CN112920953A - System and method for producing gamma-aminobutyric acid through large-scale microbial whole-cell transformation - Google Patents

Abstract

The invention discloses a system and a method for producing gamma-aminobutyric acid by large-scale microbial whole-cell transformation, wherein the system comprises: an aseptic workshop; a plurality of seed liquid culture shake flasks are used for receiving the seed liquid subjected to slant culture to carry out shake flask seed liquid culture; the linked discharging device is used for receiving the shake flask seed liquid for storage and distribution; a plurality of first-stage seed culture cylinders for performing first-stage seed liquid culture on the seed liquid in a shaking flask; and a plurality of fermentation units for receiving the primary seed culture solution for inoculation and fermentation. The system method adopts a biotransformation method, and the gamma-aminobutyric acid is prepared by fermenting and expressing lactic acid bacteria, so that the system method has the advantages of simple and convenient operation, mild conditions, high raw material utilization rate, high conversion rate, low separation and purification cost and the like; and the multi-layer structure of the sterile workshop is fully utilized, a continuous integrated GABA comprehensive culture fermentation system is formed, the method is safe and reliable, the operation is simple and convenient, the feeding efficiency and the production efficiency are improved, and the production period and the production cost are reduced.

Description

System and method for producing gamma-aminobutyric acid through large-scale microbial whole-cell transformation

Technical Field

The invention relates to the technical field of biological culture, in particular to a system and a method for producing gamma-aminobutyric acid through whole-cell transformation of large-scale microorganisms.

Background

Gamma-aminobutyric acid (GABA) is a natural amino acid, widely exists in animal and plant bodies, is an important inhibitory neurotransmitter in the central nervous system of mammals, is the most important neurotransmitter in brain tissues, and has various biological functions of promoting mental tranquility, increasing growth hormone secretion, resisting stress, strengthening the liver and the kidney, promoting appetite and the like. The GABA content of the plants is low, and the plants are not suitable for large-scale production. The purity of GABA products produced by the existing fermentation method is more than 20-90%, the production period is long, the production yield is low, the separation and purification of products with high purity are complex, the cost is high, and the industrial application of the products is limited, while the cost of the GABA products with high purity produced by the fermentation method in the market is high, so that the GABA products cannot meet the market demand in the future, and the industrial production cannot be realized.

Along with the fact that GABA is gradually recognized by domestic consumers, the natural raw material GABA is produced by applying the safety bacteria, is green and safe, and meets the requirements of the consumers. However, the problem of low conversion rate of products in the production of GABA by safe bacteria is generally solved because GAD (glutamate decarboxylase) gene is introduced into plasmid of lactic acid bacteria to express in the existing used genetic engineering bacteria, but the GAD secretion is insufficient due to the loss of the plasmid gene along with the increase of the passage number of the bacteria, so that the amount of glutamic acid converted into gamma-aminobutyric acid is insufficient, namely the conversion rate is low, and the accumulation of the products is influenced. In addition, the GABA is obtained by adopting a batch fermentation method in the conventional production mode, the method does not produce any action on useful substances for human beings by removing the fermentation process, repeatedly discharging, washing a tank, adding materials, sterilizing and the like, and has the defects of long production time, complicated operation, large consumption of manpower, material resources and power and the like.

Therefore, aiming at the defects that the high-purity GABA product produced by the fermentation method in the prior art has higher cost and cannot meet the market demand in the future, a comprehensive integrated device which can realize large-scale and high efficiency is urgently needed to carry out industrialized mass production of GABA, so that the large-scale finished output and market demand of GABA are met.

Disclosure of Invention

The invention aims to solve the problems in the prior art and provides a system and a method for producing gamma-aminobutyric acid through whole-cell transformation of large-scale microorganisms in industrial batch production.

