Method of obtaining fungal protein biomass. RU patent 2511041.
 Method of accelerated growth of staphylococcus aureus for diagnostics of infections associated with delivery of health care / 2511031Growing of Staphylococcus aureus is carried out in a nutrient medium containing yolk-salt agar. At a stage of preparation for analysis the growth stimulators of Staphylococcus aureus are introduced into the nutrient medium in the form of aqueous solutions at concentrations of 10-4-10-6 wt %. The following compounds are used as growth stimulators: tris(2-hydroxyethyl)ammonium 4-chlorophenyl-sulfanylacetate or tris(2-hydroxyethyl)ammonium 2-chlorophenyloxyacetate or tris(2-hydroxyethyl)ammonium 2-methyl-4-chlorophenyloxyacetate or tris(2-hydroxyethyl)ammonium 1-benzylindol-3-yl-sulfanylacetate. |
 Recombinant strain of escherichia coli tg1(prvmoscow3253g-l) for obtaining set of pcr-standards and set of pcr-standards for determination of concentration of rabies virus "moskva 3253" in rabies antigen / 2511029Invention relates to biotechnology and deals with recombinant strain of E. coli TG1(pRVMoscow3253G-L) for obtaining PCR-standards for quantitative determination of cDNA of rabies virus of strain "Moskva 3253".Recombinant strain is created on the basis of strain of E. coli TG1 by transformation with plasmid pRVMoscow3253G-L. Plasmid is obtained by ligation of fragment G-L of the region of genome of fixed rabies virus of strain "Moskva 3253", which has sequence SEQ ID NO1, into plasmid pGem-T. Also claimed is set of PCR-standards for quantitative determination of cDNA of rabies virus of strain "Moskva 3253" in rabies antigen. Set contains solutions of plasmid pRVMoscow3253G-L DNA in concentrations 108, 107, 105, 103 GE/ml. Concentration is determined by method of polymerase chain reaction, with hybridisation-fluorescence account of results. |
 Method of determining hexoses in supramolecular structures of cells in escherichia coli / 2510846Invention relates to biochemistry and molecular biology. Conservation of cells of Escherichia coli in presence of buffered 80-90% glycerol is performed. Cell envelopes are removed with 3% triton X-100. Cell supramolecular structures are successively extracted with increasing concentrations of salts: 0.14 M (bacterioplasm), 0.35 M (loosely linked with cell residue), 2 M NaCl (strongly linked with cell residue), and 6 M guanidine hydrochloride with 0.1% β-mercaptoethanol (cell residue). Acid hydrolysis is carried out in said fractions. Anthrone method is carried out, with preliminary purification of anthrone preparation. Calibration graph is built and quantity of hexoses is determined by means of calculation formula. |
| Nutrient medium for growing legionella / 2510828 Invention is production of the nutrient medium, which creates optimal conditions for growing legionella, comprising: enzymatic hydrolyzate of pig lung, enzymatic hydrolyzate of chicken egg yolk, potassium monophosphate, trihydrate disubstituted potassium phosphate, L-cysteine hydrochloride, activated carbon, microbiological agar and distilled water at a predetermined ratio of ingredients. |
| Nutritive medium for extraction of lactobacteria / 2510416 Nutritive medium includes lactoserum, yeast autolysate, acetic acid 70%, agar and cabbage brew at the specified component ratio. |
 Method for biosorption purification of water from heavy metal ions using saccharomyces cerevisiae yeast / 2509734Invention relates to biotechnology and can be used in biological treatment of waste water from electroplating plants from heavy metal salts. The method involves adding yeast biomass to waste water, said biomass being in form of brewery wastes containing a combination of yeasts of different strains of Saccharomyces cerevisiae with viability of 90-95% in a given amount. The biomass is mixed with the waste water to obtain a suspension. The obtained suspension is held for 8 hours at temperature of 10°C-29°C and solution pH of 5.5-8.0, followed by recycling spent yeast containing heavy metals by treating with lime Ca(OH)2, with the ratio of yeast biomass to lime of 1:5-8, to obtain a mixture. The obtained mixture is subjected to wet treatment at temperature of 90°C for 1 hour, followed by isolation of the obtained mixture, which contains heavy metals, in concrete paste. |
