Strain of micromycete aspergillus oryzae-producer of proteolytic and amylolytic ferments to be used in food industry
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
The invention relates to biotechnology, in particular the production of strain - producer of proteolytic and amylolytic enzymes, and can be used for hydrolysis of vegetable and animal substances in food industry, fermentation industry, agriculture.
It is known that the filamentous fungus Aspergillus oryzae has wide application in the food industry, as they do not have pathogenic properties. Biosynthesis of proteases and amylases, characteristic for representatives of Aspergillus oryzae, used, for example, in the manufacture of certain food products (fermented seasonings, fermented drinks)when you want the hydrolysis of proteins and carbohydrates (EN 2361914, 2009, EN 2187947, 2002).
A known strain of Aspergillus oryzae VKPM F-369, with the ability to produce complex proteases exhibiting maximum activity in the acidic and weakly acidic zone pH, α-amylase. However, the main disadvantage of this strain is the lack of synthesis of related hydrolases, as well as the long period of growth and development of micromycete; strain intensively produces α-amylase and protease in liquid media only on the fourth - fifth day (SU 1440922, 1988).
A known strain of Aspergillus oryzae VKPM F-683, producing complex acidic and weakly acidic protease, α-amylase and related xylanase, glucanase and cites. It is known that deep cultivated and this strain produces a complex of highly active enzymes. The resulting enzyme complexes used in the food industry.
However, the application of strain is limited due to a lack of high growth rate and specific requirements for nutrient environments, as it is growing on media with solids content of not more than 18% (EN 2070921, 1996).
The task of the invention consists in obtaining a strain of Aspergillus oryzae producing protease and α-amylase with high growth rate, mainly for solid-phase cultivation.
As the invention proposes a new strain of micromycete Aspergillus oryzae 37-53, obtained by the method of multi-stage selection and mutagenesis deposited in the GSI research Institute for agricultural Microbiology in the Departmental collection of useful microorganisms for agricultural purposes RAAS (RCAM) under registration number RCAM01135. The strain is a producer of protease and α-amylase, has a high growth rate, mainly for solid-phase cultivation and intensive sporoobrazovanie.
The technical result of the invention is a higher growth rate of the proposed strain of Aspergillus oryzae RCAM01135.
In particular, in environments with soybean meal highs proteoliticheskoi and amylolytic activities occur on the third day of cultivation, and the intensity of sporoobrazovanie was the most high is also some on the third day. This allows to consider the claimed strain as a promising tool for use in the food industry for processing of agricultural raw materials, such as soybean, with the aim of obtaining various food flavoring additives, fermentation production, the baking industry.
A strain of Aspergillus oryzae RCAM01135 characterized by the following cultural-morphological and physiological-biochemical characteristics.
Cultural morphological traits:
(a) the nature of growth (pubescence, the edge of the colony). Colony fluffy, rounded, smooth edge, white mycelium, spore-forming, colony color from olive to dark green. Vegetative mycelium septate, well-branched with large swellings, the thickness of the hyphae 6-12 µm; conidia are formed exogenously, the surface of conidia smooth, rounded form, the diameter of the spores 5-6 μm; forms a conidial heads with single-stage sterigmate;
b) the size of the colony (at a certain time of incubation). 3 days of growth on wort-agar the colony diameter 50×51 mm;
b) aerial mycelium (availability, color, features of sporulation). Plentiful sporification dark green color. On the edge of the colony white fluffy mycelium 3 mm;
g) the color of the substrate mycelium. White;
d) the color of the reverse side of the colony. Light brown;
e) other signs (pigment, the exudate is so). The pigments are not, exudate absent;
g) changing the culture as we grow older. The color of the colony darkens with age, becoming brownish-green;
C) behavior in other environments. On Chapek's medium with glucose: 3 day growth colony color - green, surface smooth; the diameter of the colony 32×34 mm
Studies of the morphological characteristics of the culture identification of strain in accordance with the determinant of filamentous fungi (Bilai VI, Koval AS Aspergilli. The determinant. - Kiev: Nauk. Dumka, 1988, - 204 C.).
Physiological and biochemical characteristics
Relation to oxygen - aerobe. Optimum growth temperature is 28-30°C. the Maximum temperature is 50°C. the Minimum temperature is 18°C. the Optimum pH value for growth of the fungus is 5.5. Fungus growth is observed in the pH range from 2.5 to 10.0.
