RU2202606C2 - Method for preparing vegetable raw to microbiological conversion - Google Patents

Abstract

FIELD: microbiological industry. SUBSTANCE: method involves preparing polysaccharide raw to microbiological conversion by its milling and homogenization in the dispersing device in the presence of derivatives of polyphosphoric acid soluble in nutrient medium of the general formula HO-[PO₃X]_n-PO₃X₂ where X is alkaline metal, hydrogen atom, ammonium ion or aliphatic radical and n ≥ 1. In preferable variant rotary-pulsating device of S-emulsifier type is used as dispersing device. Method provides decrease of treatment temperature of polysaccharide raw, decreased consumption of liquefying and saccharifying enzymes, reduction of process time of microbiological conversion, increased yield of end products, improved consumers' quality of end products. Invention can be used in production of enzymes, protein, ethyl alcohol, fodders and so on. EFFECT: improved method of preparing. 4 tbl, 18 ex

Description

 The invention relates to the microbiological industry, specifically to methods for preparing nutrient media based on polysaccharide raw materials, and can be used in the production of amylolytic enzymes, microbial protein, ethyl alcohol, antibiotics, vitamins, feed and other products of microbiological synthesis.

 Known methods for preparing a polysaccharide feedstock for microbiological conversion include the steps of grinding, homogenizing, and heat treating the feedstock at high temperature, followed by transferring it to culture media by processing polysaccharides (starch, cellulose, hemicellulose), protein, and other biopolymers and oligomers extracted from the feedstock. both with our own enzymes and exogenously introduced enzyme preparations in order to transfer them into nutrients assimilated by microorganisms.

A known method of producing microbial biomass, which consists in the cultivation of yeast of the genus Candida on a nutrient medium containing the main source of nutrition (n-paraffins) and an additional carbon source, which is a hydrolyzate of polysaccharide raw materials (wheat or rye bran) [Russian Federation Patent 2042713, cl. 6 C 12 P 21/00, C 12 N 1/16, 1/26 dated 03/31/94, B.I. 24 from 08.27.95]. According to the known method, an additional power source is obtained by hydrolysis of a polysaccharide feed in the presence of its own enzymes of the feed under conditions of elevated temperature (70-75 ^o C).

 The disadvantages of this method is the low yield of the finished product with a low crude protein content and an increased paraffin consumption coefficient, which is associated with the low quality of the nutrient medium due to the shallow dissolution of the polysaccharide raw materials (rye bran) assimilated by the yeast in the liquid phase.

A known method of preparing polysaccharide raw materials for microbiological conversion [USSR Author's Certificate 1081209, class. C 12 N 9/34, C 12 N 1/20], namely, a method of preparing a nutrient medium for the cultivation of glucoamylase producers, comprising preparing a suspension of flour in a mixer with a mixer (100-200 rpm), introducing amylolytic and cellulolytic enzymes into the suspension , fermentolysis of a suspension, addition of additional sources of nitrogen and mineral salts to the fermentalizate, followed by sterilization of the nutrient medium. In this case, the flour suspension is digested at 127-138 ^o С (pressure 1.5-2.5 atm) for 1.5-2 hours, followed by fermentolysis of the boiled flour with amylolytic and pectolytic or cellulolytic enzymes. Hydrolysis with enzymes is carried out at 80-95 ^o C for 1 h followed by sterilization of the medium. The medium prepared in this way is seeded with a glucoamylase producer, which is used as Aspergillus awamori, and the producer is cultured to the maximum glucoamylase activity in the medium for 110-120 hours.

A known method for the production of ethanol using a polysaccharide feedstock, including the stage of preparing the polysaccharide feedstock for microbiological conversion (fermentation) [Regulation for the production of alcohol from starchy raw materials. Part 1. Ministry of Food Industry. M .: VNIIIPrB, 1979, 270 s]. This method includes the steps of preparing raw materials for fermentation, fermentation and subsequent distillation and distillation of ethanol. The stage of preparation of raw materials for fermentation is that the raw materials are mechanically crushed, mixed in a chunk of a batch equipped with a mechanical stirrer, with water at a temperature of 45-55 ^o C and boiled at high temperatures of 135-145 ^o C and higher in cooking stations of various designs . The resulting mash (boiled flour suspension) is saccharified by enzymes (total consumption of α-amylase - 2.0-2.1 units of AC / g of conditional starch; consumption of glucoamylase - 6.0-6.2 units of HCl / g of conditional starch), conduct yeast generation and fermentation of saccharified wort, followed by isolation from a mature mash of alcohol and its rectification.

 The disadvantages of the described methods include: boiling polysaccharide raw materials at high temperatures, which requires the availability of special equipment operating at elevated pressure and the consumption of a significant amount of energy. In addition, as a result of activation of thermal degradation processes of flour nutrients, primarily starch and free sugars, significant losses of sugars, amino acids, vitamins and other growth factors utilized by the producer are observed, which reduces the yield of target products or requires the introduction of an increased amount of hydrolytic (amylolytic and glucolytic ) enzymes at the stage of fermentolysis of a boiled suspension of polysaccharide raw materials. Moreover, the maximum yield of the target product from a unit of raw material remains low. In addition, the nutrient medium obtained by the known method in the case of glucoamylase production is characterized by increased viscosity, which complicates the aeration of the medium, and therefore the cultivation process increases, and the glucoamylase activity in the medium decreases, which is also a disadvantage of the known method.

