L-alanine, preparation method and fermentation device

Abstract

The invention relates to L-alanine, a preparation method and a fermentation device, and relates to the technical field of L-alanine. The culture medium contains a carbon source, a nitrogen source, a phosphorus source and trace elements; the L-alanine is synthesized by the microorganisms by controlling fermentation conditions and time; the invention further relates to an L-alanine preparation method, an L-alanine preparation device is adopted, the L-alanine preparation device comprises a fermentation tank and a stirrer for mixing a plurality of raw materials of L-alanine, and the stirrer is further provided with a treatment part for treating floating foam, oily substances and impurities generated in the stirring process. The processing part can collect floating foam on the surface of the mixed solution and can also collect impurities and oily substances in the mixed solution, and meanwhile an opening and closing door on the processing part can be opened and closed, so that the impurities difficult to capture in the mixed solution fluctuation state are collected in a unified mode; and the removal rate of floating foam and impurities is greatly improved.

Description

Technical Field

[0001] The present invention relates to the technical field of L-alanine, and in particular to L-alanine, a preparation method and a fermentation device. Background Art

[0002] Alanine is an important amino acid, which is widely used in medicine, food additives and daily chemical products. There are several main methods for its preparation: chemical synthesis, enzyme catalysis, microbial fermentation and hydrolysis extraction.

[0003] Among them, microbial fermentation is currently a more common industrial production method, with the advantages of low cost and environmental protection. The fermentation method uses microorganisms such as Pseudomonas to ferment and synthesize the target product, with a short fermentation cycle and high conversion rate.

[0004] For example, Chinese patent publication number CN116144471A discloses a novel proline fermentation device and fermentation process thereof, comprising: a fermentation tank and a foam cleaning mechanism; by setting up the foam cleaning mechanism, the foam cleaning mechanism can use the air introduced into the fermentation tank to blow the foam on the top of the fermentation liquid to achieve the purpose of cleaning the foam, compared with a rake-type defoamer, the defoaming effect is improved, compared with a scraper-type defoamer and a disc-type defoamer, the energy consumption is reduced and the vibration of the fermentation tank is reduced, and compared with chemical defoaming, the chance of contamination is reduced.

[0005] However, the above-mentioned proline preparation device still has some shortcomings in actual use: 1. First, in the above-mentioned prior art, the bubbles floating on the top of the fermentation liquid can be eliminated by the foam cleaning mechanism, and its main principle is to actively puncture the bubbles by blowing. However, it should be noted that it has an excellent treatment effect mainly for some larger bubbles, and the generation of large bubbles is often accompanied by a large number of small bubbles, and its effect on the treatment of small bubbles is not good.

[0006] 2. Secondly, in the prior art, during the fermentation period of the product, not only bubbles are generated, but also some substances can be observed floating on the surface of the fermentation liquid. These substances may be bacteria, proteins or polysaccharides produced during the growth of microorganisms, or they may be oils or other organic substances produced during the fermentation process. The prior art cannot clean these substances, which will affect the efficiency of the fermentation process or the purity of the product.

[0007] Therefore, under the above-stated viewpoint, the existing L-alanine production equipment still has room for improvement. Summary of the invention

[0008] In order to solve the above problems, the present invention provides an L-alanine, a preparation method and a fermentation device, which adopt the following technical solutions: In a first aspect, the present invention relates to L-alanine, which is prepared by fermenting Escherichia coli with a culture medium containing a carbon source, a nitrogen source, a phosphorus source and trace elements: Escherichia coli is used as the host strain, and the culture medium containing carbon source, nitrogen source, phosphorus source and trace elements is as follows: Carbon source: Glucose accounts for 15-36g / L; Nitrogen source: Amino acids account for 6-12 g / L; Phosphorus source: Phosphate accounts for 0.1-0.5 g / L; Trace elements: Iron accounts for 0.1-1.5mg / L; zinc accounts for 0.1-1.5mg / L; magnesium accounts for 0.5-11mg / L; The balance is pure water.

[0009] In a second aspect, the present application also relates to a method for preparing L-alanine, and the method for preparing L-alanine is as follows: S1: pre-culturing the Escherichia coli bacterial liquid containing Escherichia coli, and then preparing the culture medium, and preparing various raw materials required for providing nutrition and fermentation for Escherichia coli, that is, mixing the carbon source, nitrogen source, phosphorus source and trace elements to prepare the culture liquid; S2: preparing a static fermentation tank, mixing the Escherichia coli liquid and the culture liquid and adding them into the fermentation tank for culture and fermentation; monitoring the cell density and alanine production parameters in the fermentation tank; S3: A stirrer is provided in the fermentation tank, and a processing component is provided in the stirrer. The stirrer in the fermentation tank is started to uniformly stir the Escherichia coli liquid and the culture solution in the fermentation tank through the stirrer to make them fully mixed, and then the impurities floating in the mixed solution and the foam generated during the stirring process are processed by the processing component; S4: After the fermentation is completed, alanine is extracted from the fermentation broth by an extraction method, and centrifugation and filtration are used for preliminary extraction; S5: purifying the extracted alanine by removing impurities using chromatography technology to obtain a high-purity alanine product; S6: Conduct quality inspection on the purified alanine, including purity and content indicators, to ensure that the product meets the requirements, and then package the alanine products that meet the quality requirements.

[0010] In the third aspect, the present application discloses an L-alanine fermentation device, which includes a fermentation tank and a stirrer, wherein the stirrer includes a stirring motor, a driving gear, a stirring column and a stirring elbow, the stirring motor is installed on the fermentation tank through a bracket, the output end of the stirring motor faces downward and is connected to a control gear, the stirring column is penetrated into the interior of the stirring column along the height direction of the fermentation tank, and a control gear is also installed on the top of the stirring column, the control gear on the stirring column is meshed with the control gear on the stirring motor, and the stirring elbow is arranged on the stirring column at equal intervals along the length direction of the stirring column.