As a source of carbon to the fungus uses starch, glucose, sucrose, xylose, maltose, mannitol, glycerol and lactose. Assimilates nitrate, ammonium and amine nitrogen, proteins. The strain is not pathogenic. In the mode of deep and solid state cultivation was established ability to synthesis of enzymes, amylolytic and proteolytic action.
The strain is stored on agar malt wort 8% CB; pH environment - natural; storage temperature +20-25°C.
Deep and solid phase kultivirovala strain Aspergillus ryzae RCAM01135
Deep cultivation of the strain Aspergillus oryzae 37-53 was performed on various complex natural environments tailored to the physiological needs of the producer. When designing nutrient media took into account the fact that for the synthesis of protease and α-amylase favorable starch, proteins, therefore, the main components of the environment was grain raw materials. As a source of phosphate nutrition for micromycete applied potassium phosphate. Thus, for submerged cultivation of a strain of Aspergillus oryzae RCAM01135 were used nutrient medium of the following composition, %:
1. Barley flour - 3,0, wheat bran - 3.0, starch - 2,0, KH2PO4- 1,5;
2. Wheat flour is 6.0, bran - 2,0, KH2PO4- 1,5%;
3. Soy flour - 6,0, barley flour - 5,0, KH2PO4- 1,5;
4. Soy flour is 6.0, bran - 2,0, KH2PO4- 1,5;
Cultivation of a strain of Aspergillus oryzae RCAM01135 was carried out in Erlenmeyer flasks with a volume of 750 cm3containing 50 cm3sterile nutrient medium, the pie rocking chair with a speed of 220 rpm at 30°C. the results Obtained are shown in table 1.
|Deep cultivated the e fungus Asp.oryzae RCAM01135 natural nutrient media|
|No. environment||Day||Growth (density, color QOL)||pH||RSW, %||Farms, the act of the terrain, units/cm3|
|2||+khaki (yellowish) porridge||4,8||2,8||1,5||0,64|
|3||+khaki (yellow) liquid||are 5.36||2,2||1,8||0,82|
|2||++khaki (yellowish) porridge||5,2||2,0||1,3||0,93|
|3||++khaki light liquid||5,6||2,2||1,5||1,10|
|3||1||+++++very thick beige porridge||5,69||5,0||-||-|
|2||dark beige porridge||7,6||5,0||0,05||0,35|
|3||++++khaki dark porridge||8,46||of 5.4||0,03||0,56|
|4||1||+++++thick beige porridge||7,10||5,0||-||-|
|2||dark beige porridge||8,33||a 4.9||0,06||0,35|
|3||+++dark khaki porridge||8,73||of 5.4||0,02||0,12|
As can be seen from table 1, the selected strain of Aspergillus oryzae RCAM01135 on nutrient media containing barley or wheat flour and wheat bran (environments 1 and 2), showed the ability to synthesis of protease and α-amylase: the total proteolytic activity on the 3rd day of cultivation amounted to 1.5 to 1.8 units PS/cm3, amylolytic 0,82-1,10 unit ° C/cm3. On Wednesday, which consisted soeva flour (medium 3 and 4), the level of activity of synthesized enzymes were decreased in 2-3 times.
Solid-phase cultivation of a strain of Aspergillus oryzae RCAM01135 carried out on wheat bran with humidity 52,4% and soybean meal with humidity to 51.8% at 30°C in stationary conditions. The results are presented in table 2.
|Surface cultivation of the fungus A.oryzae RCAM01135 in various environments|
|The medium for cultivation|
|The characteristic growth of the fungus, the day||Enzymatic activity, units/g|
|Bran||White mycelium, the beginning of sporoobrazovanie, dispute color green||Abundant sporulation, the spores dust, color green with olive tint||Unchanged||43,5||9,0|
|Soybean meal||1||2||3||41,0||to 12.0|
|White mycelium||Abundant sporulation, dispute color green with olive tint||Abundant sporulation, the spores olive green|
The results of in-depth and solid-phase cultivation confirm biosynthetic ability of a selected strain of Aspergillus oryzae RCAM01135 against proteolytic and amylolytic enzymes, which is characteristic of fungi of the genus Aspergillus oryzae species.