Closest to the proposed one is a method of preparing a polysaccharide feedstock for microbiological conversion, which involves grinding the feedstock, mixing the crushed feedstock with water (in a mixer equipped with a stirrer with a rotation speed of 100-200 rpm), its subsequent thermal exposure at a temperature of 90-105 ^o C and above with the further transfer of raw materials to the nutrient medium by processing it with enzyme preparations [Typical technological regulations for the production of alcohol from starchy raw materials. Ministry of Agriculture and Food of the Russian Federation. M., 1998, 79 s]. Specifically, the method is implemented in the production of ethyl alcohol and includes grinding starch-containing raw materials, preparing an aqueous suspension and homogenizing it in a batch chunk equipped with a stirrer, hydroenzymatic processing of the suspension, namely, preliminary heating of the suspension at a temperature of 50-55 ^o C with further exposure of the suspension in the hydrodynamic processing mode at a temperature from 75 ^o C to 95 ^o C, followed by aging the suspension at a temperature of 105-110 ^o C and a higher cooling and receiving the nutrient medium (wort) by saccharifying I received congestion enzymatic preparations, cooling and fermentation of saccharified wort bragorektifikatcii mature mash.

In accordance with the known method, in preparing starch-containing raw materials, for example rye, for fermentation, the processing temperature of the starch-containing suspension is 90-105 ^{° C.} , while the yield of ethyl alcohol does not exceed 65.5 dal of crude crude with 1 t of conventional starch of raw materials.

 The disadvantages of this method include the low yield of the target product, which, for example, in the case of ethanol production does not exceed 65.5 dal per ton of conventional rye starch, the high temperature of heat treatment of raw materials, which significantly reduces the quality of the wort due to inactivation of diluent enzymes and losses raw materials due to the occurrence of side (e.g., sugar-amine) reactions, high consumption of enzymes and high viscosity of nutrient media, especially when processing rye, which makes transportation difficult my suspension for technological communications. In addition, in real conditions of alcohol production by the known method, increased regulatory costs of amylolytic enzymes are practically observed (total consumption of α-amylase 2.5-3.1 units of AC / g of conditional starch; consumption of glucoamylase - 6.4-6.9 units (GlS / g of conditional starch and above), the losses of which are caused by their rapid inactivation during the heat treatment of polysaccharide raw materials at temperatures exceeding the optimum temperature of their action.

 The aim of the invention is to increase the yield of target products, reduce the temperature of heat treatment of polysaccharide raw materials, reduce the consumption of thinning and saccharifying enzymes, reduce the duration of the microbiological conversion process and improve the consumer qualities of the target products.

This goal is achieved by the fact that in the method of preparing the polysaccharide feed for microbiological conversion, including grinding and homogenization of the polysaccharide feed with its subsequent transfer to nutrient media, grinding and homogenization is carried out in a dispersing device in the presence of soluble in nutrient media derivatives of polyphosphoric acids of the general formula HO- [ PO ₃ X] _n —PO ₃ X ₂ , where X is an alkali metal, hydrogen, an ammonium ion or an aliphatic organic radical, and n≥1.

As dispersing devices can be used rotary pulsation apparatus, disintegrators, apparatus with high-speed mixers. The processing of suspensions of polysaccharide raw materials in dispersing apparatuses should ensure the flow of processes of grinding, homogenization and extraction of biopolymers from polysaccharide raw materials into the liquid (aqueous) phase. To carry out these processes, a rotor-pulsation apparatus or homogenizer can be used, in which the polysaccharide feedstock is processed in the gap between the stator and the rotating rotor at velocity gradients from 1.0 • 10 ⁵ m / s • m to 5.0 • 10 ⁵ m / s • m, apparatus with high-speed mixers in which the homogenization of raw materials is carried out at a speed of not less than 50 m / s. In this case, the proposed method is most effectively implemented in the case of the use of dispersing devices [International application PCT / RU / 96/00297, WO 98/16304 of 04.23.98, B 01 F 7/00], which work not only mechanically, but simultaneously and acoustic (mechano-acoustic) processing of the polysaccharide feed in the presence of soluble polyphosphoric acid derivatives of the above formula.

 So, for example, when preparing a nutrient medium for microbiological conversion, in the case of implementing the technology for producing ethanol in accordance with the proposed method, an active and deep dissolution of grain substances occurs, the intensification of liquefaction and saccharification processes, which are caused by a three-level effect on the grain structure.

 The first level of exposure (supramolecular level) is the deep mechanical grinding of grain grains and the mechano-acoustic homogenization of starch grains with the effective extraction of starch from them and its dissolution. This level is provided by the dispersion mode.

 Simultaneously with the first level, the second level of exposure (molecular level) is accompanied by processes of destruction and hydration of starch molecules (amylose and amylopectin), hemicelluloses and protein derivatives of polyphosphoric acids of the above formula, which intensifies the processes of dissolution of grain substances. Polyphosphoric acid derivatives, manifesting themselves mainly as complexing and / or hydrating substances, form with ions of two or more valence metals, as well as with biopolymer molecules (starch, non-starchy grain carbohydrates, proteins, including enzymes, lipids, etc. .) complex of variable composition complex highly hydrated compounds and, thus, contribute to the effective dissolution of grain substances, which leads, in the end result, to obtain such a technical result as an increase in ethanol yield, the reduction of the effective temperature of boiling polysaccharide raw materials, reducing the consumption of thinning and saccharifying enzymes. The intensification of the processes of liquefaction and saccharification by enzyme preparations is also due to the ability of polyphosphoric acid derivatives to reduce the viscosity of water-grain suspensions, as well as activate both their own amylolytic enzymes of grain and exogenously introduced in the implementation of the technology of alcohol production.