[0011] Preferably, the processing component comprises a lifting frame and a processing frame integrally connected to the lifting frame, two groups of collecting sponges are slidably arranged in the processing frame, each group of collecting sponges is slidably arranged in the processing frame along the length direction of the processing frame, the lifting frame is arranged on the stirring column, and the processing frame is installed on the outer side wall of the lifting frame; A feed trough and a discharge trough for collecting and removing the froth generated in the process of preparing the L-alanine solution are respectively provided on both sides of the processing frame.

[0012] Preferably, two groups of symmetrically distributed opening and closing doors are hinged on the feed chute and the discharge chute of the processing frame, and the two groups of opening and closing doors cover the feed chute and the discharge chute.

[0013] Driving gears for driving the doors to open and close are provided on the axes of the two groups of opening and closing doors. Several of the driving gears are rotatably fitted on the side walls of the processing frame. Driving racks are meshed on the driving gears. The driving racks are slidably distributed along the height direction of the processing frame. A limiting frame for limiting the movement of the driving racks is also provided on the side walls of the processing frame close to the driving gears, and a reset tension spring is connected between the driving racks and the limiting frame.

[0014] Preferably, sawtooth structures are alternately provided on both sides of the driving rack, and the two driving racks are respectively meshed with a plurality of driving gears on both sides of the processing frame.

[0015] Preferably, each group of the collecting sponges also includes a plurality of cleaning cotton pieces for cleaning floating foam, and linkage mounting plates are snap-fitted onto the side walls of the plurality of cleaning cotton pieces, and the linkage mounting plates on the two groups of cleaning cotton pieces in the processing frame are staggered.

[0016] Preferably, a linkage seesaw is hingedly installed on one side of the processing frame, and one side of the linkage seesaw is slidably engaged with the linkage mounting plate on one side of the processing frame, and the other side of the linkage seesaw is engaged with the linkage mounting plate on the other side of the processing frame, and the other side of the linkage seesaw is engaged with the linkage mounting plate on the other side of the processing frame.

[0017] Preferably, the fermentation tank is also provided with a linkage component, the linkage component includes two pressing knobs, the two pressing knobs are slidably arranged on the fermentation tank, and a No. 1 rod is installed at the bottom of one side of the pressing knob, and a start ring is provided at one end of the No. 1 rod away from the pressing knob. A second rod is installed at the bottom of the pressing knob on the other side. A cleaning ring is arranged at one end of the second rod away from the pressing knob. The bottom of the cleaning ring abuts against a cleaning rod, which is arranged at one side of the linkage seesaw.

[0018] Preferably, flushing pipes are provided on both sides of the processing frame in the width direction. The flushing pipe is inserted into the stirring column on the side away from the processing frame and extends outward along the stirring column to the outside of the fermentation tank. The flushing pipe is connected to an external water source on the side away from the processing frame.

[0019] Preferably, the starting ring and the cleaning ring are slidably arranged inside the fermentation tank along the height direction thereof.

[0020] In summary, the present application includes at least one of the following beneficial technical effects: 1. The processing component in the present application can not only collect the foam on the surface of the mixed solution, but also collect the impurities and oily substances in the mixed solution. At the same time, the opening and closing door on the processing component can be opened and closed, so as to collect all the impurities that are difficult to capture in the fluctuating state of the mixed solution, thereby greatly improving the removal rate of foam and impurities.

[0021] 2. The collecting sponge in the present application can be switched alternately to achieve filtering and cleaning of the collecting sponge, which greatly improves the efficiency of the mixed solution in removing impurities and foam. At the same time, the collecting sponge is cleaned in time after collecting impurities, which can effectively increase its service life.

[0022] Third, the vibration component of the present invention can vibrate the cleaning cotton piece in the collection sponge, and actively puncture the floating foam through the collection sponge, which can greatly improve the rapid crushing of small-volume floating foam, thereby greatly improving the efficiency of floating foam removal. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0024] Figure 1 It is a structural schematic diagram of the main body of the present invention.

[0025] Figure 2 It is a cross-sectional view of a fermentation tank of the present invention.

[0026] Figure 3 It is a schematic diagram of the structure of the processing component of the present invention from a first perspective.

[0027] Figure 4It is a schematic diagram of the structure between the processing frame and the processing components of the present invention.

[0028] Figure 5 The present invention Figure 4 A partial enlarged view of point A in the figure.

[0029] Figure 6 It is a schematic diagram of the structure of the processing component of the present invention from a second viewing angle.

[0030] Figure 7 It is a third perspective structural schematic diagram of the processing component of the present invention.

[0031] Figure 8 It is a structural schematic diagram of the linkage component of the present invention.

[0032] Fig. 9 It is a schematic structural diagram of the vibration component of the present invention from a first viewing angle.

[0033] Fig.10 The present invention Fig. 9 A partial enlarged view of point B in FIG.

[0034] Fig.11 It is a schematic structural diagram of the vibration component of the present invention from a second viewing angle.

[0035] Fig.12 It is a flow chart of the method for preparing L-alanine of the present invention.

[0036] Explanation of the reference numerals: 1. fermentation tank; 2. agitator; 3. processing component; 20. stirring motor; 21. control gear; 22. stirring column; 23. stirring elbow; 30. lifting frame; 31. processing frame; 32. collecting sponge; 33. feeding trough; 34. discharging trough; 310. opening and closing door; 311. driving gear; 312. driving rack; 313. limiting frame; 314. reset spring; 35. cleaning cotton piece; 36. linkage mounting plate; 37. linkage seesaw; 4. linkage component; 40. pressing knob; 41. rod one; 42. starting ring; 43. rod two; 44. cleaning ring; 45. cleaning rod; 46. flushing pipe; 5. vibration component; 50. vibration ring; 51. vertical rod; 52. vertical spring; 53. vibration rod; 54. vibration spring. DETAILED DESCRIPTION

[0037] The following is combined with Figure 1-Figure 12 This application is described in further detail.

[0038] The examples of the present application disclose L-alanine and a preparation method thereof.

[0039] Firstly, it involves L-alanine, which is prepared by fermenting Escherichia coli with a culture medium containing a carbon source, a nitrogen source, a phosphorus source and trace elements.