Thus, the strain grows well on liquid and solid who's nutrient media, synthesizes enzymes proteolytic and amylolytic activities and identified as the filamentous fungus Aspergillus oryzae.
Cultivation of a strain of Aspergillus oryzae RCAM01135 in environments with soybean meal,
The accumulation of the experimental samples selected culture of Aspergillus oryzae RCAM01135 carried out on medium containing soybean meal and soybean meal with the addition of 10% barley with different humidity for 3 days at 30°C (table 3).
|Biochemical characteristics of experimental samples of the fungus A.oryzae RCAM01135|
|The medium for cultivation|
|Humidity surface. culture %||Biochemical indicators of fungus|
|Protein by Kjeldahl method, % ASV||Protein by Lowry, mg/g ASV|
|With the new meal +10% barley||51,3||41,0||52,8||309,7|
Experimental samples of the fungus A.oryzae RCAM01135 were obtained as a result of rapid growth of micromycete at surface cultivation on soyabean meal, rich in protein, amino nitrogen and can be used for further processing in the production of fermentation products soybeans.
Selected strain of Aspergillus oryzae RCAM01135 and accumulated experimental samples of the fungus A.oryzae RCAM01135 were investigated in a Test lab techno-chemical control and arbitration methods of analysis of the GSI VNIIBT Rosselhozakademii.
The results of the tests for the identification of lags showed that in pure culture micromycete Aspergillus oryzae RCAM01135 transgenic sequence no (test report dated 23 March 2012).
In the analysis of heavy metals in the experimental samples of the fungus A. oryzae RCAM01135 it was found that their content does not exceed maximum permissible concentrations (minutes dated April 18, 2012). Table 4 shows the results of the analysis on the content of heavy metals in the substrate - soybean meal and superficial culture of the fungus A.oryzae RCAM01135. Ana is ulichnye results were obtained in all studied experimental samples.
|The content of heavy metals in surface culture of the fungus A.oryzae RCAM01135|
|The analyzed object||The content of heavy metals, mg/kg|
|Lead Pb||Arsenic As||Cadmium Cd||Mercury Hg|
|(MPC for soybean)1(1SanPiN 220.127.116.118-01 "Hygienic requirements for safety and nutrition value of food products")||0,5||0,3||0,1||0,02|
|Superficial fungal culture A.oryzae 37-53||0,0065||0,0047||0,0029||0,0014|
Strain micromycete Aspergillus oryzae deposited in the GSI ARRIAM (Departmental collection of useful microorganisms for agricultural purposes - RCAM) under registration the first room RCAM01135, producing proteolytic and amylolytic enzymes for use in the food industry.
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.
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SUBSTANCE: invention relates to a strain of the bacteriophage Escherichia coli ECD4. The strain of the bacteriophage Escherichia coli ECD4 is isolated from faeces of broiler chickens on the culture of the bacteria of the strain Escherichia coli O104.H4 RK1No.112027 and is deposited in the State Collection of Pathogenic Microorganisms and Cell Cultures "GKPM-Obolensk" under the number Ph63. The proposed strain of the bacteriophage has lytic activity in respect to the bacteria of the strain Escherichia coli O104:H4 RKINo.112027, lyses Escherichia of the serotype 0157:H7, does not suppress growth of cells Escherichia coli M-17, has lytic activity in respect to several other clinically significant serotypes of Escherichia, and also to several types of Shigella. The strain of the bacteriophage Escherichia coli ECD4 propagates on the laboratory non-pathogenic strain Escherichia coli K-12 C600F.
EFFECT: invention may be used in creation of new preparations for treatment of a disease caused by bacteria Escherichia coli and for elimination of this pathogen from food products.
SUBSTANCE: method is proposed for biodegradation of drotaverine hydrochloride (spasmolytic NOSHPA). The method provides for the process of reaction of drotaverine hydrochloride with cells of the strain Rhodococcus rhodochrous, Institute of Ecology and Genetics of Microorganisms 608. The process is performed under aerobic conditions in the medium containing sources of carbon, nitrogen, phosphorus, mineral salts. At the same time the additional source of carbon in the nutrient medium may be glucose. The cells of the strain Rhodococcus rhodochrous, Institute of Ecology and Genetics of Microorganisms, 608, may be used in the immobilised form. The strain is stored in the Regional Profiled Collection of Alkanotroph Microorganisms (acronym of the collection of the Institute of Ecology and Genetics of Microorganisms). Duration of biodegradation of drotaverine hydrochloride makes at least 80% for 25 days.