In order to achieve the purpose, the invention adopts the following technical scheme:

the first aspect of the invention provides a system for producing gamma-aminobutyric acid by whole-cell transformation of scale microorganisms, which comprises:

the sterile workshop comprises a first floor, a third floor, a fourth floor, a fifth floor and a ground reservoir area arranged below the first floor; a first culture medium proportioning trough, a second culture medium proportioning trough and a third culture medium proportioning trough which are embedded in the ground reservoir area are sequentially arranged at one end of the first floor;

a plurality of seed liquid culture shake flasks, which adopt large triangular flasks, are arranged in the corresponding shaking table on the fifth floor and are used for receiving the seed liquid after slant culture to carry out shake flask seed liquid culture, the bottoms of the shake flask seed liquid culture shake flasks are connected with the hose on the gang discharging device through pipelines, and openings at the tops of the shake flask seed liquid culture shake flasks are respectively connected with the first culture medium proportioning tank through conveying hoses through a first negative pressure suction pipe;

the parallel-row type feeder is embedded on a floor slab on the fourth floor and used for receiving shake flask seed liquid cultured by the seed liquid for storage and distribution, and comprises a feeding groove and a base plate which is laterally erected on one side of the feeding groove through an extension plate, wherein a longitudinal moving mechanism used for moving the discharging valve pipe to a position corresponding to the feeding groove and a telescopic cylinder used for controlling the valve inserting plate to be opened and closed are arranged on the base plate in a sliding manner;

the plurality of primary seed culture cylinders are embedded on the floor slab of the third floor at intervals and used for receiving shake flask seed liquid distributed by the row-type feeder to perform primary seed liquid culture, the tops of the primary seed culture cylinders are respectively and correspondingly connected with the discharge valve pipes through pipelines, the tops of the primary seed culture cylinders are connected with the second culture medium distribution groove through a second negative pressure suction pipe through a conveying hose, the lower ends of the primary seed culture cylinders are positioned on the first floor, and the bottoms of the primary seed culture cylinders are respectively provided with a discharge valve; and

the fermentation units are arranged on the first floor and used for receiving the primary seed culture solution cultured by the primary seed culture cylinder for inoculation and fermentation; each group of fermentation units comprises two tank bodies; every the jar body all with its top the one-level seed culture section of thick bamboo is the correspondence and arranges, and inhales material union coupling through the third negative pressure through delivery hose respectively third culture medium batching groove and the lactic acid bacteria liquid storage tank of arranging through delivery hose connection floor four.

Furthermore, the interior of the blanking groove is divided into independent blanking bins along the length direction of the blanking groove through vertical plates, a discharging valve pipe which is vertically arranged and a valve inserting groove which is horizontally arranged are arranged at the bottom of each blanking bin, and a valve inserting plate is movably arranged in each valve inserting groove; a plate hole for communicating the discharge valve pipe with the lower storage bin is formed in the end face of one end of the valve inserting plate; and the other end of the valve inserting plate is connected with a compression spring, and the compression spring is fixed on the lower side wall of the blanking groove through a limiting frame.

Further, the longitudinal movement mechanism comprises an upper slide rail, a toothed plate, a lower slide rail which are arranged on the base plate in parallel and a driving motor which is arranged on the upper slide rail through a slide block, wherein:

the upper sliding rail, the toothed plate and the lower sliding rail are arranged in parallel along the length direction of the base plate, and the upper sliding rail and the lower sliding rail are positioned at two sides of the toothed plate;

the rotating shaft of the driving motor is connected with a walking gear through a coupler in a transmission mode, and the walking gear is in meshed connection with the toothed plate.

Further, the longitudinal movement mechanism further comprises a supporting transverse plate, a fastener arranged at one end of the supporting transverse plate and a hose fixed on the fastener, wherein:

one end of the supporting transverse plate is connected with the sliding block on the upper sliding rail;

the lower end of the hose is positioned at the discharging bin, and the upper end of the hose is connected with the seed liquid culture shake flask through the hose.

Further, the fermentation unit includes the support frame, set up in barrel mechanism on the support frame and be used for driving barrel mechanism pivoted tilting mechanism, wherein:

the supporting frame comprises an electric rotating table, a cross beam and mounting arms respectively arranged at two ends of the cross beam, and the cross beam is fixedly arranged on a rotating table top of the electric rotating table;

the cylinder mechanism comprises two tank bodies, a left fixing shaft and a right fixing shaft which are movably arranged on two side walls of the tank bodies through bearings respectively, the tank bodies are fixedly connected with the mounting arms through the left fixing shaft and the right fixing shaft which are arranged on the two side walls of the tank bodies, and the tank bodies are turned over on the mounting arms by taking the left fixing shaft and the right fixing shaft as central shafts; and

tilting mechanism includes upset motor, drive belt and overlaps respectively and locates the epaxial belt pulley of left side fixed axle, belt pulley fixed weld in on the jar body, two pass through between the belt pulley the drive belt is connected, just the drive belt is connected the upset motor, the upset motor set up in on the crossbeam.