[0040] L-alanine is produced by fermentation of the following raw materials, namely, Escherichia coli bacterial solution and culture medium containing carbon source, nitrogen source, phosphorus source and trace elements; by controlling the fermentation conditions and time, microorganisms can synthesize L-alanine.

[0041] The formula of the culture medium is as follows: carbon source: glucose accounts for 15-36g / L; nitrogen source: amino acid accounts for 6-12g / L; phosphorus source: phosphate accounts for 0.1-0.5g / L; trace elements: iron accounts for 0.1-1.5mg / L; zinc accounts for 0.1-1.5mg / L; magnesium accounts for 0.5-11mg / L; the balance is pure water.

[0042] The fermentation method of L-alanine is mainly achieved through the microbial fermentation process. This method has the advantages of environmental protection and low cost, and is one of the more mainstream processes in industrial production. The key to the production of L-alanine by fermentation is to select the appropriate strain and optimize the fermentation conditions.

[0043] Embodiment 1: Among them, the inoculation amount of Escherichia coli is 10^7CFU / mL.

[0044] Fermentation medium: Carbon source: glucose 20g / L; Nitrogen source: amino acids account for 8g / L; Phosphorus source: phosphate accounts for 0.1g / L; Trace elements: iron accounts for 1.2mg / L; zinc accounts for 1mg / L; magnesium accounts for 0.4mg / L; The balance is pure water.

[0045] Operation steps: pre-culture the fermentation medium, add 200g of glucose, 80g of amino acids, 1g of phosphate, 12mg of trace element iron, 10mg of trace element zinc and 4mg of trace element magnesium into a 10L fermentation tank 1, and mix with the aid of pure water.

[0046] Then, the temperature of the fermentation tank 1 was heated to 32 degrees Celsius for constant temperature, and then 100 mL of Escherichia coli was inoculated into the fermentation tank 1. The stirrer 2 was controlled at a speed of 300 rpm to uniformly stir the mixed solution in the fermentation tank 1, and the stirring fermentation was carried out for 36 hours.

[0047] Then, stirring of the mixed solution in the fermentation tank 1 was stopped and the mixture was allowed to ferment for 8 hours. Thus, a mixed solution containing L-alanine was obtained, wherein the synthesis efficiency of L-alanine reached a maximum of 348 g per hour when the synthesis rate was stable. Finally, the content of L-alanine in the mixed solution was measured to be 8256 g.

[0048] The product was then extracted by centrifugation, filtration and purification, and finally 7954 g of L-alanine was obtained.

[0049] By preparing L-alanine through this method, the conversion rate of L-alanine preparation can be effectively improved and the loss of L-alanine can be reduced.

[0050] Embodiment 2: The inoculum size of E. coli was 10^7 CFU / mL.

[0051] Fermentation medium: Carbon source: glucose 30g / L; Nitrogen source: amino acids account for 11g / L; Phosphorus source: phosphate accounts for 0.1g / L; Trace elements: iron accounts for 1.0mg / L; zinc accounts for 0.5mg / L; magnesium accounts for 0.4mg / L; The balance is pure water.

[0052] Operation steps: pre-culture the fermentation medium, add 300g of glucose, 110g of amino acids, 1g of phosphate, 10mg of trace element iron, 5mg of trace element zinc and 4mg of trace element magnesium into a 10L fermentation tank 1, and mix with the aid of pure water.

[0053] Then, the temperature of the fermentation tank 1 was heated to 32 degrees Celsius for constant temperature, and then 100 mL of Escherichia coli was inoculated into the fermentation tank 1. The stirrer 2 was controlled at a speed of 300 rpm to uniformly stir the mixed solution in the fermentation tank 1, and the stirring fermentation was carried out for 36 hours.

[0054] Then, stirring of the mixed solution in the fermentation tank 1 was stopped and the mixture was allowed to ferment for 8 hours. Thus, a mixed solution containing L-alanine was obtained, wherein the synthesis efficiency of L-alanine reached a maximum of 398 g per hour when the synthesis rate was stable. Finally, the content of L-alanine in the whole mixed solution was measured to be 9346 g.

[0055] The product was then extracted by centrifugation, filtration and purification, and finally 8933 g of L-alanine was obtained.

[0056] Embodiment three: Among them, the inoculation amount of Escherichia coli is 10^7CFU / mL.

[0057] Fermentation medium: Carbon source: glucose 36g / L; Nitrogen source: amino acids account for 12g / L; Phosphorus source: phosphate accounts for 0.5g / L; Trace elements: iron accounts for 1.2mg / L; zinc accounts for 1mg / L; magnesium accounts for 0.4mg / L; The balance is pure water.

[0058] Operation steps: pre-culture the fermentation medium, add 360g of glucose, 120g of amino acids, 5g of phosphate, 12mg of trace element iron, 10mg of trace element zinc and 4mg of trace element magnesium into a 10L fermentation tank 1, and mix with the aid of pure water.

[0059] Then, the temperature of the fermentation tank 1 was heated to 32 degrees Celsius for constant temperature, and then 100 mL of Escherichia coli was inoculated into the fermentation tank 1. The stirrer 2 was controlled at a speed of 300 rpm to uniformly stir the mixed solution in the fermentation tank 1, and the stirring fermentation was carried out for 36 hours.

[0060] Then, stirring of the mixed solution in the fermentation tank 1 was stopped and the mixture was allowed to ferment for 8 hours. Thus, a mixed solution containing L-alanine was obtained, wherein the synthesis efficiency of L-alanine reached a maximum of 437 g per hour when the synthesis rate was stable, and finally the content of L-alanine in the whole mixed solution was 9620 g.

[0061] The product was then extracted by centrifugation, filtration and purification, and finally 9312 g of L-alanine was obtained.

[0062] It can be concluded that carbon source is an important energy source for bacterial growth and metabolism. An appropriate amount of carbon source can promote bacterial growth and the synthesis of metabolites, thereby increasing the production of L-alanine. Nitrogen source is an important raw material for protein synthesis. Different contents of nitrogen source will directly affect the synthesis efficiency of L-alanine. As the nitrogen source content is continuously added within a reasonable range, the synthesis efficiency of L-alanine can be effectively improved.