EFFECT: invention makes it possible to reduce time of biodestruction of drotaverine hydrochloride and to consider the used strain as a potential agent for cleaning of environment.
4 cl, 4 dwg, 3 ex
SUBSTANCE: method includes preparation of the inoculum of cells Streptomyces sp. MT246 (All-Russian Collection of Microorganisms Ac-2618D). Preparation of the nutrient medium containing one or more sources of carbon, nitrogen, metal ions in the form of soluble salts with fractional feeding of the carbon source during the productive stage. At that, as an additional carbon source the nutrient medium contains rhamnose, and a necessary component of the nutrient medium - MnS04 and sorbent from the group of amberlites XAD. Adding of the inoculate of cells Streptomyces sp. MT246 (All-Russian Collection of Microorganisms Ac-2618D) to the nutrient medium together with rhamnose and MnSO4 in a predetermined ratio and incubating of the medium at pH 7.0-7.5, temperature 25-35°C for 7-10 days. Separation of the residue is carried by centrifugation. The resulting residue is added to ethanol at a predetermined ratio, followed by extraction of tacrolimus at 30 for 2 hours and centrifuging to obtain the extract comprising tacrolimus.
EFFECT: invention enables to increase the yield of tacrolimus.
9 cl, 6 ex
SUBSTANCE: invention relates to biotechnology, and represents a method of producing a recombinant mutein [C112S] of human enterokinase light chain, using a bacterium belonging to the genus Escherichia, transformed with an expression plasmid containing a DNA fragment encoding a mutein [C112S] inactive precursor of human enterokinase light chain, comprising a sequence encoding a cleavable N-terminal peptide comprising a hexahistidine cluster and the enterokinase recognition sequence, and a sequence fused with it in frame encoding a precursor of mutein [C112S] of human enterokinase light chain with non-cleavable C-terminal hexahistidine cluster under control of a promoter operating in the bacterial cell.
EFFECT: invention enables to produce the recombinant mutein of human enterokinase light chain with a high yield.
11 cl, 5 dwg, 1 tbl, 6 ex
SUBSTANCE: invention represents a method of production of precursor of a recombinant fragment of human tissue plasminogen activator (tPA) (reteplase) using a bacterium belonging to the genus Escherichia, transformed with an expression plasmid containing a DNA fragment encoding a precursor of the recombinant fragment of human tissue plasminogen activator (tPA) (reteplase), comprising a sequence encoding a cleavable N-terminal peptide comprising decahistidine cluster and sequence of enterokinase recognition fused in frame, or a fragment of DNA encoding the [-1] methionyl of fragment of human tPA (reteplase), under the control of a promoter operating in the said bacterial cell.
EFFECT: invention enables to obtain a recombinant fragment of human tissue plasminogen activator with a high yield.
10 cl, 6 dwg, 1 tbl, 8 ex
SUBSTANCE: fermentation of nutritional medium is carried out based on corn starch hydrolysates by actinomycete. Then ultrafiltration of the obtained filtrate of the culture liquid is carried out with an activity of 4250±250 IU/cm3 through membranes with cut-off of 5 kDa. The filtrate obtained is concentrated under vacuum of 0.082 mPa at a temperature of 25-30°C. It is clarified at the temperature of 30-40°C. Its ultrafiltration is carried out again through the membrane with cut-off of 1 kDa. The obtained glycosidase inhibitor is dried and ground.
EFFECT: invention enables to obtain the glycosidase inhibitor with increased activity.
1 tbl, 4 ex
FIELD: food industry.