 Diagnostics method of sensitivity of mycobacterium tuberculosis strains to injection antituberculous preparations of reserve row (aminoglycosides and capreomycin) / 2509158Invention presents a diagnostics method of sensitivity of M. tuberculosis (MBT) to injection antituberculous preparations of a reserve row. The method involves a DNA extraction stage, amplification of the investigated DNA sections by means of a polymerase chain reaction method and analysis of conformational polymorphism of single-chain fragments (SSCP). Gene section rrs is amplified in 50 mcl of a reaction mixture with addition of 5 mcl of the specimen. Promoter gene section eis is amplified in 30 mcl of the reaction mixture containing direct 5'CGGAGCCGTCGGGGTATGC and reverse 5'GCCGCGGCCAGTAGGAACA primers and 3 mcl of the specimen as per the amplification programme: 1-st stage - 95°- 4 min; 2-nd stage - 95° - 20 sec, 59° - 30 sec, 72° - 20 sec (30 cycles); 3-rd stage: 72° - 4 min; 10° - storage. Separation of amplification products in the ratio of 4 mcl of the specimen and 6 mcl of a denaturing dye is performed by electrophoresis in 8% polyacrylamide gel with 5% glycerin at voltage of 400 Volts during 5 hours at 8°C. Gel painting is performed by means of caustic silver. |
 Association of strains of bacteria-oil decomposers, and remediation method of oil-contaminated objects / 2509150Invention proposes an association of strains of bacteria-oil decomposers, which have been extracted from oil-contaminated soil, Acinetobacter species B-1037, Pseudomonas species B-989, Bacillus species B-1040, deposited at The State Research Centre of Virology and Biotechnology VECTOR. Besides, at least 30% bacteria of each strain is contained in the association. Remediation of oil-contaminated soils includes water suspension of lyophilic dried biomass of the strain association based on 109 cells per square metre. Strains of the association can utilise a wide range of oil components at the temperature of 10-15°C. |
| Bacillus subtilis subsp subtilis strain having apparent antagonism in relation to escherichia coli, salmonella typhi, staphylococcus aureus, listeria monocytogenes and resistance to streptomycin and tetracycline / 2509149 Bacillus subtilis subsp.subtilis BKM B-2711D strain has apparent antagonism in relation to Escherichia coli, Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes, and resistance to streptomycin and tetracycline antibiotics. It is deposited in the All-Russia Collection of Microorganisms of the Institute of Biochemistry and Physiology of Microorganisms Named after G.K. Skryabin of the Russian Academy of Science (IBFM RAN) and has the following registration number: BKM B-2711D, and can be used at production of probiotic bacterial preparations that can be used in veterinary medicine. |
| Bacillus amyloliquefaciens bkm b-2714d strain having apparent antagonism in relation to salmonella typhi, staphylococcus aureus, listeria monocytogenes and resistance to tetracycline and trimethoprim / 2509148 Bacillus amyloliquefaciens BKM B-2714D strain has apparent antagonism in relation to Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes, and resistance to tetracycline and trimethoprim antibiotics. It is deposited in the All-Russia Collection of Microorganisms of the Institute of Biochemistry and Physiology of Microorganisms Named after G.K. Skryabin of the Russian Academy of Science (IBFM RAN) and has the following registration number: BKM B-2714D, and can be used at production of probiotic bacterial preparations that can be used in veterinary medicine. |