[0063] According to the formula in the present application, the efficiency of L-alanine synthesis and conversion can be effectively improved, the cost consumption can be greatly reduced, and the preparation output of L-alanine can also be increased.

[0064] After metabolic engineering, Escherichia coli can efficiently produce L-alanine through fermentation under anaerobic conditions. This method not only increases the yield of L-alanine, but also reduces production costs, making this process significantly advantageous in both economic and environmental benefits.

[0065] In addition, the fermentation process of producing L-alanine is usually carried out at room temperature and pressure, using a "one-tank" fermentation method, which can achieve zero carbon dioxide emissions, further improving its environmental friendliness. This process not only reduces dependence on non-renewable petroleum-based products, but also significantly reduces production costs through technological innovation, allowing the application of L-alanine to be expanded to multiple emerging fields such as daily chemicals, medicines and health products, as shown below.

[0066] See also Fig.12 As shown, the present application also proposes a method for preparing L-alanine.

[0067] S1: Pre-culture the Escherichia coli bacterial liquid containing Escherichia coli, then prepare the culture medium, and prepare various raw materials required for providing nutrition and fermentation for Escherichia coli, that is, mix the carbon source, nitrogen source, phosphorus source and trace elements to prepare the culture liquid.

[0068] Recombinant E. coli strains were constructed by genetic engineering. For example, the L-alanine dehydrogenase gene was introduced into E. coli, and genes that may compete with the pyruvate metabolic pathway, such as the lactate dehydrogenase gene and the pyruvate formate lyase gene, were knocked out to ensure that pyruvate was preferentially converted into L-alanine.

[0069] S2: preparing a fermentation tank 1, mixing the Escherichia coli liquid and the culture liquid and adding them into the fermentation tank 1 for culture and fermentation; monitoring parameters such as the cell density and alanine production in the fermentation tank 1.

[0070] The required sugar source, nitrogen source and inorganic salt are put into the fermentation tank 1 at one time. After sterilization and cooling, the cultured seeds are moved in for fermentation. During the fermentation process, anaerobic fermentation can be used, with glucose as the raw material, and the target product L-alanine is synthesized by microbial fermentation through the transformation and regulation of the bacterial metabolic pathway. During the fermentation process, it is necessary to control the pH value and provide nitrogen and carbon sources, for example, by adding ammonia water and glucose to maintain the pH of the fermentation liquid.

[0071] S3: starting the agitator 2 in the fermentation tank 1, stirring the E. coli liquid and the culture solution in the fermentation tank 1 at a uniform speed through the agitator 2 to make them fully mixed, and then treating the impurities floating in the mixed solution and the foam generated during the stirring process through the processing component 3.

[0072] S4: After the fermentation is completed, alanine is extracted from the fermentation liquid by an extraction method, and a preliminary extraction is performed by centrifugation and filtration.

[0073] S5: further purifying the extracted alanine by using chromatography technology to remove impurities and obtain a high-purity alanine product.

[0074] S6: Conduct quality inspection on the purified alanine, including purity and content indicators, to ensure that the product meets the requirements, and package the alanine products that meet the quality requirements.

[0075] Further, with respect to the L-alanine preparation method, the present application also proposes an L-alanine preparation device.

[0076] Its application in the process of L-alanine preparation can technically remove excessive foam generated in the process of L-alanine preparation. If a large amount of foam floats on the surface of the L-alanine solution, the foam will cover the surface of the L-alanine solution, and severe hypoxia will occur during the fermentation process, thereby affecting the stability of L-alanine fermentation.

[0077] Especially when cleaning the floating foam; after the floating foam is cleaned, there will still be some oily substances and some other impurities floating on the surface of the L-alanine solution, and the processing component 3 in the present application can not only clean the floating foam but also remove other impurities floating on the surface of the L-alanine solution.

[0078] See also Figure 1 As shown, it is a schematic diagram of the structure of the fermentation tank 1 and the interior of the fermentation tank 1 in the present application; the L-alanine preparation device includes a stationary fermentation tank 1 and a stirrer 2 for mixing and stirring several raw materials of L-alanine inside the fermentation tank 1, and the stirrer 2 is also provided with a processing component 3 for processing foam, oily substances and impurities generated during the stirring process.

[0079] It should be noted that the main function of the stirrer 2 is to stir the L-alanine fermentation. The stirrer 2 can stir the various raw materials of L-alanine to improve its uniformity, thereby indirectly improving its fermentation efficiency.

[0080] First, the cover of the fermentation tank 1 is opened, and then the raw materials required for the preparation of L-alanine are poured into the fermentation tank 1, and then stirred by the stirrer 2, as shown below.

[0081] Reference Figure 2 As shown, that is, a schematic diagram of the structure of the agitator 2 in the present application; specifically, the agitator 2 includes a stirring motor 20, a stirring column 22 and a stirring elbow 23. The stirring motor 20 is installed on the fermentation tank 1 through a bracket, and the output end of the stirring motor 20 faces downward and is connected to the stirring column 22. The stirring column 22 is arranged inside the stirring column 22 along the height direction of the fermentation tank 1, and the stirring elbow 23 is arranged on the stirring column 22 at equal intervals along the length direction of the stirring column 22.

[0082] It should be noted that the stirring column 22 is disposed inside the fermentation tank 1 .

[0083] During the specific implementation process, the stirring motor 20 is started, and the stirring motor 20 controls the stirring column 22 to rotate at a uniform speed. The stirring column 22 controls the stirring elbow 23 to stir during the uniform rotation. The stirring elbow 23 and the stirring column 22 mix the mixed solution inside the fermentation tank 1 in this process to ensure the fermentation of various raw materials required for L-alanine after mixing.

[0084] The present application is mainly prepared by a fermentation method, which synthesizes L-alanine by microbial fermentation. This method has the advantages of environmental protection and low cost, and the fermentation process can be carried out in a large fermentation tank 1, and the production scale can be adjusted as needed. The fermentation method makes full use of ecological products, the production process is clean and efficient, and zero carbon dioxide emissions can be achieved.