SUBSTANCE: invention relates to biotechnology, agriculture and zootechnics and may be used for industrial-scale breeding of broiler chickens of highly productive crosses. Prolyser symbiotic preparation based on Escherichia coli VL-613 is activated by way of resuspension with cooled boiled water or sterile physical solution at a temperature of 25°C - 37°C. The preparation in a dissolved form is given to drink to broiler chicken during 1 hour after the preparation activation in accordance with the following scheme. From the 8th day since the chicken birth and to the 22nd day the preparation is given in an amount of 100 mln microbial cells per day (per 1 chicken of "Кобб-500" and "Авиан-48" crosses) or 50 mln microbial cells per day (per 1 chicken of "Смена-7" cross). From the 22nd day and up to broiler chickens growing completion the preparation is given in an amount of 75 mln microbial cells per day (per 1 chicken of "Кобб-500" and "Авиан-48" crosses) or 50 mln microbial cells per day (per 1 chicken of "Смена-7" cross). The preparation is administered orally, preferably - during the morning feeding. The concentration of Escherichia coli VL-613 is equal to 1.75-3.05 billion microbial cells of the preparation per 1 broiler chicken throughout the whole growing cycle.
EFFECT: invention allows to fully replace synthetic lisyn in liquid given to drink to broilers, to increase poultry average daily weight gain, to increase the yield of Category 1 meat and to reduce the concentration of Escherichia coli VL-613 cells.
SUBSTANCE: method includes extraction of DNA from strains B.pertussis, performance of a polymerase chain reaction with simultaneous introduction of specific primers PF-PR for production of amplicons of a fragment of a promotor of pertussis toxin ptxP, restriction of produced amplicons with endonucleases Kpnl and Mlul and electrophoresis of restriction products in agarose gel. Differentiation of strains B.pertussis is carried out by comparison of size of amplicons of DNA fragments of promotor ptxP of investigated strains B.pertussis with reference samples of DNA strains B.pertussis.
EFFECT: invention may be used for accelerated detection of epidemic strains with modified genetic structure during microbiological and molecular-genetic monitoring of Bordetella pertussis strains and for further development of promising diagnostic and prevention preparations on their basis.
SUBSTANCE: strain is deposited under the number RCAM 00876 in the collection of All-Russia Research Institute of Agricultural Microbiology of the Russian Agricultural Academy. The strain Bacillus subtilis 8A has high fungicide activity against phytopathogenic activity against phytopathogenic fungi and bactericide activity against phytopathogenic bacteria, and also growth-stimulating activity relative to various crops.
EFFECT: strain may find application in agriculture as an efficient agent to increase productivity of plants and their protection against phytopathogenic microorganisms.
SUBSTANCE: protease is presented which has enhanced milk clotting activity, containing amino acid sequence at least 80 % identical to SEQ ID NO: 3, where the said protease has at least one mutation selected from the group consisting of: (a) substitution of glutamine, corresponding to glutamine at a position of 265 in SEQ ID NO: 3, with amino acid; and (b) replacement of glutamine, corresponding to glutamine at a position of 266 in SEQ ID NO: 3, with amino acid. DNA is described which encodes the said protease, the expression vector containing the said DNA, and the cell transformed with the said vector, designed for expression of the said protease. The method of production of protease having enhanced milk clotting activity is proposed, comprising culturing the said transformed cell in the cultural medium and isolation of protease from the cultural medium.
EFFECT: invention enables to obtain the protease with enhanced milk clotting activity.
16 cl, 2 dwg, 4 tbl
SUBSTANCE: preparation of thrombolytic and fibrinolytic action is produced by submerged cultivation on a liquid nutrient medium, of a strain of basidium fungus of Coprinus lagopides TI-12, or Coprinus lagopides TI-32, or Coprinus lagopides TI-33 stored in the high basidium fungi culture collection of the microbiological synthesis technology department of the St.-Petersburg State Technical University, separation of a mycelial biomass from a native solution. The prepared solution is concentrated by ultrafiltration with using a semipermeable membrane with a nominal molecular-mass rejection limit up to 20 kDa followed by cool dehumidification in the mode suitable for protein compounds. The invention has allowed detecting Coprinus lagopides from basidium fungi of Coprinus exhibiting high thrombolytic and fibrinolytic activity.
EFFECT: preparation in the low concentration shows a high level of thrombolytic and fibrinolytic activity in vivo.
2 dwg, 4 tbl, 5 ex
FIELD: food industry.
