 are cultivated in a nutrient medium at the temperature 23-25°C. The nutrient medium is Z-8 medium with adding of sodium silicate solution neutralized by 2 M HCl. Ratio of solution of silicate of sodium and Z-8 5:1 medium. Cultivation is carried out in the bioreactor at continuous lighting and mixing during 10 days with the subsequent removal of nutrient medium residue. Then the culture is poured by 30% hydrogen peroxide solution and heated up to 70°C. The produced biosilicated nanotubes are washed out by distilled water with subsequent treatment in a mechanical activator in the water medium of anion surface-active substance of naphthalene formaldehyde type at the ultrasound frequency 35 kHz and concentration of solid phase 5% up to the particle size 85-250 nanometres.

EFFECT: increase of mobility of concrete mix, curing acceleration, increase of strength and density of concrete, decrease of water absorption.

3 tbl, 1 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanovermiculite and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of nitrogen-fixing and phosphate-mobilising microorganisms.

1 tbl, 14 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanobentonite and distilled water.

EFFECT: enhanced growth rate of phosphate-mobilising and nitrogen-fixing microorganisms.

1 tbl, 14 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nano sapropel and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of phosphate-mobilising and nitrogen-fixing microorganisms.

1 tbl, 15 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanozeolite and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of nitrogen-fixing and phosphate-mobilising microorganisms.

1 tbl, 15 ex

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology and concerns the strain Escherichia coli BL21(DE3)Gold/pETmin-CypA producing human recombinant cyclophilin A. The characterised strain is produced by cell transformation of the strain BL21(DE3)Gold by pETmin-CypA plasmid. The plasmid has a size of 5865 base pairs and contains XhoI-NdeI fragment of pET-22b(+) vector carrying ampR ampicillin resistance gene, a promoter and RNA-polymerase T7 phage terminator and a polylinker. A fragment produced by PCR with using the oligonucleotide primers 5'-TTATACATATGGTCAACCCGACCGTGTTCTTC and 5'-TTTCTCGAGTTATTCGAGTTGTCCACAGTCAGC of pCyPAwt/pGEX-2TK plasmid with the closed fragment of hrCpA (human recombinant cyclophilin A) gene having a size of 498 base pairs is cloned at XhoI-NdeI sites.

EFFECT: presented strain is high-producing; it requires no complicated clean-up system and can be used in cancer treatment.

2 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to molecular biology, biochemistry and genetic engineering. The invention discloses a nucleic acid characterised by a nucleotide sequence which encodes a protein consisting of an amino acid sequence with deletion, replacement or addition of one or more amino acids in the amino acid sequence SEQ ID NO: 2 or SEQ ID NO: 7 and having phosphatidic acid phosphatase activity, a corresponding protein, a recombinant vector, a protein expression cell and a method of producing a fatty acid composition.

EFFECT: invention can be used to produce polyunsaturated fatty acids in the food industry.

9 cl, 6 tbl, 7 dwg, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology. What is presented is an enzyme biocatalyst in the form of nanoscale particles for in vivo detoxification of organophosphorous compounds. The biocatalyst represents non-covalent polyelectrolyte complexes. The given complexes consist of a polyhistidine-containing polypeptide with the properties of organophosphosphorous hydrolase and polyethylene glycol-polyglutamic acid block copolymer in a charge relation of enzyme:block copolymer falling within the range of 2:1 to 1:5.

EFFECT: invention provides significant simplification of the biocatalyst technology, an increase of its catalytic efficiency, a dose decline, an immunotoxic suppression, as well as higher activity in hydrolysis reactions of pesticides and toxic materials.

4 cl, 6 ex

FIELD: biotechnologies.

SUBSTANCE: from Mortierella alpina a new gene was obtained coding phosphatidic acid phosphatase (EC 3.1.3.4), open reading frame was determined and amino acid sequence of enzyme, named as MaPAP1, was determined. Method for obtainment of recombinant protein by transformation of host cells was described, where upon use of yeast cells or cells of lipid producing fungi as host cells not only increase of total quantity of fatty acids, but change of fatty acid composition cell is provided as well.

