L-alanine high-yield strain, extraction method and extraction equipment

Abstract

The invention relates to an L-alanine high-yield strain, an extraction method and extraction equipment, and relates to the technical field of L-alanine high-yield strains, the L-alanine high-yield strain comprises a standing extraction platform, an extraction support, an extraction guide rail and a strain extractor, the strain extractor comprises a plurality of adjacently distributed extraction filter plates, and a working inner cavity is formed in each extraction filter plate; a microwave generator is arranged in the working inner cavity of the extraction filter plate, and working outer cavities are formed in the two sides of the extraction filter plate and used for crystallizing the mixed initial strains; according to the method, the mixed solution is crystallized, so that the solution is changed from a liquid state to a solid state, at the moment, the solution can be screened, impurities and metabolites in a crystalline product are removed, and at the moment, the L-alanine high-producing strain is extracted, so that the influence of the impurities on the extraction of the L-alanine high-producing strain can be greatly avoided; the extraction efficiency of the L-alanine high-yield strain is ensured.

Description

Technical Field

[0001] The invention relates to the technical field of L-alanine high-yield strains, and in particular to an L-alanine high-yield strain, an extraction method and extraction equipment. Background Art

[0002] L-alanine is an important amino acid with wide application value in food, medicine, chemical industry and other fields. In order to improve the production efficiency of L-alanine, researchers have cultivated high-yield strains through genetic engineering technology.

[0003] High-yield strains refer to microbial strains that can produce more L-alanine during the fermentation process. By optimizing the genome of the strains, regulating related metabolic pathways and fermentation conditions, researchers have successfully constructed a series of high-yield L-alanine microbial strains.

[0004] For example, a Chinese patent with publication number CN103602609A discloses a high-yield strain for fermenting and producing L-alanine. The strain is based on "Lactobacillus delbrueckii subspecies bulgaricus" as the starting strain, and the cells are mutagenized by low-energy ion implantation, and the mutant strain is screened out by reversible inhibition assay. The growth temperature range of the strain is 35-40°C, and the optimal growth temperature is 37°C; the optimal pH value is 7.0; the slant culture medium used is MRS culture medium, and the strain uses glucose and other sugar substances as substrates to ferment and produce L-alanine under anaerobic conditions. Ultrafiltration membrane filtration and nanofiltration membrane filtration are used to remove impurities during the purification process, thereby ensuring product quality and being suitable for large-scale industrial production.

[0005] However, the above L-alanine high-yield strain extraction method has some shortcomings in actual use: 1. First, during the extraction process of the existing L-alanine high-yielding strains, some impurities and some metabolic waste gas products generated during the stirring process will be mixed in the mixed solution, resulting in poor efficiency in extracting the L-alanine high-yielding strains from the mixed solution. In addition, when extracting the L-alanine high-yielding strains from the mixed solution, the mixed solution is prone to flow, thereby increasing the difficulty of extracting the L-alanine high-yielding strains.

[0006] 2. Secondly, the existing extraction method of high-yield L-alanine strains is relatively simple and cannot guarantee the yield of high-yield L-alanine strains, resulting in the inability to mass-produce high-yield L-alanine strains, thus affecting their ultimate production capacity.

[0007] Therefore, based on the above-stated viewpoints, there is still room for improvement in the existing L-alanine high-yield strain extraction method. Summary of the invention

[0008] In order to solve the above problems, the present invention provides a high-yield L-alanine strain and an extraction method thereof, which adopts the following technical scheme: In a first aspect, the present application relates to an L-alanine high-yield strain, which is obtained by mutating Escherichia coli using a culture medium containing a mutagen, and then culturing the mutant to obtain a high-yield strain containing an L-alanine gene.

[0009] The composition of the culture medium containing the mutagen is as follows: Peptone accounts for 8-15g / L; Yeast extract accounts for 4-10g / L; Salt accounts for 2.5-10.7g / L; Glucose accounts for 1-10g / L; Amino acids account for 0.1-1.0g / L; Ethyl methanesulfonate accounts for 1-10g / L.

[0010] In a second aspect, the present application also provides a method for extracting a high-yield strain of L-alanine, comprising preparing a stationary extraction platform, on which extraction brackets are arranged at equal intervals, and extraction guide rails and a strain extractor are also arranged; three groups of the extraction guide rails are arranged on the extraction bracket, and the three groups of the extraction guide rails are distributed in a triangular shape, and the strain extractor is slidably arranged on the three groups of extraction guide rails by an electric drive; The method for extracting L-alanine high-yield strain comprises the following steps: (1): Screening out strains with high production potential from the strain library, that is, using Escherichia coli as the starting strain for preliminary culture and identification to restore the vitality of the strain; (2): Expose E. coli to a culture medium containing a mutagen for 10-180 minutes at a temperature of 30-37 degrees Celsius, and irradiate with ultraviolet light for 5 minutes to cause mutations; (3): After the E. coli is mutated, it is diluted and the diluted and mutated E. coli is inoculated into a medium rich in L-alanine for cultivation, so that the starting strain is rich in L-alanine; (4): After the initial strain is cultured, the mixed solution is crystallized, and the impurities and metabolites in the mixed solution are removed by a strain extractor; (5): Then, the mixed solution after crystallization is placed back on the observation instrument to screen out the mutant starting strain that produces high-yield L-alanine gene.