SUBSTANCE: oyster mushroom fruit bodies are homogenised, the obtained homogenate is frozen followed by defrosting and dividing into extract and residue. Residue is mixed with distilled water in mass ratio of 1:2, held at +4-+8°C, the secondary extract is separated and mixed with previously obtained extract. Food acid is added into the obtained mixture to achieve pH 3.8 and it is precipitated by saturation of the solution with sodium chloride at +4-+8°C. The obtained residue is separated and dried by freezing at -18°C. The dried residue is kept in a refrigerator. Before using, it is dissolved in a minimum amount of water and centrifuged.
EFFECT: high milk-clotting activity and increased ratio of milk-clotting and general proteolytic activity.
1 tbl, 2 ex
SUBSTANCE: method of obtaining milk-turning ferment includes deep cultivation of agaric macromycetes Coprinus lagopides on liquid medium, which contains sources of carbon, nitrogen and mineral salts.
EFFECT: ferment with high level of milk-turning activity and low level of general proteolytic activity.
FIELD: chemistry, biochemistry.
SUBSTANCE: strain of basidium fungus Trametes hirsuta (Wulfen) Pilat, deposited with collections of the OOO PKF "BIGOR" enumerated as CF-28, is extracted by repeated passages from monoglobular isolates of deep culture of the strain TsNIIMOD 56. The said strain features an active synthesis of extracellular laccase. The laccase activity makes some 20 to 26 ME/ml.
EFFECT: production of the basidium fungus strain with active extracellular laccase synthesis.
2 tbl, 2 ex
FIELD: chemistry, biochemistry.
SUBSTANCE: proposed method includes deep cultivation of the strain of fungi Trametes hirsuta (Wulfen) Pilat, OOO PKF "BIGOR" CF-28 on a nutrient medium, separation of the fungi biomass, membrane concentration of the filtrate. The nutrient medium composition incorporates growth factors in concentration of 0.1-1.02 wt % and wheat flour in concentration of 2.0-4.0 wt % is used as a source of carbon. The cultivation is carried out at 28 to 34°C, pH 4.0 to 4.5, the concentration of dissolved oxygen making at least 1.5 to 2.0 mg·dm-3. The membrane concentration is effected with the membrane element pore sizes varying from 0.1 to 0.5 microns, at a pressure difference of 1.0 to 4.0 MPa and temperature of 20 to 40°C. Laccase activity makes some 140 to 200 ME.
EFFECT: production of fermented laccase preparation with high activity.
FIELD: medicine; pharmacology.
SUBSTANCE: substance for dermatological medical products on the basis of a collagenase of a microbic parentage represents an ultrafiltrate allocated from a cultural liquid of Streptomyces lavendulae strain VKPM S-910 with collagenolytic activity of 1800-2500 KEA/ml and proteolytic activity of 120-200 PE/ml.
EFFECT: obtaining of biologically active ultrafiltrate with high collagenolytic and proteolytic activity for an effective utilisation as a part of various wound-healthing preparations and dermatology.
SUBSTANCE: disclosed is isolated polypeptide being acid-proof metalloprotease isolated from Thermoascus aurantiacus. Described are strain Thermoascus aurantiacus CGMCC No 0670 for polypeptide production and method for polypeptide production using said strain. Disclosed is method for plant protein treatment to increase digestion value thereof by using said polypeptide.
EFFECT: protease of good acid resistance useful in feed production.
6 cl, 7 ex, 4 tbl
SUBSTANCE: method provides the hydrolysis of starch-containing raw materials, following adding of the mineral salts to the obtained hydrolysate and medium fermentation with acid-forming fungi Aspergillus niger. The hydrolysis is carried out with ferment preparate of bacterial α-amilase at increased temperature and excessive pressure. The starch-containing raw material are rice flour allowed beforehand in the water at 20-25°C during 10-12 hrs. in amount of 205.3-235.7 g and rye flour in amount of 232.0-290.6 g. The carbon and nitrogen ratio in the obtained rice flour suspension is provided respectively equal (20.0-23.0):1, in rye flour suspension - (14.0-16.0):1. The ferment preparate is added up to achieving of carbon source dextrose equivalent value 35-40%. The sugars conversion to citric acid is 86.7-93.2%. α-amilase and glucoamylase activity is respectively 4.5-6.2 units A,C/cm3 and 160-180 units Gl.C/cm3.
EFFECT: increase of α-amilase and glucoamylase yield while maintaining the high yield of citric acid.
1 tbl, 7 ex