Preferably, the support frame further comprises a base and a support table arranged on the base, wherein the two ends of the base correspond to the lower position of the tank body and are respectively provided with a discharge through hole, and the top of the support table is fixedly provided with the electric rotating table.

Further preferably, the tank body is a biconical tank body, the top of the tank body is provided with an inoculation port, and the bottom of the tank body is provided with a discharge port; an exhaust pipeline is arranged at the top of the tank body.

Further preferably, the right fixed shaft is a hollow shaft, and an air inlet pipeline extending into the tank body is arranged in the hollow shaft.

Further, the system for producing gamma-aminobutyric acid through whole-cell transformation of scale microorganisms is characterized by further comprising:

the fermentation liquor storage tank, the plate-and-frame filter press, the spray dryer and the packaging machine are sequentially arranged on the first floor, and the fermentation liquor storage tank is respectively communicated with the fermentation units on the top of the fermentation liquor storage tank through pipelines.

The second aspect of the invention provides a method for producing gamma-aminobutyric acid by whole-cell transformation of microorganisms on a large scale based on the system, which comprises the following steps:

(1) slant culture

Preparing a slant culture medium, sterilizing and cooling, and subpackaging the culture medium in a sterile test tube to prepare a test tube slant; after the test tube slant is cultured to confirm sterility, picking seed liquid and uniformly streaking in a slant culture medium, and placing the slant in an incubator for culture after streaking is finished;

controlling the culture temperature to be 30-37 ℃ and the culture time to be 8-14 h;

(2) seed liquid culture in shake flask

Preparing a seed culture medium in a first culture medium proportioning tank, inoculating the test tube slant strain cultured in the step (1) into a seed solution for culturing and shaking a flask through a first negative pressure material suction pipe under an aseptic condition after sterilization and cooling, and starting a shaking table for culturing after inoculation;

controlling the rotation speed to be 100-250 rpm and the temperature to be 30-37 ℃, and culturing for 6-12 h;

(3) first order seed culture

Weighing materials such as peptone and yeast powder according to a culture medium formula, putting the materials into a second culture medium proportioning tank, metering the volume to a metered volume, preparing a first-stage seed culture medium, sterilizing by adopting steam, and quickly cooling to 30-37 ℃ after sterilization; conveying the primary seed culture medium into a primary seed culture cylinder through a second negative pressure material suction pipe, and then distributing the shake flask seed liquid cultured in the step (2) into the corresponding primary seed culture cylinder by adopting a row type feeder;

the temperature is controlled to be 30-37 ℃, the rotating speed is 100-280 rpm, and the ventilation volume is 50-200 m in the fermentation process³H, the tank pressure is 0.03-0.06 MPa, and the microscopic examination is free of mixed bacteria; sampling in the fermentation process, detecting physical and chemical indexes and sterility conditions, and culturing in a seed solution for 6-12 h;

(4) fermentation culture

Weighing materials such as peptone and yeast powder according to a culture medium formula, putting the materials into a second culture medium proportioning tank, fixing the volume to a measured volume, sterilizing by using steam, and quickly cooling to a culture temperature after the sterilization is finished; conveying the primary seed culture medium to a corresponding tank body through a third negative pressure material suction pipe, controlling the temperature, the rotating speed, the ventilation volume, the tank pressure and other control points of the fermentation tank to meet the process requirements, and then respectively inoculating the primary seed culture solution cultured in the step (3) and the lactic acid bacteria in the lactic acid bacteria liquid storage tank, wherein the addition amount of the lactic acid bacteria is 20-100 mg/L;

the temperature is controlled to be 25-37 ℃, the rotating speed is 50-130 rpm, and the ventilation volume is 500-2000 m in the fermentation process³H, the tank pressure is 0.03-0.06 MPa, and after fermentation is carried out for 25-40 h, glutamic acid is added for continuous fermentation for 10-24 h;

