The strain streptomyces hygroscopicus 7 - producer proteolytic enzyme hygromycin
(57) Abstract:The invention relates to biotechnology and can be applied in the leather industry and veterinary medicine. Strain the received selection by under the influence of mitomycin C and stored at the research Institute of agricultural Microbiology at number ARRIAM-481 and in the group alipanah microorganisms. The level of productivity of strain 7 is increased and ranges from 120-170 PE/ml The present invention relates to medical and microbiological industry and can be used to produce proteolytic enzymes.The objective of this technical solution was to obtain a new strain with higher performance to obtain hygromycin, which can be used in veterinary practice for the treatment of dyspepsia animals in combination with antibiotics as growth biostimulant animals and birds, with the addition of it in the feed, in the leather industry for dehairing and bating of raw hides, as well as for protein hydrolysis of different origin.Hygromycin can be used in medicine when nonspecific respiratory diseases (rasla lizirovania and rejection scab, lysis of purulent surgical fibrinous masses and diseases.During solution was selected strain with high ability to synthesize hygromycin.A method of obtaining a proteolytic enzyme hygromycin using as a producer of culture of Actinomyces hygroscopicus (SU, author's certificate N 663715, CL C 12 D 9/14, C 12 D 13/10. Published 25.05.79. Bulletin No. 19). However, the proteolytic activity of Actinomyces hygroscopicus in a specified way reaches only 35 PE/ml. the Disadvantage of this technical solution is low proteolytic activity in the culture fluid.The closest to the technological nature of the invention is a strain of Streptomyces lavendulae PMBC S-910 - producer of proteolytic enzyme activity in the culture fluid by Kunitz 40-97 PE/ml, which is chosen for the prototype. (SU, author's certificate N 1735364 class. C 12 N 9/52, 1/20, Patent USSR N 1001862, CL C 12 N 9/48, 1983). The lack of strain is low proteolytic activity.The proposed strain Sreptomyces hygroscopicus 7 obtained by the method of selection for the effects of mitomycin C at a dose of 100 μg/ml Strain produces up to 170 PE/ml of culture fluid. The strain according to the invention is stored in Nauch microorganisms.The essential features of the claimed strain of Streptomyces hygroscopicus 7, in common with the prototype.Strain peptonized milk, gelatin liquefies, does not form hydrogen sulfide, nitrates does not recover. As a source of carbon assimilates glucose, Inositol; weakly increases on the environment arabinose. On potato forms active growth. The proposed strain as well as the prototype, does not form a soluble melanoidin pigments on all agar and liquid media used for cultivation.Distinctive features of the proposed strain from the known (prototype)
The proposed strain unlike the prototype Streptomyces lavendulae PMBC S-910 refers to the species Streptomyces hygroscopicus and all of the morphological and cultural characteristics different from the prototype.In the medium with corn extract and starch (N 21/12) at 21 days of growth at 251oC forms a colony 8-9 mm flat with pleats in the center and shyrokoradiuk jagged edge. Aerial mycelium velvety, moderately developed, on the edge of the colony is partially reduced in the form of a fringe of snow-white color (d-3). Substrate mycelium buckskin (1). The medium is not colored. On the organic environment N 2 (Gause) colonies are convex, diameter 8-11 mm with knob in the center and flat izraza is painted.On ovsanna agar N 61 colonies large 13-16 mm, convex with shyrokoradiuk rough edges. Aerial mycelium velvety, richly sorulari dark ash (4), on the edge of a colony of white color. Substrate mycelium pale sand (3). The medium is not colored.On agar of čapek with starch forms a sparse growth of colonies of small, 2-3 mm wide, flat, with rugged shyrokoradiuk edge. The aerial mycelium of powdery, smoky (l 1), abundantly sorulari covers the colony completely. Substrate mycelium gray (4). The medium is not colored.On agar medium of Tresner forms a meager growth, colonies 2-3 mm, convex with tubercle in the centre and jagged edge. The aerial mycelium of powdery, white color (l 3), respirology. Substrate mycelium yellowish brown (d-4). The medium is not colored.Unlike the prototype does not grow on medium containing fructose, well metabolizes xylose, moderately growing on medium containing sucrose. The inventive strain exceeds the level of biosynthesis of proteolytic enzyme complex for 75% of the prototype (the claimed strain 120-170 PE/ml; prototype - 40-97 PE/ml).Morphological and cultural characteristics