 Producing active highly phosphorylated human lysosomal sulphatase enzymes and use thereof / 2510820Invention relates to biotechnology. Disclosed is a purified preparation of recombinant human N-acetylgalactosamine-6-sulfatase (GALNS) enzyme, where said enzyme includes an amino acid sequence which is at least 95% identical to amino acids 27-522 SEQ ID NO:4, which is suitable for treating a subject suffering from a lysosomal storage disease associated with GALNS, where: (a) said GALNS enzyme preparation has purity of at least about 95% as determined by Coomassie Blue staining when subjected to SDS-PAGE under non-reducing conditions; and (b) the cysteine residue at position 79 of at least 50% of molecules of the GALNS enzyme in said GALNS enzyme preparation is converted to Cα-formylglycine (FGly); where said GALNS enzyme is N-linked glycosylated at the asparagine residues at positions 204 and 423, wherein at least about 50% of the oligomannose chains attached to the asparagine residue at position 204 are bis-phosphorylated. Disclosed is a method of treating a subject suffering from mucopolysaccharidosis type IVa (MPS IVa), Morquio A syndrome or multiple sulfatase deficiency (MSD), which involves administering a therapeutically effective amount of said purified preparation of recombinant human GALNS to the subject. |
 Glarea lozoyensis mutant strain and its application / 2507252Mutant strain is obtained by impact on Glarea lozoyensis ATCC 20957 strain by nitrosoguanidine and it is deposited in CGMCC with number CGMCC 2933. |
 Antibodies against tenb2 constructed with cysteine, and conjugates, antibody-drug / 2505544There proposed is an antibody specific to TENB2 containing light and heavy chains. Heavy chain contains substitution for free (reaction capable) cysteine A121C that corresponds to A114C (Kabat numbering) or A118C (Eu numbering). Conjugate versions are proposed for prostate cancer treatment containing antibody covalently bound to auristatin, also by means of linker. The following is described: pharmaceutical composition for prostate cancer treatment that uses as active beginning the antibody or its conjugate; product for prostate cancer treatment on the basis of such composition. The invention proposes: method for defining protein TENB2 in the sample - on the basis of antibody as well as analysis for revealing prostate cancer cells at mammal and method for cell proliferation inhibiting on the base of antibody conjugate and auristatin. There described is the method for obtaining antibody conjugate (Ab) and auristatin (D) with expression Ab-(L-D)p, where p is equal from 1 to 4, and L is linker. |
 Improved expression vectors of mammals and their use / 2502800Expression vector includes: (a) replication origin OriP obtained from Epstein-Barr virus (EBV), where replication origin contains: 1) symmetry element of the second order (DS); and 2) duplication section (FR) that contains fixation point EBNA; (b) replication origin SV40; (c) insertion section for inserting a gene of concern; (d) promoter EF-1b functionally bound to the insertion section; (e) poly-A signal; (f) bacterial replication origin; (g) selected marker; and unnecessarily containing (h) sequence of nucleic acid, which codes constant area of heavy or light chain of antibody, which is functionally bound to the insertion section. With that, replication origin OriP is bound to an initiation factor of replication EBNA 1, which acts from outside and is not coded with an expression vector. |
 Method for obtaining recombinant core protein of hepatitis e virus and recombinant vaccine for prophylaxis of hepatitis e virus / 2501809Invention proposes a method for obtaining recombinant core protein of hepatitis E virus (rtHEV-ORF2) and recombinant vaccine for prophylaxis of hepatitis E virus. Core protein is obtained by cultivation of recombinant yeast strain Hansenula polymorpha "КБТ"-11/pHEV-001, which contains DNA sequence integrated into genom of yeast cell and coding the fragment of amino-acid sequence from position 86 to 607 of core protein of hepatitis E virus of genotype 3 (rtHEV-ORF2) under control of promoter of MOX gene. The method allows obtaining immunogenic antigene of hepatitis E virus, which has properties of natural protein. Based on the obtained antigene there created is recombinant vaccine for prophylaxis of hepatitis E virus. Vaccine includes effective amount of rtHEV-ORF2 protein, adjuvant and a physically acceptable diluter. |