[0011] The third aspect: also relates to an L-alanine high-yield strain extraction device, the extraction device comprising an extraction platform, an extraction support, an extraction guide rail and an extractor; The strain extractor comprises a plurality of adjacently distributed extraction filter plates, hooks are arranged on both sides of the extraction filter plates for driving the extraction filter plates to be hung on three groups of extraction guide rails, and circular conveying holes are opened in the middle of the plurality of extraction filter plates for conveying the starting strains that have completed cultivation and mixed with the culture medium, a working inner cavity is opened inside the extraction filter plates, and a microwave generator is arranged in the working inner cavity of the extraction filter plates.

[0012] The two sides of the extraction filter plate are provided with working outer cavities for the mixed starting strains to be crystallized.

[0013] The working inner cavity of the extraction filter plate is also provided with ventilation grooves at equal intervals for ventilation into the working inner cavity and in linkage with the working inner cavity.

[0014] Preferably, a sealing strip for sealing is provided on one side of the extraction filter plate in the width direction, and a sealing groove for abutment of the sealing strip provided on the adjacent extraction filter plate is provided on the other side of the extraction filter plate in the width direction.

[0015] Preferably, the strain extractor also includes two symmetrically distributed guide plates, each of which is provided with a guide groove, and a guide rod is slidably installed in the guide grooves of the two guide plates. A ventilation interface and an infusion interface are provided at the bottom of the guide rod in sequence, and the tail of the ventilation interface is connected to an air pump device through a gas pipeline, and the tail of the infusion interface is connected to a storage device for storing a mixed solution through a liquid pipeline.

[0016] Preferably, the extraction platform is also provided with a pressing component for squeezing a plurality of extraction filter plates, and the pressing component includes a pressing motor, a bidirectional pressing screw, a pressing shift block, a lifting pressing block, a pressing column and two pressing plates.

[0017] The clamping motor is installed on the extraction platform through a motor seat, the bidirectional clamping screw is rotatably installed on the extraction platform through a bearing and is parallel to its length direction, the output end of the clamping motor is connected to the bidirectional clamping screw, the two clamping shift blocks are symmetrically installed on the bidirectional clamping screw by screw connection, the lifting and lowering clamping block is arranged at its upper end along the height direction of the clamping shift block, and the two sides of the lifting and lowering clamping block are connected to the clamping column, the clamping column is slidably passed through the two clamping plates, the clamping plates are provided with a clamping guide groove for the sliding of the clamping column, and the clamping plate is arranged on the extraction platform.

[0018] Preferably, a linkage component is also arranged between the clamping component on the extraction platform and the guide rod on the strain extractor, and the linkage component includes a linkage belt, a linkage threaded rod, a linkage block and a linkage vertical plate. The linkage threaded rod is rotatably installed on the extraction platform through a bearing, and the linkage threaded rod is distributed in parallel with the bidirectional clamping screw, and a linkage belt is also sleeved therebetween. The linkage block is screwed on the linkage threaded rod, and the linkage block is integrally connected to the linkage vertical plate. A bar groove is opened on the linkage vertical plate, and the guide rods slidably arranged on the two guide plates are also slidably arranged in the bar groove of the linkage vertical plate.

[0019] Preferably, the working outer cavities opened on both sides of the extraction filter plate are also attached with heat spreaders.

[0020] Preferably, a return spring is provided between the hooks of several adjacent extraction filter plates.

[0021] Preferably, the ventilation grooves in the thickness direction of the extraction filter plate are staggered, with the ventilation grooves on one side being located at the bottom of the extraction filter plate and the ventilation grooves on the other side being located at the top of the extraction filter plate.

[0022] In summary, the present application includes at least one of the following beneficial technical effects: 1. The present application crystallizes the mixed solution so that the solution changes from liquid to solid. At this time, it can be screened to remove impurities and metabolites in the crystalline product. At this time, the extraction of the L-alanine high-yielding strain can greatly avoid the influence of impurities on the extraction of the L-alanine high-yielding strain, thereby ensuring the efficiency of the extraction of the L-alanine high-yielding strain.

[0023] 2. The L-alanine high-yield strain in this application not only improves production efficiency and output, but also reduces production costs through metabolic engineering transformation and the application of fermentation technology, and has significant environmental friendliness and broad application prospects.

[0024] 3. In order to improve the efficiency of crystallization of the mixed solution and ensure that the entire equipment can automatically process the mixed solution in batches, the present application also proposes a linkage component to ensure the automatic crystallization operation of the mixed solution, while also reducing the number of external power sources and reducing the cost of the equipment. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0026] Figure 1 It is a schematic diagram of the main structure of the present invention.

[0027] Figure 2 It is a structural schematic diagram of the strain extractor of the present invention.

[0028] Figure 3It is a schematic diagram of the first-view structure between several extraction filter plates of the present invention.

[0029] Figure 4 It is a second perspective structural schematic diagram of a plurality of extraction filter plates of the present invention.

[0030] Figure 5 It is a schematic diagram of the structure between the extraction filter plate and the strain extractor of the present invention.

[0031] Figure 6 It is a first-view structural diagram between the linkage component and the strain extractor of the present invention.

[0032] Figure 7 It is a second perspective structural schematic diagram between the linkage component and the strain extractor of the present invention.

[0033] Figure 8 It is a schematic diagram of the structure between the linkage component and the clamping component of the present invention from a first perspective.

[0034] Fig. 9 It is a second perspective structural schematic diagram between the linkage component and the clamping component of the present invention.

[0035] Fig.10 It is a first-view structural schematic diagram of the blanking component of the present invention.

[0036] Fig.11 It is a second perspective structural schematic diagram of the blanking component of the present invention.