 The third level of exposure (at the level of a yeast cell) is the stimulation of alcoholic fermentation due to the effect of polyphosphoric acid derivatives of the above formula on cell energy. A yeast cell spends less wort sugars for its own energy needs, which is accompanied by additional alcohol production by the cell while reducing the duration of the fermentation process. In addition, lowering the boiling point of the polysaccharide feedstock, in the final result, not only leads to an increase in the yield of alcohol per unit of feedstock, but also favorably affects the improvement of the organoleptic characteristics of alcohol (prevention of the formation of microimpurities in a mature mash - substances of multistage reactions of thermal decomposition of grain substances and their bioconversion )

 In the case of, for example, the implementation of a technology for the cultivation of glucoamylase producers, which includes a method for preparing polysaccharide raw materials for microbiological conversion and providing for grinding and homogenizing polysaccharide raw materials in a dispersing device under chemomechanical processing conditions in the presence of soluble polyphosphoric acid derivatives soluble in nutrient media, the above formula is highly effective; raw materials and extraction of flour substances (including the whole enzyme comp Lex flour - amylases, proteinases, pectinases, etc.) into the liquid phase. Polyphosphoric acid-soluble derivatives in suspension, contributing to the dissolution (hydration) of starch in the liquid phase, reduce the viscosity of the suspension during its hydrolysis. In this case, the tertiary structure of starch molecules and proteins is destroyed without noticeable accumulation of monosugars in the nutrient medium (it is known that the presence of monosugars in the nutrient medium significantly inhibits the accumulation of glucoamylase).

Example 1 (According to the method of the Patent of the Russian Federation 2042713, CL 6 C 12 P 21/00, C 12 N 1/16, 1/26 from 03/31/94, B.I. 24 from 08/27/95)
Candida tropicalis VSB-928K yeast is grown continuously in a nutrient medium containing salts, g / l:
(NH ₄ ) ₂ SO ₄ - 1.5
H ₃ PO ₄ (70%) - 0.9
KCl - 0.4
MgSO ₄ • 7H ₂ O - 0.2
FеSO ₄ • 7Н ₂ О - 0.12
ZnSO ₄ • 7H ₂ O - 0.063
MnSO ₄ • 5H ₂ O - 0.033
The cultivation is carried out by feeding paraffin and polysaccharide raw materials (rye bran) in the form of a clarified hydrolyzate in the ratio (wt.%) Parahydrolyzate 1: 25 at a temperature of 32-34 ^o C, pH 4.0-4.2. The hydrolyzate is prepared by treating a 10-15 wt.% Aqueous suspension of bran at a temperature of 70-75 ^o With a pH of 5.0-6.0, with stirring, for 2.0-2.5 hours. The resulting hydrolyzate is clarified by centrifugation with the selection of the clarified part 60-70 vol.% and served in the growing process, and the concentrate formed during clarification of the hydrolyzate is combined with a stream of biomass suspension before drying in an amount of 10% of the total volumetric flow rate of the concentrate.

 Autohydrolyzed rye bran had the following indicators.

 1. The content of dry dissolved substances (SV,%) - 3.2.

 2. The content of reducing substances (PB,%) - 0.88.

 3. The flow time of the medium in an amount of 200 ml through an opening with a diameter of 4 mm is 14 s.

 The product yield from a given paraffin was 140%, the crude protein content of 53%. The consumption ratio of paraffin is 0.71 g / l. The consumption of paraffin is 3.2 wt.%. The results are presented in table 1.

Example 2 (the proposed method)
The continuous cultivation process is carried out analogously to example 1, with the only difference being that during the preparation of the hydrolyzate of polysaccharide raw materials (rye bran), the suspension is dispersed and homogenized by a S-emulsifier rotary pulsation apparatus [International application PCT / RU / 96/00297, WO 98 / 16304 dated 04/23/98, B 01 F 7/00] in the presence of a nutrient-soluble polyphosphoric acid derivative of the general formula HO- [PO ₃ X] _t -PO ₃ X ₂ , where X = Na, n = 1 from the calculation of 0, 0005-0.003 g per 1 g of conventional starch. Moreover, the processing of polysaccharide raw materials by S-emulsifier is carried out at two points in the process: at the stage of preparing an aqueous suspension of bran and before clarification of the hydrolyzate in centrifuges, but in both cases with a multiplicity of passage of the suspension of 1.0-1.3 through its working area. Next, the hydrolysis (autohydrolysis) of the bran suspension is carried out at a temperature of 58-65 ^o C, pH 5.0-6.0, with stirring, for 7.5-8.5 hours

 Autohydrolyzed rye bran had the following indicators.

 1. The content of dry dissolved substances (SV,%) - 8.1.

 2. The content of reducing substances (PB,%) - 3.28.

 3. The flow time of the medium in an amount of 200 ml through an opening with a diameter of 4 mm is 6 s.

 The product yield from a given paraffin was 171%, the crude protein content of 56%. The consumption ratio of paraffin is 0.57 g / l. Paraffin consumption of 2.4 wt.%. The results are presented in table 1.

Example 3
The process of continuous cultivation is carried out analogously to example 2, with the only difference being that the preparation of the hydrolyzate of polysaccharide raw materials (rye bran) excludes the use of the dispersion and homogenization method of S-emulsifier.

 Autohydrolyzed rye bran had the following indicators.