 Membrane protein renaturation method / 2500684Renatured membrane protein obtaining method is proposed. The above method involves production of a homogeneous solution containing a denatured membrane protein, a detergent mixture, phospholipide or phospholipide mixture and apolipoprotein or its equivalent with: further removal of detergents and formation of lipid-protein nanodiscs (LPND) containing renatured protein. |
 Extraction method of recombinant proteins / 2499052Invention proposes a production method of recombinant protein through its hybrid precursor substance with natural decomposition site with enteropeptidase. The result is achieved by replacement in natural decomposition site with enteropeptidase Asp-Asp-Asp-Asp-Lys of amino-acid residue of lysine (Lys) with amino-acid residue of arginine (Arg) and further decomposition of hybrid precursor substance with light catalytic subunit of enteropeptidase of a human being or a bull. |
 Hybrid insecticide protein, nucleic acid molecule coding said protein, transgenic plants and seeds thereof containing said protein, method of producing protein and use thereof / 2497830Present inventions relate to protein engineering, plant molecular biology and pest control, as well as a hybrid insecticide protein and use thereof. Described is a hybrid insecticide protein which includes from the N-end to the C-end an N-end portion of Cry3A protein which is fused with the C-end portion of Cry1Ab protein, wherein the position of the crossover of the Cry3A protein and the Cry1Ab protein is located in a conservative block 2, in a conservative block 3 or in a conservative block 4 and has anti-western corn rootworm activity. Also disclosed are nucleic acid molecules which code the novel proteins, methods of producing proteins, methods for use thereof, as well as transgenic plants and seeds thereof which contain such proteins. |
 Method of production of recombinant antigen g2 of hantavirus dobrava in cells of e coli / 2495938Method is characterised in that the DNA of the structure RNAb indicated on Figure 1, which encodes the fused protein of three parts, where N-terminal position is green fluorescent protein GFP, central - peptide of 73 amino acid residues with the amino acid sequence of SRKKCNFATTPICEYDGNMVSGYKKVMATIDSFQAFNTSYIHYTDEQIEW KDPDGMLKDHLNILVTKDIDFDT, and C-terminal - light chain of double-stranded protein Kunitz-type inhibitor from potato tubers (PKPI-BI), are introduced into cells of E. coli. The cells transformed by this construction are cultured, the biomass is lysed, the insoluble fraction of the lysate is separated by centrifugation. The product of expression in the form of inclusion bodies is solubilised with the denaturant. Chromatography is carried out under denaturing conditions. The resulting product is used for detection of specific antibodies in serum of patients with hemorrhagic fever with renal syndrome. |
 Cell for production of heteroproteins and method of production on its basis / 2494148Method is proposed to produce a polypeptide, including cell cultivation, which intensely expresses a bicarbonate carrier and has a transferred DNA, which codes the desired polypeptide. |
| Glarea lozoyensis mutant strain and its application / 2507252 Mutant strain is obtained by impact on Glarea lozoyensis ATCC 20957 strain by nitrosoguanidine and it is deposited in CGMCC with number CGMCC 2933. |
FIELD: chemistry.
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.
4 ex
The technical field
The invention relates to biotechnology and pharmacology and the way of obtaining food by microbial protein synthesis. Most effectively present invention can be used in food and medical industries.
The level of technology
Traditional sources of mushroom protein in the human diet are the fruiting bodies of strains higher edible fungi genera Agaricus, Flammulina, Lentinula, Morchella, Panus, Pholiota, Pleurotus, Volvariella etc. produced by the method of solid-phase cultivation. The use of industrial methods of obtaining fungal biomass by microbiological synthesis contributes not only to intensify the process of cultivation of bacteria of higher edible mushrooms, but also extends the range of potential producers.