[0037] Fig.12 The present invention is a flow chart of the method for extracting L-alanine high-yielding strain.

[0038] Description of reference numerals: 1. extraction platform; 2. extraction support; 3. extraction guide rail; 4. strain extractor; 40. extraction filter plate; 41. hook; 42. delivery circular hole; 43. working inner cavity; 44. microwave generator; 45. working outer cavity; 46. ventilation slot; 400. sealing strip; 401. sealing groove; 47. guide plate; 48. guide slide; 49. guide rod; 50. ventilation interface; 51. infusion interface; 52. gas pipeline; 53. liquid pipeline; 6 , clamping parts; 60, clamping motor; 61, bidirectional clamping screw; 62, clamping shift block; 63, lifting clamping block; 64, clamping column; 65, clamping plate; 66, clamping guide groove; 7, linkage parts; 70, linkage belt; 71, linkage threaded rod; 72, linkage block; 73, linkage vertical plate; 74, strip slot; 402, heat spreader; 403, reset compression spring; 8, unloading parts; 80, unloading rod; 81, unloading scraper; 82, unloading spring; 83, unloading guide rail. DETAILED DESCRIPTION

[0039] The following is combined with Figure 1-Figure 12 This application is described in further detail.

[0040] The embodiments of the present application disclose an L-alanine high-yielding strain, an extraction method and an extraction device; it is explained that it first relates to an L-alanine high-yielding strain, and the L-alanine high-yielding strain in the present application can significantly increase the yield of L-alanine under the same unit volume.

[0041] Secondly, the present application relates to a method for extracting a high-yielding strain of L-alanine. During the extraction process of the existing high-yielding strain of L-alanine, some impurities and some metabolic waste gas products generated during the stirring process are doped in the mixed solution, resulting in poor efficiency in extracting the high-yielding strain of L-alanine from the mixed solution. In addition, when extracting the high-yielding strain of L-alanine from the mixed solution, the mixed solution is prone to flow, thereby increasing the difficulty of extracting the high-yielding strain of L-alanine. The method for extracting the high-yielding strain of L-alanine in the present application can well solve the above problems.

[0042] The invention discloses an L-alanine high-yield strain. The high-yield strain containing the L-alanine gene is obtained by mutating Escherichia coli in a culture medium containing a mutagen and then culturing the mutant.

[0043] A high-yield strain containing the L-alanine gene is prepared by a mutagenesis method to cause gene mutation, thereby obtaining a strain with specific genetic characteristics; first, Escherichia coli is selected as a starting strain, then the strain is subjected to gene mutation by a mutagen, and then a high-yield strain containing the L-alanine gene is obtained by culturing.

[0044] The components of the culture medium containing mutagen are as follows: peptone accounts for 8-15g / L; yeast extract accounts for 4-10g / L; salt accounts for 2.5-10.7g / L; glucose accounts for 1-10g / L, amino acid accounts for 0.1-1.0g / L, and ethyl methanesulfonate accounts for 1-10g / L.

[0045] Escherichia coli is one of the main microorganisms currently used for L-alanine fermentation production. Its clear genetic background, mature genetic modification technology, fast reproduction speed and short fermentation cycle make it an ideal production strain. As a high-yield strain of L-alanine, Escherichia coli has achieved commercial production through metabolic engineering and optimization of fermentation conditions, and has significant advantages in industrial applications.

[0046] In the mutagenesis method of Escherichia coli, mutagens are particularly important. Different concentrations and types of mutagens will affect the controllability and efficiency of gene mutation in Escherichia coli.

[0047] The present invention uses ethyl ethyl methanesulfonate as an important component of the mutagen. Ethyl ethyl methanesulfonate is an important alkylating agent mutagen and carcinogen, and its mutagenicity in Escherichia coli has been widely studied. Studies have shown that ethyl ethyl methanesulfonate mainly causes mutations by inducing the conversion of G:C to A:T, and this mutation accounts for 97% of all mutations induced by ethyl ethyl methanesulfonate.

[0048] Embodiment 1: The composition of the culture medium containing the mutagen is as follows: Peptone accounts for 8.9 g / L; Yeast extract accounts for 5.5 g / L; Salt accounts for 5g / L; Glucose accounts for 6g / L; Amino acids account for 0.5 g / L; Ethyl methanesulfonate accounts for 5.3 g / L.

[0049] Operation steps: First, prepare the culture medium required for E. coli mutation in advance. The pH in the culture medium is maintained at 7.2, and the culture medium is placed in a constant temperature environment of 31 degrees Celsius. Then, expose the E. coli to the culture medium and induce mutagenesis at 31 degrees Celsius for 60 minutes. After the 60-minute chemical mutagenesis, irradiate with ultraviolet light for 5 minutes. After the desired strain is obtained, the strain is fermented in a 1L fermentation tank at a constant temperature of 21 degrees Celsius for 24 hours to produce a strain containing L-alanine. The strain is then screened, and the final mutation rate of the L-alanine strain is 1x10^-6.

[0050] Embodiment 2: The composition of the culture medium containing the mutagen is as follows: Peptone accounts for 11 g / L; Yeast extract accounts for 7 g / L; Salt accounts for 9.5g / L; Glucose accounts for 9g / L; Amino acids account for 0.88 g / L; Ethyl methanesulfonate accounts for 8.5 g / L.