[0085] When the stirring elbow 23 stirs the mixed solution, a large number of bubbles will be generated first. These bubbles will accumulate due to the long-term stirring operation, and it will be more difficult to remove these bubbles after accumulation.

[0086] In addition, during the stirring process, fermentation will be carried out simultaneously, so some biological components such as proteins and polysaccharides will be produced and float on the surface of the mixed solution; further, microorganisms will produce microbial metabolites during metabolism, including bacteria, pigments and colloidal substances, etc., and then some oily substances will appear when the mixed solution is stirred. If the oily substance cannot be dissolved in time, it will completely cover the surface of the mixed solution, causing necrosis of microorganisms that require oxygen in the solution, thereby affecting the fermentation preparation of L-alanine.

[0087] Therefore, in response to the above problems, the present application proposes a processing component 3 to solve the above problems, as shown below.

[0088] See also Figure 3 , Figure 4 , Figure 5 and Figure 6 As shown, it is a processing component 3 for cleaning floating objects and foams on the surface of the mixed solution in the fermentation tank 1 in the present application. The processing component 3 includes a lifting frame 30 and a processing frame 31 integrally connected to the lifting frame 30. Two groups of collecting sponges 32 are arranged in the processing frame 31, and each group of collecting sponges 32 is slidably arranged in the processing frame 31; the lifting frame 30 is arranged on the stirring column 22, and the processing frame 31 is installed on the outer wall of the lifting frame 30.

[0089] A feed trough 33 and a discharge trough 34 are respectively provided on both sides of the processing frame 31 for collecting and removing the froth generated during the preparation of the L-alanine solution.

[0090] It should be noted that the lifting frame 30 is composed of two C-shaped semicircular structures, and the lifting frame 30 is installed on the stirring column 22 by bolts and nuts. It should be noted that the lifting frame 30 can be lifted and lowered along the stirring column 22.

[0091] After the fermentation tank 1 is filled with a number of raw materials for preparing L-alanine, the stirring column 22 is stirred, and the processing frame 31 on the stirring column 22 rotates synchronously. During the rotation of the processing frame 31, the mixed solution in the fermentation tank 1 enters the interior of the processing frame 31 from the feed slot 33 of the processing frame 31, and then the mixed solution passes through the feed slot 33 of the processing frame 31, and then passes through the collecting sponge 32 and flows out from the discharge slot 34 of the processing frame 31; in this process, the collecting sponge 32 collects the foam and quickly eliminates the foam; further, the collecting sponge 32 adsorbs impurities in the mixed solution passing through the processing frame 31.

[0092] See also Figure 6 As shown, it is a schematic diagram of the structure of the collecting sponge 32 in the present application. Each group of collecting sponges 32 also includes a plurality of cleaning cotton pieces 35 for cleaning floating foam. Linkage mounting plates 36 are snap-fitted onto the side walls of the plurality of cleaning cotton pieces 35, and the linkage mounting plates 36 on the two groups of cleaning cotton pieces 35 in the processing frame 31 are staggered.

[0093] It should be noted that, in the present application, there are two groups of collecting sponges 32, and the two groups of collecting sponges 32 are staggered up and down, and each group of collecting sponges 32 is provided with multiple groups of cleaning cotton pieces 35. The functions of the cleaning cotton pieces 35 of each group of collecting sponges 32 are different. For the convenience of description, the cleaning cotton pieces 35 in each group of collecting sponges 32 are arranged in order as cleaning cotton piece No. 1 35, cleaning cotton piece No. 2 35 and so on.

[0094] One of the cleaning cotton pieces 35 is close to the feed slot 33 of the processing frame 31, and its surface is uneven, and one end close to the feed slot 33 is a spike structure. When the processing frame 31 rotates at a constant speed, the floating foam collected inside it will contact the spike structure of the cleaning cotton piece 35, quickly puncturing the foam, thereby removing the foam.

[0095] The impurities and oily substances collected in the processing frame 31 are adsorbed and collected by the second cleaning cotton sheet 35 to prevent them from re-entering the mixed solution in the fermentation tank 1.

[0096] Furthermore, in order to ensure the efficiency and integrity of collecting foam and impurities, the present application also proposes an opening and closing door 310 with an adjustable angle.

[0097] When the stirring column 22 is stirring, the mixed solution in the fermentation tank 1 is in a fluctuating state, and the foam on the surface of the mixed solution fluctuates up and down synchronously with the mixed solution; similarly, impurities floating on the surface of the mixed solution will also float in the shallow area of ​​the mixed solution due to the fluctuation. In order to ensure the integrity of the foam and impurities collected by the processing frame 31 and avoid poor collection efficiency of the processing frame 31, the processing frame 31 can open the opening and closing door 310 when collecting it, as shown below.

[0098] Replay Figure 4 and Figure 5 As shown, two symmetrically distributed opening and closing doors 310 are provided between the feed trough 33 and the discharge trough 34 of the processing frame 31, and a driving gear 311 for driving the opening and closing of the opening and closing door 310 is provided on the axis thereof, and a driving rack 312 is meshed on the driving gear 311, and the driving rack 312 is slidably distributed along the height direction of the processing frame 31, and a limit frame 313 for limiting the movement of the driving rack 312 is also provided on the processing frame 31, and a reset tension spring 314 is connected between the driving rack 312 and the limit frame 313.

[0099] It should be noted that, in the initial state, the opening and closing doors 310 on both sides of the processing frame 31 are in a closed state, and the opening and closing doors 310 cover the feed trough 33 and the discharge trough 34 of the processing frame 31. When it is necessary to collect the foam and impurities floating on the surface of the mixed solution, the opening and closing doors 310 are opened, and the opening and closing doors 310 are tilted to ensure that the opening and closing doors 310 at the feed trough 33 position of the processing frame 31 can absorb a wider range of mixed solutions, ensuring that impurities on the surface of the mixed solution and in the shallow layer of the mixed solution can be collected by the processing frame 31. At the same time, after the impurities are collected inside the processing frame 31, the impurities are adsorbed onto the collecting sponge 32 under the inertia of the rotation of the processing frame 31. At this time, the foam quickly bursts after contacting the collecting sponge 32, thereby achieving the purpose of eliminating the foam.