 1. The content of dry dissolved substances (SV,%) - 4.4.

 2. The content of reducing substances (PB,%) - 1.31.

 3. The flow time of the medium in an amount of 200 ml through an opening with a diameter of 4 mm is 8 s.

 The product yield from a given paraffin was 163%, the crude protein content of 54%. The paraffin consumption coefficient is 0.66 g / l. The consumption of paraffin is 3.0 wt.%. The results are presented in table 1.

Example 4
The process of continuous cultivation is carried out analogously to example 2, with the only difference being that in the preparation of the hydrolyzate of polysaccharide raw materials (rye bran), the use of a soluble polyphosphoric acid derivative of the indicated formula in the nutrient medium is excluded.

 Autohydrolyzed rye bran had the following indicators.

 1. The content of dry dissolved substances (SV,%) - 4.9.

 2. The content of reducing substances (PB,%) - 1.3.

 3. The flow time of the medium in an amount of 200 ml through an opening with a diameter of 4 mm is 11 s.

 The yield of the product from a given paraffin, as well as the content of crude protein, the consumption coefficient of paraffin and the consumption of paraffin were the same as in example 3. The results are presented in table 1.

 As can be seen from the data presented in table 1, the highest yield of the target product is observed in the case of processing the polysaccharide raw materials (suspension of rye bran) with a dispersant in the presence of polyphosphoric acid derivatives of the above formula, that is, by the proposed method.

Example 5
50% of the calculated amount of water is poured into a medium-pressure vessel equipped with a mechanical stirrer (100-200 rpm), the stirrer is turned on and, having heated the water to 45-55 ^o С, 40% of the calculated amount of wheat flour is added (the finished suspension should contain 14-22 wt.% solids), then the remaining amount of water is poured, the remaining amount of flour is added, the pH of the suspension is adjusted to 5.0 and 0.3 g of amylosubtiline G10x, 0.11 g of Celloviridin G3x and 4.8 g of Pectofoetidine are added G3x dissolved in water (0.6 L), then injected soluble in flour suspension produced the bottom of polyphosphoric acid of the general formula HO- [PO ₃ X] _n -PO ₃ X ₂ , where X = Na, n = 1 at the rate of 0.23 g per liter of suspension. Next, the suspension is subjected to dispersion and homogenization for 1 min in a remote dispersing device, for example, an S-emulsifier, in which the suspension is processed in the gap between the stator and the rotating rotor at a velocity gradient of 5.0 • 10 ⁵ m / s • m under exposure conditions on the medium being processed resonant vibrations of the rotor and stator, which are determined by the value of the maximum amplitude of sound vibrations. In the process of dispersion, the temperature of the suspension is raised to 50 ^o C, maintained at this temperature for 80 minutes, then the pH of the suspension is set to 6.0 and 0.3 g of amylosubtiline G10x dissolved in water is added, the temperature is raised to 70 ^o C, maintained at this temperature 20 minutes. The obtained hydrolyzate should not change color when stained with iodine. Then, ointment of malt sprouts, 0.75 L, (NH ₄ ) ₂ SO ₄ , 90 g, KH ₂ PO ₄ , 6.6 t, MgSO ₄ , 7.5 g, is introduced into the hydrolyzate obtained. The medium thus prepared is sterilized at 121 ^o C for 40 min, cooled, seeded with Aspergillus awamori glucoamylase producer and cultured at 33 ^o C for 120 hours with aeration and stirring.

 The biochemical composition of the nutrient medium had the following indicators.

 1. The content of monosugars in terms of glucose,% - 1.2.

 2. The content of alcohol-soluble dextrins,% - 5.0.

 3. The content of alcohol-insoluble dextrins,% - 3.9.

 4. The content of undissolved starch,% - 0.12.

 5. The flow time of the medium in an amount of 500 ml through an opening with a diameter of 4 mm is 9 s.

 Samples are taken during cultivation. The activity of glucoamylase in deep culture after 96 and 120 hours of cultivation, respectively, is 133 and 142 units / ml (according to the glucose oxidase method).

In the control experiment during the cultivation of the same strain on a medium prepared by a known method [USSR Author's Certificate 1081209, class. C 12 N 9/34, C 12 N 1/20], the activity of glucoamylase in a deep culture at 120 h of growth is 121 units / ml. Moreover, in the known method, the cooking of flour suspension is carried out at a temperature of 127-138 ^o With, that is, 58-69 ^o With more than in the proposed method.

 The biochemical composition of the nutrient medium had the following indicators.

 1. The content of monosugars in terms of glucose,% - 0.9.

 2. The content of alcohol-soluble dextrins,% - 2.4.

 3. The content of alcohol-insoluble dextrins,% - 6.3.

 4. The content of undissolved starch,% - 2.8.

 5. The flow time of the medium in an amount of 500 ml through an opening with a diameter of 4 mm is 17 s.

Example 6
Polysaccharide raw materials (rye) are crushed on a roller and / or hammer mill to a grinding quality of at least 70-90% (passage of crushed grain through a sieve with a hole diameter of 1 mm) and mixed with warm reflux water or warm condensation water of the rectification unit in a pre-weld mixer horizontal type in a ratio of 1: 2.5-3.5, respectively. If necessary, the batch in the mixer is heated to 65-70 ^o With hot and / or dead steam and kept at this temperature for at least 45-55 minutes At the same time as crushed grain and water are fed into the mixer, a suspension of α-amylolytic (diluting) enzymes is fed from the corresponding collectors at a rate of 60-70% of the total number of diluting enzymes and an aqueous suspension of polyphosphoric acid-derived derivatives of the general formula HO- [PO ₃ X] _n -PO ₃ X ₂ , where X = Na, n = 1 from the calculation of 0.0005-0.003 g per 1 g of conventional starch. Next, the prepared batch is subjected to fine grinding and homogenization using a dispersing unit in the gap between the stator and the rotating rotor at a velocity gradient of 5.0 • 10 ⁵ m / s • m, for example, an S-emulsifier in the mode of at least one passage through the installation working zone . The homogenized and partially hydrolyzed kneading with the same dispersing unit is fed into the apparatus (apparatuses) of the chemomechanical-enzymatic processing of kneading (KhMFO), in which the kneading is subjected to heat treatment at a temperature of 68-72 ^o C.