[0051] Operation steps: First, prepare the culture medium required for E. coli mutation in advance. The pH in the culture medium is maintained at 7.2, and the culture medium is placed in a constant temperature environment of 31 degrees Celsius. Then, expose the E. coli to the culture medium and induce mutagenesis at 31 degrees Celsius for 60 minutes. After the 60-minute chemical mutagenesis, irradiate with ultraviolet light for 5 minutes. After the desired strain is obtained, the strain is fermented in a 1L fermentation tank at a constant temperature of 21 degrees Celsius for 24 hours to produce a strain containing L-alanine. The strain is then screened, and the final mutation rate of the L-alanine strain is 1.324x10^-6.

[0052] Embodiment three: The composition of the culture medium containing the mutagen is as follows: Peptone accounts for 15g / L; Yeast extract accounts for 10 g / L; Salt accounts for 10.7 g / L; Glucose accounts for 10g / L; Amino acids account for 1.0 g / L; Ethyl methanesulfonate accounts for 10g / L.

[0053] Operation steps: First, prepare the culture medium required for E. coli mutation in advance. The pH in the culture medium is maintained at 7.2, and the culture medium is in a constant temperature environment of 31 degrees Celsius. Then, expose the E. coli to the culture medium and induce mutagenesis at 31 degrees Celsius for 60 minutes. After the 60-minute chemical mutagenesis, it is supplemented with ultraviolet irradiation for 5 minutes. After the desired strain is obtained, the strain is fermented in a 1L fermentation tank at a constant temperature of 21 degrees Celsius for 24 hours to produce a strain containing L-alanine. The strain is then screened, and the final L-alanine strain mutation rate is 1.78x10^-6.

[0054] As the content of ethyl ethyl methanesulfonate in the culture medium containing the mutagen increases within a reasonable range and nutrients are simultaneously supplied, the mutation rate of the strain containing high-yield L-alanine can be effectively increased, thereby achieving the effect of increasing the yield of the strain containing high-yield L-alanine.

[0055] Embodiment 4: Based on the above-mentioned L-alanine high-yielding strain, the present application also proposes an extraction method for the above-mentioned L-alanine high-yielding strain, which is specifically as follows: See also Fig.12 As shown, the method for extracting L-alanine from a high-yielding strain comprises the following steps: (1): First, select Escherichia coli as the starting strain, then prepare a culture medium containing a mutagen and mix ethyl methanesulfonate into the culture medium.

[0056] (2): Expose the starting strain to a culture medium containing a mutagen for 120 minutes at a temperature of 31 degrees Celsius to allow it to undergo genetic mutations.

[0057] (3): After the starting strain has completed mutation, it is diluted, and then the diluted and mutated starting strain is mixed into a medium rich in L-alanine for cultivation, so that the starting strain contains L-alanine.

[0058] (4): After the initial strain is cultured, the mixed solution is crystallized, and the impurities and metabolites in the mixed solution are removed by the strain extractor 4 to reduce the impurities in the mixed solution, thereby improving the success rate and efficiency of extracting the L-alanine high-yielding strain.

[0059] (5): Then, the mixed solution after crystallization is placed back on the observation instrument to screen out the mutant starting strain that produces high-yield L-alanine gene.

[0060] In the existing L-alanine high-yield strain extraction method, during the extraction process, a mixed solution is mixed with a medium rich in L-alanine, metabolic waste is generated during the mixing and stirring process, and some external impurities also enter the solution during the mixing. If the L-alanine high-yield strain is directly screened, the solution is prone to flow, thereby aggravating the screening of the L-alanine high-yield strain, and a large amount of impurities and metabolites are mixed in the mixed solution, which will also seriously affect the selection of the L-alanine high-yield strain, thereby greatly affecting the extraction of the L-alanine high-yield strain.

[0061] The present application crystallizes the mixed solution so that the solution changes from liquid to solid, at which time it can be screened to remove impurities and metabolites in the crystalline product. At this time, when extracting the L-alanine high-yielding strain, the influence of impurities on the extraction of the L-alanine high-yielding strain can be greatly avoided, thereby ensuring the efficiency of the extraction of the L-alanine high-yielding strain.

[0062] Embodiment five: The method for extracting a high-yield L-alanine strain in the fourth embodiment also relates to a device for extracting a high-yield L-alanine strain.

[0063] See also Figure 1 As shown, it includes a stationary extraction platform 1, an extraction bracket 2, an extraction guide rail 3 and a strain extractor 4; the extraction bracket 2 is arranged on the extraction platform 1 at equal intervals, three groups of extraction guide rails 3 are arranged on the extraction bracket 2, and the three groups of extraction guide rails 3 are distributed in a triangular shape, and the strain extractor 4 is slidably arranged on the three groups of extraction guide rails 3 in an electric manner.

[0064] It should be noted that the extraction platform 1 is set on the ground, the extraction brackets 2 are evenly distributed on the extraction platform 1, and support the extraction guide rails 3 on the extraction platform 1 to ensure that the three groups of extraction guide rails 3 can be distributed horizontally, and a number of extraction filter plates 40 are slidably installed on the three groups of extraction guide rails 3.

[0065] After the initial strain is mutated, it is diluted, and then the diluted and mutated initial strain is mixed with a medium rich in L-alanine for cultivation, so that the initial strain contains L-alanine. Then the initial strain containing L-alanine is in a mixed solution, and the mixed solution is stored in a special storage device. Then the storage device transports the mixed liquid to the strain extractor 4 through a liquid pipeline 53 for crystallization treatment. During the crystallization process, the metabolic waste in the mixed liquid is evaporated and removed. After the crystallization is completed, it is screened and filtered to remove impurities doped in the crystals, thereby obtaining the final product, and then it is screened and extracted to obtain the L-alanine high-yield strain.