In the medium with corn extract and starch (N 21/12) for 21 days rodusky mycelium velvety, developed moderately, on the edge of the colony is partially reduced in the form of a fringe of snow-white color (d-3). Substrate mycelium buckskin (1). The medium is not colored. On the organic environment N 2 (Gause) colonies are convex, diameter 8-11 mm with knob in the center and flat jagged edge. Aerial mycelium velvety, white (d-3). Substrate mycelium yellowish brown (d-4). The medium is not colored.On ovsanna agar N 61 colonies large 13-16 mm, convex with shyrokoradiuk rough edges. Aerial mycelium velvety, richly sorulari, dark ash (4), on the edge of a colony of white color. Substrate mycelium pale sand (3). The medium is not colored.On agar of čapek with starch (N 84) forms a sparse growth of colonies of small, 2-3 mm wide, flat, with rugged shyrokoradiuk edge. The aerial mycelium of powdery, smoky (l 1), abundantly sorulari covers the colony completely. Substrate mycelium gray (4). The medium is not colored.On agar medium of Tresner (N 64) forms a meager growth, colonies 2-3 mm, convex with tubercle in the centre and jagged edge. The aerial mycelium of powdery, white color (l 3), respirometry. Substrate mycelium yellowish brown (d-4). The medium is not colored.Physio is on Tue 9th days of growth at 28oC, H2S does not form on the tissue grows moderately, on the potato produces abundant growth grayish color, does not restore the nitrate to nitrite, melanoidin pigments are not formed. Response to inversion of sucrose negative.On the environment Pridham-Gottlieb grows well in the presence of glucose, maltose, xylose, grows moderately in the medium with Na-citrate, Inositol, sucrose, sorbitol, weakly increases with arabinose, lactose, Ramezay, galactose, mannitol. Does not grow on medium containing fructose and acetic acid sodium.The temperature optimum for growth and development of the 28oC, the minimum temperature for growth of 20-25oC, can withstand a maximum temperature of 32oC.Storage conditions of the proposed strain
The strain is stored on agar 21/12 with starch and corn extract at room temperature for 3 months.The composition of the medium 21/12:
1. Corn extract 5 g (dry weight)
2. (NH4)2HPO44 grams
3. KH2PO42 g
4. MgSO47H2O 0.25 g
5. CaCO31 g
6. The soluble starch 20 g
7. Agar-agar 25 g
8. Tap water to 1 liter
the pH of the Medium prior to sterilization 7,3
Sterilization 40 min to 0.8 MPa.Anta is SS="ptx2">The inventive strain is not toxic and does not apply to microorganisms with pathogenic properties.In submerged fermentation declare strain accumulates in the culture liquid of the proteolytic enzyme action - hygromycin.Conditions of education hygromycin
To obtain hygromycin strain cultured for 72-96 h on a rocking chair (speed 220-250 min-1in flasks of Erlenmeyer with a capacity of 750 ml, containing 50 ml of medium of the following composition, %:
protein-vitamin concentrate (PVC) - 3,0
corn flour - 2,0
gidrol or green syrup - 6,0-7,0
chalk - 1,2
potassium phosphate one-deputizing - 0.02
sunflower oil - 1,0
water tap - rest
Bulb inoculant 5 ml of inoculum, obtained by cultivation in flasks of Erlenmeyer with a capacity of 750 ml (281oC, 220-250 rpm) containing 100 ml of medium of the following composition, %. soy flour - 1,5, corn extract to 0.2, (NH4)2SO4to 0.2, NaCl and 0.5, CaCO3to 0.3, glucose 2%, and the BVK is 1.0. The period of growing seed 48 hoursFor preparation of inoculum used cultures grown in test tubes with the medium 21/12. The culture was grown in flow. , Preparation and assay of enzymes. 2. Chymotrypsinogenes and chymotrypsins. Methods Enzymol., 1955, v.2, p. 8-26). The strain Streptomyces hygroscopicus 7 - producer proteolytic enzyme hygromycin.
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
FIELD: microbiological industry.
SUBSTANCE: invention relates to nutrient media used for culturing producers of carotene. Invention proposes nutrient medium containing barley flour, soybean meal, potassium dihydrogen phosphate, sunflower oil, vitamin B1, β-ionone and tap water being components are taken in the following ratio, wt.-%: barley flour, 3.0-4.0; soybean meal, 4.0-4.7; sunflower oil, 3.8-4.0; potassium dihydrogen phosphate, 0.04-0.05; vitamin B1, 0.0002-0.0005; β-ionone, 0.098-0.099, and tap water, the balance. The proposed nutrient medium is low-priced and enhances the biosynthetic ability of producer of carotene.
EFFECT: valuable properties of nutrient medium.
3 tbl, 3 ex
FIELD: biotechnology, microbiology.