The housing of the KhMFO apparatus is made vertical, cylindrical, with a conical bottom; the apparatus is equipped with one circulation circuit with centrifugal pumps (one of them is reserve). The strapping and construction of the internal overflow pipes of the KhMFO-1 and KhMFO-2 devices should provide the maximum possible path for the passage of the water-grain mass during the KhMPO-processing. Next, the mass from the KhMFI apparatus (s) at a temperature of 68-72 ^o C is fed into a combined vacuum cooler and saccharifier, where the remaining amount of the fluidizing enzyme is introduced, i.e. 40-30% of the total amount of fluidizing enzyme, and a sufficient amount of saccharifying enzyme. The saccharified wort is then fed to a heat exchanger for cooling, where it is cooled to 22-32 ^° C, fermented using thermo-tolerant yeast races, followed by isolation and rectification of the alcohol using a distillation unit.

 The results of production experiments on the influence of the modes (modes 4, 9, 14, 19) of preparing the polysaccharide feed for fermentation and technobiochemical parameters of the wort (nutrient media based on the polysaccharide feed) are presented in Tables 2 and 4, respectively.

As can be seen from the data in Table 2, it is not possible to realize the alcohol production process at a temperature of 68-72 ^° C (mode 19) without using the polyphosphoric acid derivative of the above formula and S-emulsifier.

 Separate use of the polyphosphoric acid derivative of the above formula (regimen 9, and the S-emulsifier (regimen 4) slightly improves the technical and economic indicators of alcohol production (alcohol yield per ton of conventional starch, its organoleptic characteristics, reduction in the consumption of enzyme preparations), but does not lead to significant improvement of the above indicators.

 The best results were obtained when the process was carried out in mode 14 (according to the proposed method): the alcohol yield was 66.8 dal per ton of conventional starch, and the total enzyme consumption was lower than in all tested modes, and amounted to 1 for α-amylase, 8 units AC / g conditional starch, for glucoamylase - 5.5 units. GLS / g conditional starch.

Example 7
The method is carried out analogously to example 6, however, a polyphosphoric acid derivative of the formula HO [PO ₃ X] _n -PO ₃ X ₂ , where X = K, n = 2, is used, the dispersion and homogenization of the batch is carried out in the dispersing apparatus of the disintegrator in the gap between the stator and the rotating rotor with a velocity gradient value of 1.0 • 10 ⁵ m / s • m. Processing of the batch coming from the stage of its preparation is carried out in three stages. At the first stage - in the first KhMPO-apparatus, the batch is kept at a temperature of 68-70 ^o С, at the second stage - in the second KhMPO-apparatus - at a temperature of 90-98 ^o С, and at the third stage - in a tubular sterilizer and steam separator-storage - at a temperature of 100-105 ^o C. Heating of the mass in the tubular sterilizer is carried out with hot steam using a contact head mounted on the mass supply line to the tubular sterilizer.

 The results of experiments on the influence of the mode (modes 3, 8, 13, 18) of preparing the polysaccharide raw materials for fermentation on the indicators of alcohol production and technical and biochemical parameters of the wort are presented in Tables 2 and 4, respectively.

As can be seen from the data in table 2, as the temperature of cooking raw materials decreases in known methods [the Regulation for the production of alcohol from starchy raw materials. Part 1. Ministry of Food Industry. M .: VNIIIPrB, 1979, 270 s; Typical technological regulations for the production of alcohol from starchy raw materials. The Ministry of Agriculture and Food of the Russian Federation, M., 1998, 79 S. ] from 135-145 ^o С (mode 16) to 110-120 ^o С (mode 17) and, further, up to 90-105 ^o С (mode 18), the amount of alcohol output per ton of conventional starch has a steady tendency to decrease, and consumption of α-amylase and glucoamylase, on the contrary, tends to increase. A further decrease in the cooking temperature exacerbates the picture and makes the cooking process technologically unjustified.

However, the cooking process at a temperature of 90-105 ^o With possible and economically justified only if the organization of the process of preparing the polysaccharide raw materials for microbiological conversion mode 13, i.e. according to the proposed method, under the conditions of dispersion and homogenization of the batch in the presence of derivatives of polyphosphoric acids of the above formula. In this case, in comparison with mode 16 and mode 18, the alcohol yield increases from 65.3 dal / t of conventional starch to 66.3 dl / t of conventional starch in mode 13, while the consumption of enzymes decreases from 2.0 units . AC / g conditional starch and 6.2 units. GLS / g conditional starch in the regime of 16 to 1.7 units. AC / g conditional starch and 5.0 units GLS / g of conditional starch in mode 13. Separate use of technological methods of dispersion (mode 3) and treatment with derivatives of polyphosphoric acids (mode 8) somewhat improves the technical and economic parameters of the process compared to mode 18, but does not reach the indicators obtained in mode 13, those. by the proposed method.