[0066] See also Figure 2 , Figure 3 , Figure 4 and Figure 5 As shown, it is a schematic diagram of the structure of crystallizing a mixed solution containing a starting strain in the present application; the strain extractor 4 includes a plurality of adjacently distributed extraction filter plates 40, both sides of which are provided with hooks 41 for driving the extraction filter plates 40 to be hung on three groups of extraction guide rails 3, and the middle parts of the plurality of extraction filter plates 40 are provided with conveying circular holes 42 for conveying the starting strain that has completed the culture and mixed with the culture medium, the interior of the extraction filter plate 40 is provided with a working inner cavity 43, and a microwave generator 44 is provided in the working inner cavity 43 of the extraction filter plate 40.

[0067] Working outer chambers 45 are provided on both sides of the extraction filter plate 40 for crystallization of the mixed starting strains.

[0068] The working inner cavity 43 of the extraction filter plate 40 is provided with ventilation grooves 46 at equal intervals for ventilation into the working inner cavity 43 and in linkage therewith.

[0069] Microwave generators 44 are devices that convert electricity into microwave energy and are widely used in industries, food processing, surface treatment and science. These devices generate microwave radiation through magnetrons or solid-state microwave generators 44, using the dielectric loss characteristics of materials for heating and drying processes.

[0070] It should be noted that a working outer cavity 45 is provided outside the extraction filter plate 40, which can be filled with a mixed solution containing the starting strain and subjected to crystallization treatment by a microwave generator 44. A working inner cavity 43 is provided inside the extraction filter plate 40 for placing the microwave generator 44 to ensure that the microwave generator 44 can work stably and can also perform heat dissipation treatment on the microwave generator 44.

[0071] During the specific implementation process: first, the infusion interface 51 and the ventilation interface 50 are connected to the extraction filter plate 40 on the extraction platform 1, and then the infusion interface 51 fills a large amount of mixed solution into a plurality of extraction filter plates 40 that are in end-to-end contact, and the mixed solution passes through the delivery circular hole 42 in the middle of the extraction filter plate 40, and the mixed solution is evenly filled into the working outer cavity 45 of the extraction filter plate 40 that is in end-to-end contact, and then the microwave generator 44 is powered on to perform microwave heating treatment on the inside, until the mixed solution in the working outer cavity 45 of the extraction filter plate 40 is crystallized, and the thickness of the mixed solution after crystallization is consistent, which is convenient for the subsequent removal of impurities.

[0072] When the mixed solution containing the starting strain is injected into the working outer cavity 45 of the extraction filter plate 40, the mixed solution may leak. Therefore, the present application proposes a clamping component 6, which can ensure the sealing between several extraction filter plates 40 during the crystallization process of the mixed solution; when the mixed solution is crystallized, it can be easily separated.

[0073] See also Figure 6 and Figure 7 As shown, it is a schematic diagram of the structure of the mixed solution storage in the present application, the strain extractor 4 includes two symmetrically distributed guide plates 47, the guide plates 47 are provided with guide grooves 48, guide rods 49 are slidably installed in the guide grooves 48 of the two guide plates 47, and a ventilation interface 50 and an infusion interface 51 are sequentially arranged at the bottom of the guide rod 49, the tail of the ventilation interface 50 is connected to an air pump device through a gas pipeline 52, and the tail of the infusion interface 51 is connected to a storage device for storing the mixed solution through a liquid pipeline 53.

[0074] It should be noted that the storage device is a known structure, which can store the mixed liquid containing the starting strain, and can transport the mixed liquid to the strain extractor 4 by pressurizing.

[0075] The air pump device is a known structure, which can transport air into the extraction filter plate 40 to dissipate heat for the components in the extraction filter plate 40 .

[0076] See also Figure 8 and Fig. 9As shown, the extraction platform 1 is also provided with a pressing component 6 for pressing a plurality of extraction filter plates 40 , and the pressing component 6 includes a pressing motor 60 , a bidirectional pressing screw 61 , a pressing shift block 62 , a lifting pressing block 63 , a pressing column 64 and two pressing plates 65 .

[0077] The clamping motor 60 is installed on the extraction platform 1 through the motor seat, and the bidirectional clamping screw 61 is rotatably installed on the extraction platform 1 through a bearing and is parallel to its length direction. The output end of the clamping motor 60 is connected to the bidirectional clamping screw 61, and the two clamping shift blocks 62 are symmetrically installed on the bidirectional clamping screw 61 by screwing. The lifting and lowering clamping block 63 is arranged at the upper end of the clamping shift block 62 along the height direction of the clamping shift block 62, and the two sides of the lifting and lowering clamping block 63 are connected to the clamping column 64, and the clamping column 64 is slidably penetrated into the two clamping plates 65. The clamping plate 65 is provided with a clamping guide groove 66 for the sliding of the clamping column 64, and the clamping plate 65 is arranged on the extraction platform 1.

[0078] A return compression spring 403 is provided between the hooks 41 of a plurality of adjacent extraction filter plates 40 .

[0079] In the initial state, the plurality of return compression springs 403 press each adjacent extraction filter plate 40 , so that the plurality of extraction filter plates 40 are in a separated state.

[0080] In the initial state, the lifting and pressing block 63 is retracted to the top of the pressing and moving block 62 , and the top height of the lifting and pressing block 63 is smaller than the bottom height of the extraction filter plate 40 .