A method of obtaining protein biomass strains of Pleurotus ostreatus ACIM F - 697 by the method of deep cultivation in conditions of aeration on a nutrient medium with the subsequent stages of separation and drying of biomass [EN 2092559, SR 21/00, C12N 1/14, C12N 1/14, C12R1:645, publ. 10.10.1997]. Fermentation is carried out in pH range from 6.0 to 7.5 and temperature of 26 to 28 degrees With the conditions of aeration (0,5 l of air/l environment in minutes) at different nutrient medium with the addition of 0.1 to 3% of surfactants (bone fat or vegetable oil - sunflower, corn) by volume. Seed pre "sagalaeva" at a temperature of 4 to 6 C over from 4 to 24 h, which, in authors ' opinion, allows to reduce significantly the duration of fermentation by obtaining multiple peaks in the dynamics of growth of culture. Department of biomass carried out by filtering cultural liquid. The fermentation process takes 24 to 48 hours Yield of dry biomass is from 13 to 23 g/l, the mass fraction of protein - from 20 to 50% on AFM. The disadvantage of this method is a long way to prepare seed material by the method of "zachelacivania", and also the use of an additional component of the nutrient medium - surfactant (bone fat or vegetable oil - sunflower, corn), which increases the process of biomass production and the regularity of the process that is technologically inefficient.
A method of obtaining protein biomass strains of Pleurotus ostreatus WKMR-720 [EN 2126835, SR 21/00, 12N 1/14, 12R 1:645, publ. 27.02.1999] in underlying conditions, based detachable-topping-up method. Duration of one cycle is 30 to 35 hours, with a volume of selected cultural liquid is from 40 to 50% of the total. The duration of the process is 2.5 to 3 months. Seed pre "sagalaeva" at a temperature from 4 to 12 OC for 4 to 8 hours, fermentation is carried out in pH range from 6.0 to 7.5 and temperature of 26 to 28 degrees With the conditions of aeration (0.5 l/l/min) at different nutrient medium with the addition of 0.1-1% of surfactants by volume. The yield of dry biomass is from 13 to 20 g/l, the mass fraction of protein - from 20 to 50% on AFM. The disadvantage of this method is relatively low productivity strain biomass, as well as the use of expensive surfactants.
There are ways to get protein biomass strains of Pleurotus ostreatus 2-204 ACIM F-811 [EN 2189395, SR 21/00, C12N 1/14, C12N 1/14, C12R1:645, publ. 20.09.2002] and strain Panus tigrinus 3-204 ACIM F-810 [EN 2186851, SR 21/00, C12N 1/14, C12N 1/14, C12R1:645, publ. 10.08.2002] deep conditions detachable-topping-up method with subsequent stages Department of biomass by filtering cultural liquid and drying of biomass at a temperature of 60 degrees C. the Duration of one cycle is 60 hours using weaning cultural liquid in the amount of 40 to 50% of the total. Fermentation is carried out in pH range from 5.8 to 6.2 and temperatures from 26 to 28 C and 32 to 34°for strains of Pleurotus ostreatus and Panus tigrinus, respectively, in the conditions of aeration (from 0.8 to 1.0 l/l/min) and mixing on a nutrient medium, containing dry or native whey from 80 to 100 g/l, (NH 4 ) 2 SO 4 - 0,250 g/l or (NH 4 ) 2 HPO 4 - 0.125 g/l. At the initial stage for 18 to 20 hours of cultivation strains as antifoam used sterile unrefined sunflower oil in a concentration of 0.05% vol. to the working volume of the device. The yield of dry biomass ranges from 17 to 25 g/l, the mass fraction of protein for strains of Pleurotus ostreatus and Panus tigrinus - from 28 to 40% and 36 to 40% on AFM respectively. The disadvantage of these methods is the duration of the process of cultivation of strains, and relatively small value of a mass fraction of a crude protein in biomass.