Example 8
The method is carried out analogously to example 6, only the preparation of the batch and the boiling of the water-grain mass is carried out at a temperature of 58-62 ^o C, i.e. at that temperature when it is technologically feasible and possible to have spatio-temporal combination of such technological operations as preparation of a batch and its KhMFO processing with simultaneous dilution and saccharification of the cooked masses.

When implementing this method, the preparation of the batch is carried out at a temperature of 58-62 ^o C, and the calculated amount of the polyphosphoric acid derivative of the general formula HO [PO ₃ X] _n -PO ₃ X ₂ , where X = NH ₄ , n = 5, the total estimated amount of a fluidizing enzyme (α-amylase) and the estimated amount of a saccharifying enzyme (glucoamylase). The saccharification stage is excluded from the scheme, and the boiled and saccharified mass from the KhMPO apparatus (s) is fed directly to the heat exchanger for cooling, followed by fermentation. With the exclusion of capacitive apparatus from the scheme, saccharification of mash requires compensation of these volumes due to an equivalent increase in the volume of the KhMFO apparatus (s).

 The experimental results on the influence of the temperature regime of raw material digestion (modes 5, 10, 15, 20) on the technical and economic indicators of alcohol production and the technical and biochemical parameters of wort are presented in Tables 2 and 4, respectively.

 As can be seen from the data in Table 2, it seems possible to implement the process of alcohol production, which is attractive from the point of view of hardware design, when the processes of raw material boiling, liquefaction, and saccharification are combined in time and space. An indispensable condition for the effective implementation of this process is the fine grinding of grain in a dispersing device with the simultaneous chemical treatment of derivatives of polyphosphoric acids in the presence of water. Under these conditions (mode 15), the highest alcohol yield was obtained among all the presented options for cooking raw materials (modes 1-20), which amounted to 67.9 dal / ton of conventional starch. Separate use of the method of dispersing the batch (mode 5) or processing it with derivatives of polyphosphoric acids (mode 10), and even more so their absence (mode 20) does not give the desired results. This is especially indicative of the example of the implementation of regime 20, when the alcohol yield turned out to be very low, and the enzyme consumption was the highest, while the duration of the fermentation process increased sharply.

Example 9
According to the technology of example 6, the process of producing alcohol is carried out, however, as a polyphosphoric acid derivative, a compound of the general formula HO [PO ₃ X] _n -PO ₃ X ₂ , where X = H, n = 6, is used, while at the stage of preparing the batch in water cereal suspension is injected with an aqueous solution of sodium carbonate (soda) or ammonia.

 The technical and economic efficiency of the process is similar to the process in mode 13 (Tables 2 and 4).

Example 10
According to the technology of example 6, the process of producing alcohol is carried out, which includes the stage of preparing the polysaccharide raw material for microbiological conversion, however, a device with a high-speed stirrer is used as a dispersing device, in which dispersion and homogenization processes are carried out at a speed of 51.4 m / s, and as a polyphosphoric derivative acids are used compounds of the general formula HO- [PO ₃ X] _n -PO ₃ X _2, where X = ethyl, n = 2. In this case, the yield was 66.8 alcohol gave / t conditional starch, wherein the flow enzymes analogs ene flow regime 14 enzymes (Table 2 and 4).

Example 11
According to the technology of example 6, the process of producing alcohol is carried out, however, as a polyphosphoric acid derivative, a mixture (wt.% 1: 1) of compounds of the general formula HO- [PO ₃ X] _n -PO ₃ X ₂ , where X = ethyl, n = 1, is used and X = K, n = 1. The technical and economic efficiency of the process is similar to the process in mode 14 (Tables 2 and 4).

Example 12
Upon receipt of the alcohol, the method of preparing the polysaccharide raw materials for fermentation is carried out analogously to example 6, however, the digestion of the water-grain batch is carried out not in the KhMPO apparatus (s), as in the case of example 1, but in the cooking station in the continuous brewing mode [Regulation for the production of alcohol from starchy raw materials. Part 1. Ministry of Food Industry. M .: VNIIIPrB, 1979, 270 s] at a temperature of 135-145 ^o C. According to this method, simultaneously with the supply of crushed grain and water to the mixer, a suspension of α-amylolytic (diluting) enzymes is fed at the rate of 10-15% of the total amount of diluting enzymes and an aqueous solution of soluble in a nutrient medium derivatives of polyphosphoric acids of the general formula HO- [PO ₃ X] _n -PO ₃ X ₂ , where X = K, n = 2. During the preparation of the batch, the water-grain suspension is treated in an apparatus equipped with a high-speed mixer in which mixing and homogenization is carried out with awn 50-58 m / s.

 The remaining amount of α-amylase (85-90% of the calculated total amount) and the estimated amount of glucoamylolytic enzyme are set at the saccharification stage.

 The results of experiments on the influence of the mode (modes 1, 6, 11, 16) of preparing starchy raw materials for fermentation on the technical and economic indicators of alcohol production are presented in Tables 2 and 4.

As can be seen from the data in Tables 2 and 4, the processing of the batch with a dispersant (high-speed mixer) and / or soluble derivatives of polyphosphoric acids of the above formula with subsequent boiling of the batch in the temperature range 135-145 ^o С positively affected the technical and economic indicators alcohol production. However, this positive effect for various modes of preparing starchy raw materials for fermentation is diverse in nature, which is due to the technological limits of the possibility of the operation of the cooking station with varying mutually affecting process parameters - the effective temperature of the kneading of the batch and its degree of homogeneity.