[0081] When the mixed solution needs to be crystallized, the clamping motor 60 is started, and the clamping motor 60 controls the bidirectional clamping screw 61 to rotate. At this time, the two clamping moving blocks 62 on the bidirectional clamping screw 61 approach each other. In this process, the lifting and clamping block 63 gradually moves upward along the clamping guide groove 66 of the clamping plate 65 through the clamping column 64 until the height of the lifting and clamping block 63 is higher than the bottom height of the extraction filter plate 40, and then the lifting and clamping block 63 contacts the extraction filter plate 40 and squeezes the extraction filter plate 40.

[0082] The plurality of extraction filter plates 40 are in a separated state in an initial state. When the extraction filter plates 40 at both ends of the plurality of extraction filter plates 40 are squeezed by the lifting and clamping blocks 63, the plurality of extraction filter plates 40 approach each other until the plurality of extraction filter plates 40 are tightly fitted together from head to tail. At this time, the working outer cavities 45 of the plurality of extraction filter plates 40 are also connected to each other. At this time, the mixed solution can be passed into the working outer cavity 45 for crystallization.

[0083] After crystallization is completed, the bidirectional compression screw 61 rotates in the direction to move the lifting compression block 63 away from the extraction filter plate 40 and retract to the top of the compression shift block 62 , and the extraction filter plates 40 are separated under the action of the return compression spring 403 .

[0084] Will watch Figure 4 , Figure 5 and Figure 6 In order to further improve the sealing between the several extraction filter plates 40, sealing strips 400 are arranged between the several extraction filter plates 40. When the several extraction filter plates 40 are in contact with each other, the sealing strips 400 on the extraction filter plates 40 adjacent to each other are in corresponding contact with the sealing grooves 401 to ensure the sealing of the extraction filter plates 40 after contact. Because the extraction filter plates 40 are always in a vertical state, when the mixed solution is injected into the working outer cavity 45 of the extraction filter plates 40, the sealing strips 400 can avoid the possibility of leakage of the mixed solution. However, it should be noted that the sealing strips 400 need to be replaced regularly.

[0085] The working outer chambers 45 opened on both sides of the extraction filter plate 40 are also attached with a heat spreader 402 .

[0086] The purpose is to ensure that the microwave generator 44 heats the mixed solution evenly, avoid uneven temperature of the mixed solution, and completely solve the problem of incomplete crystallization of the mixed solution.

[0087] In addition, in order to improve the efficiency of crystallization of the mixed solution and ensure that the entire equipment can automatically process the mixed solution in batches, the present application also proposes a linkage component 7 to ensure the automatic crystallization of the mixed solution, while also reducing the number of external power sources and reducing the cost of the equipment, as shown below.

[0088] Replay Figure 7 and Figure 8 As shown, a linkage component 7 is also arranged between the clamping component 6 on the extraction platform 1 and the guide rod 49 on the strain extractor 4. The linkage component 7 includes a linkage belt 70, a linkage threaded rod 71, a linkage block 72 and a linkage vertical plate 73. The linkage threaded rod 71 is rotatably installed on the extraction platform 1 through a bearing, and the linkage threaded rod 71 is parallel to the bidirectional clamping screw 61. A linkage belt 70 is also sleeved and installed between the two. The linkage block 72 is screwed on the linkage threaded rod 71, and the linkage block 72 is integrally connected with the linkage vertical plate 73. A strip groove 74 is opened on the linkage vertical plate 73, and the guide columns slidably arranged on the two guide plates 47 are also slidably arranged in the strip groove 74 of the linkage vertical plate 73.

[0089] After the clamping motor 60 is started, the bidirectional clamping screw 61 can control the linkage threaded rod 71 to rotate synchronously in the same direction through the linkage belt 70. After the linkage threaded rod 71 rotates, it drives the linkage block 72 and the linkage vertical plate 73 to approach the direction of the extraction filter plate 40. At this time, the guide rod 49 is driven by the dual action of the strip groove 74 of the linkage vertical plate 73 and the guide groove 48 of the guide plate 47 to follow the shape of the guide groove 48 of the guide plate 47 as a motion trajectory, so that the ventilation interface 50 and the infusion interface 51 on the guide rod 49 move downward to a specified height, and then the ventilation interface 50 and the infusion interface 51 are respectively inserted into the ventilation groove 46 and the conveying circular hole 42 of the extraction filter plate 40.

[0090] The ventilation interface 50 mainly introduces gas into the interior of the extraction filter plate 40 to ensure that the working inner cavity 43 in the extraction filter plate 40 maintains an air-flowing state, because the microwave generator 44 inside it will automatically generate heat after working for a long time, causing the temperature in the working inner cavity 43 of the extraction filter plate 40 to rise, and the working outer cavity 45 of the extraction filter plate 40 is also in a high temperature state. Therefore, the temperature of the working inner cavity 43 cannot be dissipated in time. After working for a long time, the microwave generator 44 will be damaged, and its service life will also be affected.

[0091] The present application continuously introduces cooling gas into the working cavity 43 of the extraction filter plate 40 , which can not only ensure that the microwave generator 44 is in a suitable working environment so that it can emit the best performance; but also prevent a large amount of dust from being deposited in the working cavity 43 .

[0092] In order to further ensure the stability and efficiency of the crystallization operation of the mixed solution containing the starting strain, the present application also proposes a feeding component 8, which can automatically feed the crystallized mixed solution to avoid the mixed solution after crystallization from sticking to the working outer cavity 45 of the extraction filter plate 40. If the mixed solution after crystallization sticks to the working outer cavity 45 of the extraction filter plate 40, it will cause the working outer cavity 45 to be closed, thereby affecting the subsequent crystallization of the mixed solution.