EFFECT: increase of arachidonic acid quantity relation to total quantity of fatty acids.

8 cl, 7 tbl, 2 dwg, 8 ex

FIELD: biotechnologies.

SUBSTANCE: at fermentation of obtained strain in 3 l of a laboratory fermenter in minimal medium at the temperature of 30°C, ferment activity in culture liquid reaches the level of 210 U/ml.

EFFECT: invention allows obtaining Specific Phospholipase with high efficiency degree.

3 dwg, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely a method for preparing placental alkaline phosphatase. A method for preparing placental alkaline phosphatase consists in the fact that blood-washed human placenta is homogenised; the homogenate is extracted in a solution of potassium chloride; the extract was centrifuged, heated, centrifuged; after the extract is primarily heated, the supernatant is heated again in the presence of cadmium sulphate; the flask is quickly cooled; ammonium sulphate is added; the precipitate is separated by centrifugation; the supernatant is subjected to hydrophobic column chromatography, and a ferment eluate is cleaned by preparative polyacrylamide gel electrophoresis; the ferment gel is coloured; the alkaline phosphatase fractions are sectioned under direct vision.

EFFECT: method enables simplifying the process of preparing and cleaning the placental alkaline phosphatase enzyme with a significant yield increase.

1 tbl, 2 ex

FIELD: biotechnologies.

SUBSTANCE: peroxy hydrolase ferment is described, which may hydrolyze n-nitrophenylcaproate (pNC6) or n-nitrophenyloctanoate (pNC8) in presence of peroxide, where the specified ferment includes one of the following combinations of amino acid substitutes: Ala in the position 154 and Met in the position 194; Gly in the position 154 and Val in the position 194; or Gly in the position 12 and Met in the position 194, where the specified positions of amino acids are positionally equivalent to positions 12, 154 and 194 in the sequence SEQ ID NO: 2 of peroxy hydrolase M. smegmatis, and where the specified ferment produces peroxy acid. The following components are disclosed: produced nucleic acid, which codes the specified peroxy hydrolase ferment; recombinant nucleic acid, which expresses the peroxy hydrolase ferment, including a promotor and the specified produced nucleic acid; a vector of expression, including the specified recombinant nucleic acid; a host cell containing the recombinant nucleic acid and producing the peroxy hydrolase ferment. Compositions are described for washing, bleaching and disinfection, including effective amount of peroxy hydrolase ferment and source of hydrogen peroxide. Methods of washing, bleaching and disinfection are proposed, which include maintenance of a substrate in presence of the specified compositions for washing, bleaching or disinfection of the specified substrate, accordingly. Besides, it is suggested to use the specified produced peroxy hydrolase ferment for hydrolysis of an acyl ether substrate containing up to 8 atoms of carbon, in presence of hydrogen peroxide.

EFFECT: invention makes it possible to perform a reaction of hydrolysis of the specified compound in presence of hydrogen peroxide with formation of peroxy acids.

19 cl, 5 tbl, 4 ex

FIELD: biotechnologies.

SUBSTANCE: fusion peptide is presented for neutralisation and destruction of organophosphorous compounds, comprising a signal peptide TAT3 with amino acid sequence SEQ ID NO: 5, presented in the description, functionally connected to the sequence of organophosphate hydrolase SEQ ID NO: 18, classified as protein EC 3.1.8. The following components are described: extracted polynucleotide, which codes the specified fusion protein; a vector containing the specified polynucleotide; and a procaryotic host cell containing the specified vector and expressing the specified fusion protein. The method is proposed to produce a ferment, which destroys organophosphorous compounds, including expression of the specified polynucleotide in the procaryotic host cell and production of the specified ferment.

EFFECT: invention makes it possible to increase expression of organophosphate hydrolase in a host cell.

13 cl, 1 tbl, 9 dwg, 2 ex

FIELD: biotechnologies.