 When implementing the process of alcohol production in the case of processing the batch only with a high-speed mixer (mode 1), in comparison with the known method (mode 16), it is possible to reduce the consumption of α-amylase by 0.2 units without deteriorating the quality of the wort. AC / g conditional starch and glucoamylase consumption by 0.3 units. GLS / g conditional starch. In addition, the duration of fermentation is reduced from 72 hours to 62 hours, i.e. for 10 hours

However, an increase in the yield of alcohol per ton of conventional starch (mode 1) compared with the known method (mode 16) is not observed, which is associated with the inevitable digestion of a well-homogenized batch (noticeable darkening of the cooked mass) at established high boiling points (135-145 ^o С )

 A similar picture can be observed by comparing the cooking processes of raw materials in modes 6 and 16: in this case, only the use of derivatives of polyphosphoric acids leads to savings in effective doses of thinning and saccharifying enzymes, to a reduction in the duration of fermentation, but also due to an increase in the effect of weight transfer, increase no alcohol output is observed.

 Combining processing modes (mode 11), i.e. using the method of dispersing the batch in the presence of derivatives of polyphosphoric acids, it is possible to significantly reduce the consumption of thinning and saccharifying enzymes to 0.9 units. AC / g conditional starch and 4.4 units. GLS / g conditional starch, respectively.

Thus, the separate use of a dispersant or derivatives of polyphosphoric acids in the technology of kneading when implementing a high-temperature mode of boiling (135-145 ^o С) is possible and desirable only with the aim of reducing the consumption of enzyme preparations for liquefaction and saccharification and in order to reduce the duration of fermentation.

Example 13
The method implements analogously to example 6, only the cooking temperature is maintained at 110-120 ^o C (modes 2, 7, 12, 17).

 The results of experiments on the influence of the temperature regime (modes 2, 7, 12, 17) of preparing the polysaccharide feed for fermentation on the technical and economic indicators of alcohol production are presented in Tables 2 and 4.

 As can be seen from the data in Tables 2 and 4, a decrease in the temperature of raw material digestion by a known method (mode 17) is not economically feasible. When implementing this method, the cooking temperature of the raw materials is insufficient, which leads to incomplete dissolution of grain starch in a nutrient medium and, as a result, to an increase in the level of undissolved starch in a mature mash, an increase in the content of the residual amount of unfermented dry substances in the mash (the value of "brood") and an increase in the amount of unfermented sugars compared to the process in mode 16. The process of alcohol production in mode 17 is characterized by a decrease in the yield of alcohol to 65.3 dal / ton of conventional starch against the background of an increase in the consumption of thinning enzymes (up to 2.5 units of AS / g of conditional starch) and saccharifying enzymes (up to 6.4 units of HCl / g of conditional starch).

Processing of the batch with either a dispersant (high-speed mixer) or polyphosphoric acid derivatives (modes 2 and 7) with further boiling of the batch at a temperature of 110-120 ^o C improves the technical and economic indicators of alcohol production (the alcohol yield increases to 65.7 dal / ton of conventional starch and 65.9 dal / ton of conventional starch), respectively. At the same time, in regimes 2 and 7, the consumption of both thinning and saccharifying enzymes decreases compared to regime 17 while reducing the duration of the fermentation process.

 Better results were obtained by homogenizing the batch with a dispersant (high-speed stirrer) in the presence of polyphosphoric acid derivatives, i.e. during the implementation of regime 12. In this case, the alcohol yield was 66.0 dal / ton of conventional starch, the consumption of the fluidizing enzyme was reduced to 1.1 units. AC / g conditional starch, saccharification enzyme consumption - up to 4.5 units. HlS / g conditional starch, and the duration of fermentation is reduced by 10 hours compared with the known method (mode 16) (table 2).

Example 14
According to the technology of example 6, the process of producing alcohol is carried out, however, as the polyphosphoric acid derivative, a calcium compound insoluble in a batch of the general formula HO- [PO ₃ X] _n -PO ₃ X ₂ , where X = Ca, n = 15, is used. The technical and economic efficiency of the process is similar to the process according to mode 4 (processing of the batch with a dispersant only). Thus, polyphosphoric acid derivatives insoluble in the batch do not have any positive effect on the process of alcohol production.

Example 15 (prototype method)
Polysaccharide raw materials (rye) crushed by hammer crushers (the passage of crushed grain through a sieve with a hole diameter of 1 mm at least 75-80%) is mixed with warm water and a bacterial enzyme preparation - a source of α-amylase (amylosubtilin Gx or G3x), maintaining the kneading temperature at the level of 50-55 ^o C. Amylosubtilin is dosed at the rate of 1.5-2.0 units. AC / g conditional starch. The ratio of grain and water entering the mixer is 1: 3.0-3.2 and is set depending on the starchiness of the grain, taking into account that the concentration of wort in the saccharifier is 16-18% by saccharimeter. The duration of the kneading in the mixer is 10-12 minutes. Then the batch, heated with the help of a contact head to a temperature of 70-72 ^o C, is kept for 2.0-2.5 hours in the apparatus of hydrodynamic and enzymatic processing of the first stage (GDFO-1) with constant mixing of the batch. To better maintain the activity of α-amylase, its introduction is carried out in 2 points: 1.5 units AC / g conditional starch in the mixer and 0.5 units AC / g conditional starch in the apparatus GDFO-1. Next, the batch is fed into the apparatus of the hydrodynamic and enzymatic treatment of the second stage (GDFO-2), where the mass passing through the sections of the apparatus is heated from 65 ^o C to 85 ^o C with subsequent exposure of the mass at a temperature of 105-110 ^o C.