[0093] See also Fig.10 and Fig.11 As shown, a material unloading component 8 is also provided on the working outer cavity 45 opened on both sides of the extraction filter plate 40. The material unloading component 8 includes a material unloading rod 80, a material unloading scraper 81, a material unloading spring 82 and a material unloading guide rail 83. The material unloading rod 80 is slidably penetrated on the extraction filter plate 40 and extends vertically outward. The material unloading scraper 81 is attached to the working outer cavity 45 on both sides of the extraction filter plate 40, and the material unloading scraper 81 is connected to the material unloading rod 80. The material unloading spring 82 is sleeved on the material unloading rod 80, one side of which is connected to the material unloading rod 80 and the other side of which is connected to the extraction filter plate 40. The material unloading guide rail 83 is set on the extraction platform 1 through a bracket, and the material unloading guide rail 83 is located directly above the three groups of extraction guide rails 3 of the extraction platform 1.

[0094] It should be noted that the unloading scraper 81 is located at the top of the working outer chamber 45 in the initial state.

[0095] During the specific implementation process, after several extraction filter plates 40 complete crystallization, the extraction filter plates 40 continue to move along the extraction guide rail 3. When the discharge rod 80 on the extraction filter plate 40 contacts the discharge guide rail 83, the extraction discharge rod 80 is subjected to downward pressure. At this time, the discharge rod 80 controls the discharge scraper 81 to move downward. The discharge scraper 81 will scrape the crystals on the working outer cavity 45 of the extraction guide rail 3 to remove the crystals. The removed crystals fall into the known collection device on the extraction platform 1 for the same collection.

[0096] When the unloading rod 80 loses the squeezing of the unloading guide rail 83 , the unloading rod 80 and the unloading scraper 81 return to the initial state under the elastic force of the unloading spring 82 .

[0097] During operation: In the first step, the required mixed solution containing the starting strain and diluted after cultivation is prepared and injected into the reservoir.

[0098] Step 2: The plurality of extraction filter plates 40 on the extraction guide rail 3 are divided into a plurality of groups and moved one by one to the working area of ​​the extraction guide rail 3 in an electrically driven manner.

[0099] Step 3: Then start the clamping motor 60 and rotate the bidirectional clamping screw 61, and the two clamping moving blocks 62 on the bidirectional clamping screw 61 approach each other. During this process, the lifting and clamping block 63 gradually moves upward along the clamping guide groove 66 of the clamping plate 65 through the clamping column 64 until the height of the lifting and clamping block 63 is higher than the bottom height of the extraction filter plate 40, and then the lifting and clamping block 63 contacts the extraction filter plate 40 and squeezes the extraction filter plate 40, so that the extraction filter plates 40 are connected end to end.

[0100] Step 4: After the compacting motor 60 is started, under the control of the linkage component 7, the ventilation interface 50 and the infusion interface 51 move downward to a specified height, and then the ventilation interface 50 and the infusion interface 51 are respectively inserted into the ventilation groove 46 and the delivery circular hole 42 of the extraction filter plate 40, and the infusion interface 51 injects the mixed solution into the squeezed extraction filter plate 40 until the mixed solution fills the working outer cavity 45, and the microwave generator 44 heats and crystallizes it.

[0101] Step 5: The ventilation interface 50 allows gas to be introduced into the interior of the extraction filter plate 40 to ensure that the working inner cavity 43 in the extraction filter plate 40 maintains a state of air flow and to dissipate heat from the microwave generator 44 therein.

[0102] Step 6: After the mixed solution completes crystallization, the extraction filter plate 40 continues to move along the extraction guide rail 3. When the discharge rod 80 on the extraction filter plate 40 contacts the discharge guide rail 83, the extraction discharge rod 80 is subjected to downward pressure. At this time, the discharge rod 80 controls the discharge scraper 81 to move downward. The discharge scraper 81 will scrape the crystals on the working outer cavity 45 of the extraction guide rail 3 to remove the crystals. The removed crystals fall into the known collection device on the extraction platform 1 for the same collection.

[0103] 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. An L-alanine high-yield strain, wherein Escherichia coli is mutated by using a culture medium containing a mutagen, and then cultured to obtain a high-yield strain containing an L-alanine gene, characterized in that: The composition of the culture medium containing the mutagen is as follows: Peptone accounts for 8-15g / L; Yeast extract accounts for 4-10g / L; Salt accounts for 2.5-10.7g / L; Glucose accounts for 1-10g / L; Amino acids account for 0.1-1.0g / L; Ethyl methanesulfonate accounts for 1-10g / L.

2. A method for extracting L-alanine from a high-yielding strain, using the L-alanine high-yielding strain described in claim 1, characterized in that: A stationary extraction platform (1) is prepared, on which extraction brackets (2) are arranged at equal intervals, and extraction guide rails (3) and a strain extractor (4) are also arranged; three groups of the extraction guide rails (3) are arranged on the extraction bracket (2), and the three groups of the extraction guide rails (3) are distributed in a triangular shape, and the strain extractor (4) is slidably arranged on the three groups of extraction guide rails (3) by means of an electric drive; The method for extracting L-alanine high-yield strain comprises the following steps: (1): Screening out strains with high production potential from the strain library, that is, using Escherichia coli as the starting strain for preliminary culture and identification to restore the vitality of the strain; (2): Expose E. coli to a culture medium containing a mutagen for 10-180 minutes at a temperature of 30-37 degrees Celsius, and irradiate with ultraviolet light for 5 minutes to cause mutations; (3): After the E. coli is mutated, it is diluted and the diluted and mutated E. coli is inoculated into a medium rich in L-alanine for cultivation, so that the starting strain is rich in L-alanine; (4): After the initial strain is cultured, the mixed solution is crystallized, and the impurities and metabolites in the mixed solution are removed through the strain extractor (4); (5): Then, the mixed solution after crystallization is placed back on the observation instrument to screen out the mutant starting strain that produces high-yield L-alanine gene.