SUBSTANCE: invention relates to producing ethyl alcohol or fodder for monogastral animals. The polyenzyme product with glucoamylase, proteolytic and xylanase activities is prepared by fermentation of wheat bran using microorganism Aspergillus niger. Indicated glucoamylase, proteolytic and xylanase activities have the following minimal values: glucoamylase activity - at least 100 U/g of dry matter; proteolytic activity - at least 100 U/g of dry matter; xylanase activity - at least 100 U/g of dry matter under condition that glucoamylase activity has to be at least 750 U/g of dry matter and/or xylanase activity has to be at least 300 U/g of dry matter. Invention provides enhancing the soluble nitrogen content in wort after saccharification, reducing viscosity and effectiveness in using.
EFFECT: improved preparing method, valuable properties of hydrolyzed bran.
14 cl, 10 tbl, 7 ex
FIELD: biotechnology, microbiology.
SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.
EFFECT: improved preparing method.
15 cl, 4 ex
FIELD: agriculture, fruit storage.
SUBSTANCE: the present innovation deals with technology to treat fruit before their laying for storage. For the purpose to decrease the loss from microbial spoiling and increase the terms of storage fruit essential oils and epiphytic substances of cuticular layer of plant raw material should be applied onto the surface from mycella obtained after extracting Saprolegnia parasitica micromycete biomass with nonpolar extragent in supracritical state.
EFFECT: higher efficiency of protection.
FIELD: food industry, confectionary industry.
SUBSTANCE: the suggested concentrate should be prepared due to pressing the juice under aseptic conditions out of sugar beet prepared, deodorated and sterilized with carbon dioxide in supracritical state in the field of ultrasound fluctuations, cultivating upon residues of mycellial fungi of Trichoderma and Aspergillus species of citric acid fermentation, separating culture liquid, its mixing with juice and introducing liquid ammonia into the blend along with supracritical CO2-extract out of Mortierella nigrescens micromycete biomass further extracted according to the preset technique to obtain a solid residue treated with liquid ammonia followed by concentrating the blend, treatment of concentrate with liquid carbon dioxide, mixing Mortierella nigrescens micromycete biomass with a treated solid residue and heating the mixture up to 60 C, not less. The innovation enables to obtain a concentrate of improved structure-forming properties and increased thermal stability.
EFFECT: higher efficiency of manufacturing.
FIELD: food industry, confectionary industry.
SUBSTANCE: the suggested concentrate should be prepared due to pressing the juice out of sugar beet prepared, deodorated with carbon dioxide in supracritical state, extracting residues at mixing water and liquid hydrogen chloride at extractional volume, mixing juice and extract and introducing liquid ammonia into the blend along with supracritical CO2-extract out of Mortierella zychae micromycete biomass further extracted according to the preset technique to obtain a solid residue treated with liquid ammonia followed by concentrating the blend, treatment of concentrate with liquid carbon dioxide, mixing Mortierella zychae micromycete biomass with a treated solid residue and heating the mixture up to 60 C, not less. The innovation enables to obtain a concentrate of improved structure-forming properties and increased thermal stability.
EFFECT: higher efficiency of manufacturing.
FIELD: microbiology, biotechnology.
SUBSTANCE: invention proposes the strain Mucor racemosus № 195 for preparing immunogenic preparations used against mucorosis in agricultural animals. This strain elicits the high spore-forming capacity, absence of pathogenicity and it produces polysaccharides eliciting immunogenic properties.
EFFECT: valuable properties of strain.
2 tbl, 2 ex
FIELD: biotechnology and agriculture, in particular fungi production.
SUBSTANCE: claimed method includes preparation of mycelium biomass on nutrient medium in presence of growth stimulator (e.g., Azospirillum bacterium suspension) and seeding of mycelium biomass on cereal nutrient medium. Mycelium biomass is prepared by deep cultivation; and as nutrient medium potato-wheal medium is used.
EFFECT: accelerated method for production of edible fungal seed mycelium.
FIELD: food industry.
SUBSTANCE: prepared sugar beet should be deodorated and sterilized with supercritical carbon dioxide in the field of ultrasound fluctuations, juice pressing should be carried out under aseptic conditions followed by cultivation upon residues of mycellar fungi of Trichoderma and Aspergillus species of acetic acid fermentation. Then comes separation of culture liquid, its blending with juice, addition of ammonia and supercritical CO2-extract into the blend out of Mortierella reticulata micromycete biomass extracted then according to the preset technique to obtain solid residue treated with liquid ammonia followed by concentrating the blend, treating the concentrate with liquid carbon dioxide, mixing with solid residue-treated Mortierella reticulata micromycete biomass and heating the mixture up to 60 C, not less. The innovation provides improved structure-forming capacity and increased thermal stability of gelling concentrate.
EFFECT: higher efficiency.