SUBSTANCE: method includes the following stages: conservation of cells in presence of buffered 80-90% glycerine, collapse of cell shells by 3% triton X-100, extraction by increasing concentrations of salts: 0.14 M, 0.35 M; 2 M NaCl, 6 M guanidine hydrochloride with 0.1% β-mercaptoethanol, extraction of positively charged proteins from above fractions with the help of ion-exchange chromatography with amberlite IRC-50 in an interrupted gradient of guanidine hydrochloride: 6%, 8.9%, 10.6%, 13% on 0.1 M potassium-phosphate buffer pH 6.8 and detection of sites of sensitivity in them to Arg-X proteolysis.

EFFECT: invention may be used in analysis of molecular-genetic mechanisms of procaryote cell structure formation and role of protein components in their organisation, and also when studying features of genome remodelling, which is necessary for opening of paths of regulation mechanisms of macro- and microorganisms.

9 dwg, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to enzymatic techniques of removing phospholipids from plant oil. The method of degumming an oil composition involves contacting the oil composition containing phospholipids simultaneously with one or more phospholipase A (PLA) enzymes in amount of about 2 ppm of the active enzyme or less and one or more phospholipase C (PLC) enzymes in amount of about 30 ppm of the active enzyme or less. Reaction of the enzymes with phospholipids takes place at pH of about 3-7, at temperature of about 40-80°C and the reaction takes less than one hour. The reaction products are separated from the oil composition. A method of degumming oil is also disclosed.

EFFECT: obtained degummed oil composition has phospholipid content, measured in parts per million of phosphorus, of about 20 ppm or less.

23 cl, 7 dwg, 7 tbl, 38 ex

FIELD: chemistry.

SUBSTANCE: disclosed are versions of mutant recombinant phytase PhyA-Cf from Citrobacter freundii bacteria: 1) a version containing an amino acid substitution D144N or D144R, or D144Q, or D144K; 2) a version containing an amino acid substitution V226A or V226G; 3) a version containing an amino acid substitution G343D or G343E, or G343K, or G343R, or G343N; 4) a version containing amino acid substitutions in positions D144 with N, R, Q or K, and in position V226 with A or G; 5) a version containing amino acid substitutions in position D144 with N, R, Q or K, and in position G343 with D, E, K, R or N; 6) a version containing amino acid substitutions in position V226 with A or G, and in position G343 with D, E, K, R or N; 7) a version containing amino acid substitutions in position D144 with N, R, Q or K, and in position V226 with A or G, and in position G343 with D, E, K, R or N. DNA fragments which code said recombinant phytase are provided. Described are strains which produce recombinant phytases Pichia pastoris PhyA-CfDVG and Pichia pastoris PhyA-CfDV, which are deposited in the Russian National Collection of Industrial Microorganisms (VKPM) under number VKPM Y-3495 and VKPM Y-3494, respectively.

EFFECT: invention enables to obtain phytases having thermally stable properties.

13 cl, 2 dwg, 10 ex

FIELD: biotechnology.

SUBSTANCE: invention is a recombinant plasmid pET40CmAP/CGL determining the synthesis of hybrid bifunctional polypeptide CmAP/CGL with the properties of highly active alkaline phosphatase of marine bacterium Cobetia marina (CmAP) and galactose specific lectin of mussels Crenomytilus grayanus (CGL). The plasmid comprises NcoI/SalI - fragment of plasmid pET-40b(+) (Novagen) and the DNA fragment of 2028 base pairs, which is a chimeric gene encoding structural genes of the alkaline phosphatase CmAP and galactose specific lectin CGL, interconnected by polynucleotide encoding the amino acid sequence of the linker (G4S)3EL and enterokinase site D4K. Also a recombinant strain of E. coli Rosetta(DE3)/pET40CmAP/CGL is provided, transformed by the indicated plasmid, - producer of the hybrid bifunctional protein CmAP/CGL, comprising the amino acid sequences of recombinant alkaline phosphatase CmAP and galactose specific lectin CGL. The method of obtaining the hybrid bifunctional protein CmAP/CGL is proposed.

EFFECT: invention enables to obtain highly purified preparation of the hybrid protein with the properties of recombinant highly active alkaline phosphatase CmAP and galactose specific lectin CGL, and can be used to detect differences in glycosylation of oncofetal antigens.

3 cl, 2 dwg, 3 ex

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