 To saccharify a sterilized and chilled mass, it is mixed with amylosubtilin G3x at the rate of 0.4-0.6 units. AC / g conditional starch and glucavamorin Gx at the rate of 6.2-6.4 units. GLS / g conditional starch. Next, the obtained sugared wort is cooled and fermented, and rectified alcohol is isolated from the mature mash.

 The results of production experiments are presented in table 2 (mode 17) and 4. As can be seen from the data in table 2, when implementing the process according to the prototype method, the yield of alcohol per ton of conditional starch was 65.3 gave with a fermentation duration of 76 hours

Example 16 (by the method of the Rules for the production of alcohol from starchy raw materials. Part 1. The Ministry of food industry. M: VNIIIPrB, 1979, 270 S.)
The polysaccharide raw materials (rye) crushed by hammer crushers (the passage of crushed grain through a sieve with a hole diameter of 1 mm at least 50-60%) is mixed with warm water, maintaining the batch temperature at 65-70 ^o C. The ratio of grain to water entering the mixer-pre-welder is 1: 2.5-3.5 and is set depending on the starchiness of the grain, taking into account that the concentration of wort in the saccharifier is 16-18% by saccharimeter. The duration of the kneading in the mixer is 10-12 minutes. The cooked batch is fed to the heater, where it is heated with secondary steam to 75-85 ^o C and then fed to the contact head, where the batch is heated to a temperature of 135-145 ^o C, followed by its supply to the cooking station.

 To saccharify a sterilized and chilled mass, it is mixed with amylosubtilin G3x at the rate of 1.9-2.1 units. AC / g conditional starch and glucavamorin Gx at the rate of 6.0-6.4 units. GLS / g conditional starch. Next, the obtained sugared wort is cooled and fermented, and rectified alcohol is isolated from the mature mash.

 The results of production experiments are presented in table 2 (mode 16). As can be seen from the data of table 2, when implementing the process according to the known method, the yield of alcohol per ton of conventional starch was 65.5 gave with a fermentation duration of 72 hours

Example 17
According to the technology of example 6, the process of producing alcohol is carried out, however, orthophosphoric acid is used as a catalyst for the hydrolysis of the polysaccharide feed at a rate of 0.003 g per 1 g of conventional starch of raw materials, while the batch is processed in the S-emulsifier in the gap between the stator and the mouth at a speed gradient 5.0 • 10 ⁵ m / s • m.

 As a result of a production experiment, the yield of the target product with 1 ton of conventional starch was 65.3 dal, which almost corresponds to the yield of alcohol during the process of preparing starch-containing raw materials for fermentation using only a dispersing device (Table 2, mode 4).

Example 18
According to the technology of example 6, the process of producing alcohol is carried out, however, sodium orthophosphate in the amount of 0.003 g per 1 g of conventional starch of raw materials is used as a catalyst for the hydrolysis of the polysaccharide feedstock, while the batch is processed in the dispersing device in the same way as in example 17.

 As a result of the experiment, the alcohol yield was 65.1 yields per ton of conventional starch of raw materials, which is lower than the alcohol yield during the preparation of the polysaccharide raw material for fermentation using a dispersing device (see Table 2, mode 4).

 In the obtained mash (distillation of the direct distillation of the mature mash) in all realized modes of preparing the polysaccharide raw material for microbiological convection in the technology for producing ethanol (modes 1-20), the analysis of the content of microimpurities in the mash was carried out by chromatography-mass spectrometry. In addition, tasting of rectified alcohol obtained in the processes according to modes 1-20 was carried out (the conditions of rectification, as well as the method and apparatus and technological design of the distillation unit were the same in all cases). The data are presented in table.3.

 As can be seen from the data table. 3, the maximum amount of microimpurities in the distillate distillates was found in the case of preparing the polysaccharide feed for microbiological conversion, where the cooking temperature of the feed was maximum (mode 6), while provoking the process of digesting the mass by dispersing and / or introducing derivatives of polyphosphoric acids (modes 1.6 and , especially, 11) contributed to an increase in trace elements in mature mash. Lowering the temperature of heat treatment of raw materials in the preparation of polysaccharide raw materials for microbiological conversion in accordance with the proposed method leads to a significant reduction in trace elements in mature mash.

 The qualitative and quantitative composition of trace elements affects the tasting (organoleptic) characteristics of rectified alcohol: with a decrease in their quantity and variety, organoleptic indicators of alcohol improve. So, the alcohols obtained in modes 4, 5, 9, 10, 14, 15, 19, 20, compared with the alcohols obtained in the processes under modes 1, 2, 3, 6, 7, 8, 11, 12, 13 , 16, 17, 18, had a pronounced, without extraneous smack, but moderate alcohol smell and taste. These alcohols were characterized by the absence of a pungent odor, a sweetish taste, without a taste of bitterness, and greatly weakened by burning of the oral mucosa.

Claims (1)

A method of preparing a polysaccharide feedstock for microbiological conversion, including grinding and homogenizing the polysaccharide feedstock with its subsequent transfer to nutrient media, characterized in that the grinding and homogenization is carried out in a dispersing device in the presence of polyphosphoric acid derivatives of the general formula HO- [PO ₃ X soluble in nutrient media] ] _n —PO ₃ X ₂ , where X is an alkali metal, hydrogen, an ammonium ion or an aliphatic organic radical, and n≥1.

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