A method of obtaining protein biomass of food and fodder strains of Penicillium nonatum and Penicillium chrysogenum [US 3865951; C12j 13/06, A23j 3/00, publ. 11.02.1975] by the method of deep cultivation and subsequent stages of separation of biomass from cultural liquid, washing biomass, filtering, drying. As the substrate for cultivation of strains-producers assume the use of the various plant materials, such as feed wheat, hydrolysates potato, sugar beet molasses, bagasse and/or citrus waste. Cultivation is expected to be implemented in the temperature range from 25 to 35 C, preferably around 30 C and in the pH range from 4.0 to 7.0, maintaining the pH value at the level corresponding to the maximal growth rate. The number of seed fermenting is 5 to 10% by volume, preferably 10%. Appropriate entry in the nutrient medium of vitamins (such as Biotin) to ensure maximum growth rate of culture, and also non-toxic agent. The duration of the periodic process of cultivation in the conditions of aeration and mixing is from 20 to 48 hours in the case of application of lactose as carbon substrate time of cultivation can be increased up to 72 hours. The process is limited by the basic substratum. Mass fraction of protein in biomass is obtained from 43 to 47% (of total nitrogen on Kjeldal from for 6.81 to 7,53%, the conversion factor -6,25). The disadvantage of this method is relatively low content of protein substances received in biomass.
A method of obtaining protein biomass strain Neurospora sitophila [US4938972, A23L 1/00; publ. 03.07.1990; Moo-Young, 1993. Fermentation of cellulosic materials to mycoprotein foods/ Murray Moo-Young, Yusuf Chisti, Dagmar Vlach // Biotech Adv. - 1993. - Vol.11, - pp.469-479] by the method of deep cultivation of various crops waste subjected preliminary alkaline hydrolysis. The cultivation is carried out in a range of pH 5.5 to 7.5 and temperature from 20 to 40 C, preferably at 26 C, in conditions of aeration (from 0.5 to 1.0 l/l/min) and mixing. The required content of dissolved oxygen in the environment is about 50%, a level below 30% leads to a sharp decrease in productivity. The quantity of seed is from 3 to 10% of working volume of the fermenter. The process is carried out periodic and continuous way. The most effective for this process are devices with relatively low intensity of mixing (arletty bioreactor)than units with mechanical mixing. The end of the process is determined by the achievement of culture stationary phase of growth. Mass fraction of protein in biomass ranges from 30 to 60% for AFM. The disadvantage of this method is the low productivity protein substances - from 2.2 to 2.6 g/L.
A method of obtaining food mycelium biomasses strains kind Polyporus. Polyporus squamosus and Polyporus brumalis [US 4212947, C12N 1/14, C12R 1/645, publ. 10.07.1980] by the method of deep cultivation and subsequent stages of allocation of biomass by filtering (filter press or vacuum filters) or separation and spray drying of biomass. The cultivation is carried out detachable-topping-up method at the temperature from 24 to 28 C, pH of 6.0 to 7.0, aeration from 0.6 to 1.0 l/l/min on a nutrient medium of the following composition,% Mas.: molasses is from 4 to 5%, NH 4 NO 3 - 0,2%, KN 2 RO 4 - 0,12%, vegetable oil - to 0.04% as antifoam. The concentration of inoculum is 10%. The duration of the first cycle is 24 hours, the subsequent cycles of 5 to 7 h with weaning cultural liquid in the amount of 50% of the total. The duration is from 1 to 5 days. Biomass yield - 9 g/l AFM. Mass fraction of protein - 50 to 60%. The disadvantage of this method is negligible accumulation of biomass, which leads to an increase in the number of production cycles and higher costs of water.