[0100] After the foam on the surface of the mixed solution is removed, the impurities and oily substances floating on the surface of the mixed solution are also absorbed by the collecting sponge 32 .

[0101] After the opening and closing door 310 is unfolded, the opening and closing door 310 on one side penetrates into a certain height above the upper end of the mixed solution. When the processing frame 31 rotates, the opening and closing door 310 on one side can collect the floating foam accumulated on the surface of the mixed solution to prevent the floating foam from accumulating too high and being unable to be cleaned; further, the opening and closing door 310 on the other side is inserted into the lower end of the mixed solution and is located in the shallow area of ​​the mixed solution. When the processing frame 31 rotates, the opening and closing door 310 on the other side will collect impurities floating in the shallow area of ​​the mixed solution into the processing frame 31 until they are collected by the collecting sponge 32.

[0102] Therefore, the processing component 3 in the present application can not only collect the foam on the surface of the mixed solution, but also collect the impurities and oily substances in the mixed solution. At the same time, the opening and closing door 310 on the processing component 3 can be opened and closed, so as to collect all the impurities that are difficult to capture in the fluctuating state of the mixed solution, thereby greatly improving the removal rate of foam and impurities.

[0103] The collecting sponge 32 in the present application can be switched alternately to achieve simultaneous filtering and cleaning of the collecting sponge 32, which greatly improves the efficiency of the mixed solution in removing impurities and foam. At the same time, the collecting sponge 32 is cleaned in time after collecting impurities, which can effectively improve its service life.

[0104] Reference Figure 7 , Figure 8 and Fig. 9 As shown, in order to further ensure the efficiency and stability of cleaning the collecting sponge 32, the present application also proposes a linkage component 4, which can not only improve the convenience of operation, but also clean the collecting sponge 32, ensuring that the collecting sponge 32 can be quickly cleaned before being blocked, so that it can achieve the best performance in adsorbing impurities, and indirectly improve the service life of the collecting sponge 32.

[0105] Specifically, a linkage component 4 is also provided on the fermentation tank 1, and the linkage component 4 includes two push knobs 40, which are slidably arranged on the fermentation tank 1, and a No. 1 rod 41 is installed at the bottom of one side of the push knob 40, and a start ring 42 is provided at the end of the No. 1 rod 41 away from the push knob 40.

[0106] Sawtooth structures are alternately arranged on both sides of the driving rack 312 , and the driving rack 312 is arranged between the driving gears 311 of the processing frame 31 .

[0107] The starting ring 42 and the cleaning ring 44 are limited and can only move along the height direction of the fermentation tank 1 but cannot rotate.

[0108] A second rod 43 is installed at the bottom of the pressing knob 40 on the other side. A cleaning ring 44 is provided at one end of the second rod 43 away from the pressing knob 40 . A cleaning rod 45 abuts against the bottom of the cleaning ring 44 . The cleaning rod 45 is provided on one side of the linkage rocker 37 .

[0109] It should be noted that after pressing the push knob 40, the push knob 40 will move downward and will not rebound due to friction. Only after it is pulled upward can it return to the initial state.

[0110] During the specific implementation process, when the stirring motor 20 is started and the mixed solution in the fermentation tank 1 is stirred and fermented, the pressing knob 40 on one side is pressed first. At this time, the pressing knob 40 on one side controls the starting ring 42 to move downward through the No. 1 rod 41 until the starting ring 42 squeezes the driving rack 312, and the driving rack 312 moves downward after being squeezed. The driving rack 312 drives the driving gear 311 to rotate. After rotating, the driving gear 311 controls the two opening and closing doors 310 on the same side of the processing frame 31 to rotate relative to each other until the two opening and closing doors 310 on the same side of the processing frame 31 are opened.

[0111] When the opening and closing door 310 is opened, the processing frame 31 filters the mixed solution in the fermentation tank 1. As the fermentation tank 1 continues to ferment, the processing frame 31 also continues to remove the floating foam and impurities in the mixed solution. However, as the collecting sponge 32 in the processing frame 31 adsorbs impurities for a long time, its impurity adsorption performance will gradually decrease until it can no longer adsorb impurities. Therefore, in order to avoid the reduction and failure of its impurity adsorption performance, the present application also provides another pressing knob 40.

[0112] Pull the push knob 40 on one side upward to the initial state, and then press the push knob 40 on the other side downward. The push knob 40 on the other side controls the cleaning ring 44 to move downward through the No. 2 rod 43. At this time, the cleaning ring 44 controls the cleaning rod 45 to squeeze one side of the linkage seesaw 37. At this time, the cleaning cotton piece 35 located at the top and used in the processing frame 31 will move downward with the linkage seesaw 37, and the unused cleaning cotton piece 35 on the other side of the linkage seesaw 37 will move upward. The unused cleaning cotton piece 35 rests on the top of the processing frame 31, and the mixed solution in the fermentation tank 1 can continue to be cleaned of impurities. The used cleaning cotton piece 35 at the bottom of the processing frame 31 is simultaneously washed with water, and the used cleaning cotton piece 35 is high-pressure rinsed, and the adsorbed impurities will be flushed out in the reverse direction and processed uniformly.

[0113] A groove is provided between the linkage seesaw 37 and the processing frame 31 for the linkage seesaw 37 to rotate, but it should be noted that a rubber sleeve is provided here to divide the processing frame 31 into two relatively closed spaces, one side is used to filter the mixed solution, and the other side is used to collect the cleaning of the sponge 32.

[0114] Flushing pipes 46 are provided on both sides of the processing frame 31 in the width direction. The flushing pipe 46 is inserted into the stirring column 22 on the side away from the processing frame 31, and extends outward along the stirring column 22 to the outside of the fermentation tank 1. The flushing pipe 46 is rotatably connected to an external water source on the side away from the processing frame 31.