3. An L-alanine high-yield strain extraction device, using the L-alanine high-yield strain extraction method described in claim 2, characterized in that: The extraction device comprises an extraction platform (1), an extraction support (2), an extraction guide rail (3) and an extractor (4); The strain extractor (4) comprises a plurality of adjacently distributed extraction filter plates (40), hooks (41) for driving the extraction filter plates (40) to be hung on three sets of extraction guide rails (3) are provided on both sides of the extraction filter plates (40), and a plurality of the extraction filter plates (40) are provided with conveying circular holes (42) in the middle for conveying starting strains that have been cultured and mixed with a culture medium, a working inner cavity (43) is provided inside the extraction filter plates (40), and a microwave generator (44) is provided in the working inner cavity (43) of the extraction filter plates (40); The extraction filter plate (40) has two sides provided with working outer cavities (45) for crystallization of the mixed starting strains; The working inner cavity (43) of the extraction filter plate (40) is also provided with ventilation grooves (46) at equal intervals for ventilation into the working inner cavity (43) and for linkage therewith.

4. The L-alanine high-yielding strain extraction equipment according to claim 3, characterized in that: A sealing strip (400) for sealing is provided on one side of the extraction filter plate (40) in the width direction, and a sealing groove (401) for abutting against the sealing strip (400) provided on the adjacent extraction filter plate (40) is provided on the other side of the extraction filter plate (40) in the width direction.

5. The L-alanine high-yielding strain extraction equipment according to claim 3, characterized in that: The strain extractor (4) further comprises two symmetrically distributed guide plates (47), wherein the guide plates (47) are provided with guide grooves (48), wherein guide rods (49) are slidably mounted in the guide grooves (48) of the two guide plates (47), wherein the bottoms of the guide rods (49) are provided with ventilation interfaces (50) and infusion interfaces (51) in sequence, wherein the tail of the ventilation interface (50) is connected to an air pump device via a gas pipeline (52), and the tail of the infusion interface (51) is connected to a storage device for storing a mixed solution via a liquid pipeline (53).

6. The L-alanine high-yielding strain extraction equipment according to claim 3, characterized in that: The extraction platform (1) is also provided with a pressing component (6) for pressing a plurality of extraction filter plates (40), the pressing component (6) comprising a pressing motor (60), a bidirectional pressing screw (61), a pressing shift block (62), a lifting pressing block (63), a pressing column (64) and two pressing plates (65); The clamping motor (60) is mounted on the extraction platform (1) via a motor seat, the bidirectional clamping screw (61) is rotatably mounted on the extraction platform (1) via a bearing and is parallel to the length direction thereof, the output end of the clamping motor (60) is connected to the bidirectional clamping screw (61), the two clamping shift blocks (62) are symmetrically mounted on the bidirectional clamping screw (61) by screw connection, the lifting and lowering clamping block (63) is arranged at the upper end of the clamping shift block (62) along the height direction thereof, and the two sides of the lifting and lowering clamping block (63) are connected to the clamping column (64), the clamping column (64) is slidably penetrated into two clamping plates (65), the clamping plates (65) are provided with clamping guide grooves (66) for the clamping column (64) to slide, and the clamping plates (65) are arranged on the extraction platform (1).

7. The L-alanine high-yielding strain extraction equipment according to claim 6, characterized in that: A linkage component (7) is further provided between the pressing component (6) on the extraction platform (1) and the guide rod (49) on the strain extractor (4). The linkage component (7) comprises a linkage belt (70), a linkage threaded rod (71), a linkage block (72) and a linkage vertical plate (73). The linkage threaded rod (71) is rotatably mounted on the extraction platform (1) via a bearing, and the linkage threaded rod (71) is parallel to the bidirectional pressing screw rod (61). A linkage belt (70) is sleeved between the two. The linkage block (72) is screwed onto the linkage threaded rod (71), and the linkage block (72) is integrally connected to the linkage vertical plate (73). A strip-shaped slot (74) is provided on the linkage vertical plate (73), and the guide rods (49) slidably provided on the two guide plates (47) are also slidably provided in the strip-shaped slot (74) of the linkage vertical plate (73).

8. The L-alanine high-yielding strain extraction equipment according to claim 3, characterized in that: The working outer cavities (45) opened on both sides of the extraction filter plate (40) are also attached with a heat spreader (402).

9. The L-alanine high-yielding strain extraction equipment according to claim 3, characterized in that: A return compression spring (403) is provided between the hooks (41) of a plurality of adjacent extraction filter plates (40).

10. The L-alanine high-yielding strain extraction equipment according to claim 3, characterized in that: The ventilation grooves (46) in the thickness direction of the extraction filter plate (40) are staggered, with the ventilation grooves (46) on one side being located at the bottom of the extraction filter plate (40) and the ventilation grooves (46) on the other side being located at the top of the extraction filter plate (40).

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