A method of obtaining food mycelium biomass of fungi of the genus Fusarium: strains of Fusarium species frost, Fusarium oxyspomm and Fusarium solani [US 4501765, A23J 3/00, publ. 26.02.1985; US 4555485, SR 21/00, C12R 1/77, publ. 26.11.1985] by the method of deep cultivation and subsequent stages of separation of biomass from cultural liquid, washing, filtration, denuklearizatsii biomass drying. As the substrate for cultivation of strains-producers assume the use of different carbon-containing raw materials (starch, starch-containing raw materials, as well as the products of hydrolysis, sucrose, saharoponijatee raw materials and their hydrolysates (invert sugar etc), hydrolyzed potatoes, molasses, glucose, maltose, hydrolyzed starch legumes or cassava). The cultivation is carried out in the temperature range from 25 to 34 C, preferably 30 C and the amount of seed fermenting from 5 to 10% by volume, in the range of pH of 3.5 to 7.0, it is preferable to maintain a constant corresponding to the maximal growth rate of culture. Preferably the introduction into a nutrient medium of vitamins (such as Biotin) to ensure maximum growth rate of culture, and also non-toxic agent. The process is carried out periodic [US 4501765, A23J 3/00, publ. 26.02.1985] and continuous [US 4501765, A23J 3/00, publ. 26.02.1985; US 4555485, SR 21/00, C12R 1/77, publ. 26.11.1985] way. The duration of the periodic process of cultivation in the conditions of aeration and mixing is from 20 to 48 hours. The process is limited by the basic substratum. Mass fraction of protein in the resulting biomass is between 45 and 61% (of total nitrogen on Kjeldal - from 7.2 to 9.9%, the conversion rate - 6,25). The process of reducing the RNA in biomass carried out by exposure to solvents (lower alcohols containing not more than 3 carbon atoms) at a concentration of 40 to 100% within 1.5 to 40 minutes at a temperature of 45 to 60 degrees C [US 4501765, A23J 3/00, publ. 26.02.1985] or by a sharp biomass heating up to a temperature of over 68 C, preferably from 72 to 74 C, for 30 to 45 min [US 5739030, C12N 1/14, publ. 14.04.1998]. The method allows to reduce the amount of RNA to less than 2%, but the loss of biomass in this range from 30%to 33%. The disadvantage of this method is the high content in the biomass RNA (from 7 to 12%), which significantly reduces acceptable for human consumption the amount of biomass (not more than 2 g / day) and requires stage of denuklearizatsii that, in turn, leads to loss of biomass (from 30% to 33% AFM).
Thus, at present, no known method for obtaining edible mushroom biomass the method of depth of cultivation, providing high speed the growth of the high-protein accumulation of biomass with high concentration of valuable unsaturated fatty acids and low concentration of nucleic acids (from 3.0 to 4.1% AFM), which allows to use this biomass for human nutrition.
Disclosure of the invention
The aim of the invention is development of a method of obtaining food
the biomass of fungi with high protein content (from 53 to 63% AFM) and the low number of nucleic acids (from 3.1 to 4.1% AFM) by the method of deep cultivation with high speed of growth.
The method of obtaining food protein biomass is characterized by the fact that, as the producer strain used a strain of Fusarium sambucinum Fuck. var. ossicolum (Berk. et Curt.) D-104 deposited in the PMBC number F-1161, the cultivation process is carried out in-depth conditions in the range of pH of 3.5 to 7.0 in conditions of aeration (from 0.5 to 2.0 l/l/min with a line speed of the mixing device from 1 to 4.9 m/s). As a source of carbon use different carbon-containing raw materials (products of various kinds of grain (wheat, rye, triticale), potatoes, sugar beet and cane juice roots and green part of Jerusalem artichoke, turnips, beetroot, and other species and their hydrolysates) in the amount of 2.0 to 6.0%by weight.
Temperature the process of cultivation in each cycle fermentation support from the beginning of the loop to the switching point at the level of 26 to 30 C and the next-to-end cycle - at the level of 22 to 25 C, with the switching point is determined by the accumulation of biomass to the concentration of 45 to 60% of the maximum achievable in this unit (maximum achievable concentration of biomass depends on the mass transfer characteristics of the fermenter, which hosts the cultivation of microorganism and its specific value is determined experimentally-based trial downloads).
In another embodiment, the method switching point determines the concentration of dissolved oxygen after its reduction to the amount from 20 to 40% from saturation (in terms of atmospheric air pressure).
The process is cyclical. In the first cycle quantity of seed is from 1 to 5.%. In subsequent cycles weaning cultural liquid of the vehicle is from 95 to 99%.