[0115] It should be noted that when the stirring column 22 rotates, the flushing pipe 46 also rotates therewith. However, it should be noted that when the flushing pipe 46 rotates, the end connected to the water source can cause it to rotate, but it does not affect the transportation of the water source. At the same time, the rotating position is sealed to prevent water leakage.

[0116] Look again Figure 7 As shown, specifically, a linkage seesaw 37 is hingedly installed on one side of the processing frame 31, one side of the linkage seesaw 37 is slidably engaged with a linkage mounting plate 36 on one side of the processing frame 31, and the other side of the linkage seesaw 37 is engaged with a linkage mounting plate 36 on the other side of the processing frame 31.

[0117] During the specific implementation process, a linkage seesaw 37 is provided in the processing frame 31, and one side of the linkage seesaw 37 is slidably connected to a group of collecting sponges 32 through a mounting plate, and the other side of the linkage seesaw 37 is slidably connected to another group of collecting sponges 32 through a linkage mounting plate 36, so that the two groups of collecting sponges 32 can be raised and lowered alternately; ensuring that the two groups of collecting sponges 32 can operate alternately, greatly improving the removal rate of impurities and floating foam collection.

[0118] See also Fig. 9 , Fig.10 and Fig.11 As shown, in order to further ensure the efficiency of foam removal, the present application also proposes a vibration component 5, which includes a vibration ring 50, a vertical rod 51, a vertical tension spring 52, a vibration rod 53, and a vibration tension spring 54. The vibration ring 50 is arranged on the top of the fermentation tank 1, and the vertical rod 51 is slidably arranged on the processing frame 31 along the height direction of the fermentation tank 1, and the top of the vertical rod 51 abuts against the vibration ring 50, a vertical tension spring 52 is arranged between the vertical rod 51 and the processing frame 31, and one end of the vertical rod 51 located in the processing frame 31 abuts against the vibration rod 53, and a vibration tension spring 54 is connected between the vibration rod 53 and the processing frame 31, and the end of the vertical rod 51 and the vibration rod 53 close to each other is a wavy structure.

[0119] The vibration ring 50 cannot rotate or move, and the vertical rod 51 rotates along the bottom of the vibration ring 50 following the processing frame 31 .

[0120] During the specific implementation process, when the processing frame 31 rotates at a uniform speed, the vertical rod 51 on the processing frame 31 moves back and forth along the vibration ring 50 driven by the vertical tension spring 52. During the reciprocating up and down movement of the vertical rod 51, the wave-like structure of the vertical rod 51 reciprocates and squeezes the vibration rod 53, and then the vibration rod 53 quickly vibrates the No. 1 cleaning cotton sheet 35 of each group of collection sponges 32, ensuring that the No. 1 cleaning cotton sheet 35 can quickly remove the floating foam in the mixed solution, thereby greatly improving the efficiency of floating foam cleaning.

[0121] During operation: In the first step, the cover of the fermentation tank 1 is opened, and then the raw materials required for preparing L-alanine are poured into the fermentation tank 1 .

[0122] Step 2: Start the stirring motor 20, which controls the stirring column 22 to rotate at a constant speed. The stirring column 22 controls the stirring elbow 23 to stir during the constant rotation. The stirring elbow 23 and the stirring column 22 mix the mixed solution inside the fermentation tank 1 in this process to ensure the fermentation of various raw materials required for L-alanine after mixing.

[0123] Step 3: When the stirring elbow 23 stirs the mixed solution, a large number of bubbles will be generated first. These bubbles will accumulate due to the long-term stirring operation, and it will be more difficult to remove these bubbles after accumulation; In addition, fermentation will be carried out simultaneously during the stirring process, so some biological components such as proteins and polysaccharides will be produced and float on the surface of the mixed solution.

[0124] Step 4: When the stirring column 22 is stirring, the processing frame 31 on the stirring column 22 rotates synchronously. During the rotation of the processing frame 31, the mixed solution in the fermentation tank 1 enters the interior of the processing frame 31 from the feed slot 33 of the processing frame 31, and then the mixed solution passes through the feed slot 33 of the processing frame 31, and then passes through the collecting sponge 32 and flows out from the discharge slot 34 of the processing frame 31; in this process, the collecting sponge 32 collects the foam and quickly eliminates the foam; further, the collecting sponge 32 adsorbs impurities in the mixed solution passing through the processing frame 31.

[0125] Step 5: When the stirring column 22 is stirring, the mixed solution in the fermentation tank 1 is in a fluctuating state. At this time, the foam on the surface of the mixed solution fluctuates up and down synchronously with the mixed solution; similarly, the impurities floating on the surface of the mixed solution will also float in the shallow area of ​​the mixed solution due to the fluctuation. In order to ensure the integrity of the foam and impurities collected by the processing frame 31 and avoid poor collection efficiency of the processing frame 31, the processing frame 31 can open the opening and closing door 310 when collecting it, so as to improve the efficiency of impurity collection.

[0126] Step 6: When the opening and closing door 310 is opened, the processing frame 31 filters the mixed solution in the fermentation tank 1. As the fermentation in the fermentation tank 1 continues, the processing frame 31 also continuously removes the floating foam and impurities in the mixed solution.

[0127] The embodiments of this specific implementation method are all preferred embodiments of the present invention, and are not intended to limit the protection scope of the present invention. Therefore, any equivalent changes made based on the structure, shape, and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. L-alanine is prepared by fermenting Escherichia coli with a culture medium containing a carbon source, a nitrogen source, a phosphorus source and trace elements, characterized in that: Escherichia coli is used as the host strain, and the culture medium containing carbon source, nitrogen source, phosphorus source and trace elements is as follows: Carbon source: Glucose accounts for 15-36g / L; Nitrogen source: Amino acids account for 6-12 g / L; Phosphorus source: Phosphate accounts for 0.1-0.5 g / L; Trace elements: Iron accounts for 0.1-1.5mg / L; zinc accounts for 0.1-1.5mg / L; magnesium accounts for 0.5-11mg / L; The balance is pure water.