Deep cultivation was carried out in periodic mode in a fermentation apparatus 30 litres displacement of 20L, pH statorvane when set to 4.0, the excessive pressure 0,4 MPa, aeration in sterile conditions 1.0 l/l/min with mechanical agitation 400 rpm mixer turbine type on a nutrient medium of the following composition: wheat starch - 45,0 g/litre; the ammonium nitrate - 4.5 g/l; potassium phosphate one-deputizing - 1.8 g/l; corn extract condensed - 10.0 g/l, propanol B-400 - 0.4 g/L. Starch was exposed to the primary enzyme hydrolysis drug amilosubtilin in the calculation of 0,024% to volume of a medium (4.8 g/l). Wednesday was inocularea mycelial suspension (second and subsequent passages) in the amount of 3% by volume. Pre-defined maximum achievable on the environment for producer strain concentration of biomass at the end of fermentation cycle, which was 2 to 5 g/l AFM. The temperature regime of the cultivation process was supported from the beginning of the loop to the switching point at the level of 26 to 30 C, next to the end of the cycle - at the level of 22 to 25 C, with the switching point was determined by the accumulation of biomass to the concentration of 11.0 g/l AFM, which accounted for about 45% of the maximum concentration in this unit. Dehydration cultural liquid carried out by suction filter, drying of wet biomass were performed in the fluidized bed dryer.
The chemical composition of the biomass, % to the AFM:
Total protein - 58,0;
Lipids - 7,8;
Nucleic acid - 3,1.
Example 2
Cultivation producer strain were performed similarly outlined in example 1, however, the switching point was determined by the accumulation of biomass to the concentration of 15.0 g/l AFM, which corresponded to 60% of the maximum concentration in this unit. Dehydration cultural liquid carried out by suction filter, drying of wet biomass were performed in the fluidized bed dryer.
The chemical composition of the biomass, % to the AFM:
Total protein - 62,1;
Lipids - 6,5;
Nucleic acid is 3.7.
Example 3
Cultivation producer strain were performed similarly outlined in example 1, however, the switching point was determined by the concentration of dissolved oxygen after its reduction to the amount of 20% of saturation (in terms of atmospheric air pressure). Dehydration cultural liquid carried out on the drum vacuum filter, drying of wet biomass were performed in the fluidized bed dryer.
The chemical composition of the biomass, % to the AFM:
Total protein - 63,0;
Lipids - 6,4;
Nucleic acid - 4,0.
Example 4
Cultivation producer strain were performed similarly outlined in example 1, however, the switching point was determined by the concentration of dissolved oxygen after its reduction to the value of 40% from saturation (in terms of atmospheric air pressure). Dehydration cultural liquid carried out on the drum vacuum filter, drying of wet biomass were performed in the fluidized bed dryer.
The chemical composition of the biomass, % to the AFM:
Total protein - 60,8;
Lipids - 7,0;
Nucleic acids and 3.3.
Thus, the use of this method allows to get high-quality mushroom protein biomass food purposes with low content of nucleic acids (from 3.1 to 4.1% AFM)that it requires additional stage of denuklearizatsii biomass and therefore avoids unwanted products splitting NC. The method can be successfully used in food industry and pharmacology for the production of edible mushroom protein biomass.
1. The method of obtaining food fungal biomass with high content of protein, providing cyclical deep cultivation producer strain liquid the nutrient medium, containing sources of carbon, nitrogen, mineral salts, separation and drying of wet biomass of fungus, notable as a producer of biomass use a strain of the fungus Fusarium sambucinum ACIM F-1161, the cultivation process is carried out at pH of 3.5 to 7.0 in conditions aeration from 0.5 to 2.0 l/l/min, the temperature regime of the cultivation process in each cycle fermentation support from the beginning of the loop to the switching point at the level of 26 to 30 C and the next-to-end cycle - at the level of 22 to 25 C, with the switching point is determined by the accumulation of biomass to concentration from 45 to 60% of the maximum achievable in a fermentation apparatus or switching point determines the concentration of dissolved oxygen after its reduction to the amount from 20 to 40% from saturation (in terms of atmospheric air pressure).