2. A method for preparing L-alanine, using the L-alanine described in claim 1, characterized in that: The L-alanine preparation method is as follows: S1: pre-culturing the Escherichia coli bacterial liquid containing Escherichia coli, and then preparing the culture medium, and preparing various raw materials required for providing nutrition and fermentation for Escherichia coli, that is, mixing the carbon source, nitrogen source, phosphorus source and trace elements to prepare the culture liquid; S2: preparing a stationary fermentation tank (1), mixing the Escherichia coli liquid and the culture liquid and adding them to the fermentation tank (1) for culture and fermentation; monitoring the cell density and alanine production parameters in the fermentation tank (1); S3: a stirrer (2) is provided in the fermentation tank, and a processing component (3) is provided in the stirrer. The stirrer (2) in the fermentation tank (1) is started, and the Escherichia coli liquid and the culture liquid in the fermentation tank (1) are stirred at a uniform speed by the stirrer (2) to make them fully mixed. Then, impurities floating in the mixed solution and foam generated during the stirring process are processed by the processing component (3); S4: After the fermentation is completed, alanine is extracted from the fermentation broth by an extraction method, and centrifugation and filtration are used for preliminary extraction; S5: purifying the extracted alanine by removing impurities using chromatography technology to obtain a high-purity alanine product; S6: Conduct quality inspection on the purified alanine, including purity and content indicators, to ensure that the product meets the requirements, and then package the alanine products that meet the quality requirements.

3. An L-alanine fermentation device, using the L-alanine preparation method described in claim 2, characterized in that: The fermentation device comprises a fermentation tank (1) and an agitator (2), wherein the agitator (2) comprises a stirring motor (20), a driving gear (311), a stirring column (22) and a stirring elbow (23). The stirring motor (20) is mounted on the fermentation tank (1) via a bracket, the output end of the stirring motor (20) faces downward and is connected to a control gear (21), the stirring column (22) is penetrated into the interior of the stirring column (22) along the height direction of the fermentation tank (1), and a control gear (21) is also mounted on the top of the stirring column (22), the control gear (21) on the stirring column (22) is meshed with the control gear (21) on the stirring motor (20), and the stirring elbow (23) is arranged on the stirring column (22) at equal intervals along the length direction of the stirring column (22).

4. An L-alanine fermentation device according to claim 3, characterized in that: The processing component (3) comprises a lifting frame (30) and a processing frame (31) integrally connected to the lifting frame (30), two groups of collecting sponges (32) are slidably arranged in the processing frame (31), and each group of collecting sponges (32) is slidably arranged in the processing frame (31) along the length direction of the processing frame (31), the lifting frame (30) is arranged on the stirring column (22), and the processing frame (31) is installed on the outer side wall of the lifting frame (30); A feed trough (33) and a discharge trough (34) for collecting and removing the froth generated during the preparation of the L-alanine solution are respectively provided on both sides of the processing frame (31).

5. An L-alanine fermentation device according to claim 4, characterized in that: Two groups of symmetrically distributed opening and closing doors (310) are also hinged on the feed trough (33) and the discharge trough (34) of the processing frame (31), and the two groups of opening and closing doors (310) cover the feed trough (33) and the discharge trough (34); A driving gear (311) for driving the opening and closing of the two groups of opening and closing doors (310) is disposed on the axis thereof; a plurality of the driving gears (311) are rotatably fitted to the side wall of the processing frame (31); a driving rack (312) is meshed with the driving gear (311); the driving rack (312) is slidably distributed along the height direction of the processing frame (31); a limiting frame (313) for limiting the movement of the driving rack (312) is also disposed on the side wall of the processing frame (31) close to the driving gear (311); and a reset tension spring (314) is connected between the driving rack (312) and the limiting frame (313).

6. An L-alanine fermentation device according to claim 5, characterized in that: Sawtooth structures are alternately provided on both sides of the driving rack (312), and the two driving racks (312) are respectively meshed with a plurality of driving gears (311) on both sides of the processing frame (31).

7. An L-alanine fermentation device according to claim 6, characterized in that: Each group of collecting sponges (32) further comprises a plurality of cleaning cotton pieces (35) for cleaning floating foam, and a linkage mounting plate (36) is clamped and mounted on the side walls of the plurality of cleaning cotton pieces (35), and the linkage mounting plates (36) on the two groups of cleaning cotton pieces (35) in the processing frame (31) are staggered.

8. An L-alanine fermentation device according to claim 7, characterized in that: A linkage seesaw (37) is hingedly mounted on one side of the processing frame (31); one side of the linkage seesaw (37) is slidably engaged with a linkage mounting plate (36) on one side of the processing frame (31); and the other side of the linkage seesaw (37) is engaged with a linkage mounting plate (36) on the other side of the processing frame (31).

9. An L-alanine fermentation device according to claim 3, characterized in that: The fermentation tank (1) is also provided with a linkage component (4), the linkage component (4) comprising two pressing knobs (40), the two pressing knobs (40) being slidably arranged on the fermentation tank (1), and a No. 1 rod (41) being installed at the bottom of one side of the pressing knob (40), and a start ring (42) being provided at one end of the No. 1 rod (41) away from the pressing knob (40); A second rod (43) is installed at the bottom of the pressing knob (40) on the other side. A cleaning ring (44) is provided at one end of the second rod (43) away from the pressing knob (40). A cleaning rod (45) is abutted at the bottom of the cleaning ring (44). The cleaning rod (45) is arranged on one side of the linkage rocker (37). The starting ring (42) and the cleaning ring (44) are slidably arranged inside the fermentation tank (1) along the height direction thereof.

10. An L-alanine fermentation device according to claim 4, characterized in that: Flushing pipes (46) are provided on both sides of the processing frame (31) in the width direction. The flushing pipe (46) is inserted into the stirring column (22) on the side away from the processing frame (31) and extends outward along the stirring column (22) to the outside of the fermentation tank (1). The flushing pipe (46) is connected to an external water source on the side away from the processing frame (31).

Images