(5) can placing and filter pressing

After the fermentation in the step (4) is finished, conveying the fermentation liquor to a fermentation liquor storage tank, adding 0.5-3% of filter aid into the fermentation liquor, stirring for 15-30 min, and conveying to a plate-and-frame filter press for filter pressing treatment;

(6) spray drying

Sending the filtrate subjected to filter pressing in the step (5) into a spray dryer for spray drying, wherein the air inlet temperature is controlled to be 130-180 ℃ and the air outlet temperature is controlled to be 65-95 ℃ during spraying, and the air inlet and outlet temperatures are required to be adjusted if the flowability of sprayed powder is poor and the sprayed powder is stuck to a tower;

(6) sieving and packaging

And (5) sieving the dried powder in the step (5), and packaging and labeling by using a packaging machine according to the specification.

By adopting the technical scheme, compared with the prior art, the invention has the following technical effects:

(1) the multi-layer structure of the sterile workshop is fully utilized, the processes of culture medium preparation, slant culture, shake flask seed culture, primary seed culture, fermentation culture, filtration, drying, packaging and the like are reasonably arranged in the sterile workshop, a GABA comprehensive culture fermentation system and a production process which are continuously integrated from top to bottom are formed, the production efficiency is more than 10 times that of the existing batch fermentation production, and the GABA comprehensive culture fermentation system is safe, reliable, simple and convenient to operate, improves the feeding efficiency and the production efficiency, reduces the production period and the production cost, and realizes large-scale finished GABA output; and only a small amount of workers are needed to operate, so that the labor cost is reduced, and time and labor are saved;

(2) the GABA is produced by adopting a biotransformation method, and an intelligence protection factor lactobacillus for preventing plasmid gene loss is added in the GABA fermentation process, so that the propagation rate of genetic engineering can be reduced to a certain extent by the lactobacillus, the probability of plasmid gene loss due to more passages is reduced, and the engineering bacteria can express more target enzymes for subsequent transformation; the GABA expressed by lactobacillus fermentation is safe and harmless, the production period is short, and the conversion rate is high;

(3) the multiple discharging bins of the row type discharging device are used for performing transitional storage on the seed culture solution cultured by the seed solution in the upper layer and subjected to shake flask culture, the seed culture solution can be used for batch storage of different batches of seed culture solutions, the application range is expanded, and the use is flexible and convenient; the longitudinal moving mechanism moves transversely and synchronously drives the discharge valve pipe corresponding to the bottom of the lower storage bin to move, and the telescopic air cylinder is matched to flexibly control the opening and closing of the valve inserting plate so as to realize that the lower storage bin transfers seed culture liquid into a corresponding primary seed culture cylinder below to perform primary seed liquid culture;

(4) the first culture medium proportioning tank, the second culture medium proportioning tank, the third culture medium proportioning tank and the bacterial liquid tank are arranged in the ground reservoir area below the first floor, so that the field configuration of a seed culture medium, a first-level seed culture medium and a fermentation culture medium and the supply of protective factor lactic acid bacteria are respectively realized, the supplement of additives such as each culture medium and lactic acid bacteria is realized through a corresponding negative pressure material suction pipe, the spatial structure of the first floor and the ground reservoir area is fully utilized as a raw material for storage, each raw material is not required to be transferred to a corresponding floor, the operation is simple and convenient, and the feeding efficiency is greatly improved;

(5) the fermentation unit with a double-cylinder structure is adopted in the fermentation process, the cell fermentation liquor in the cell culture cylinder is stirred in a rolling mode, the rolling in the fermentation culture process is mild, the damage of the existing stirring type fermentation culture to cells in the fermentation liquor is reduced, no foam is generated in the turning process, the higher cell fermentation density can be obtained, and the cell activity is not influenced; after fermentation is finished, the tank bodies obliquely fixed on the left side and the right side can be controlled by the support frame to carry out centrifugal motion under the action of the electric rotating table, so that a fermentation product in the tank bodies is centrifugally separated from a culture medium, fermentation liquor is conveniently collected, and subsequent filtering and drying efficiency is improved;

(6) the adopted biotransformation method, namely the whole-cell transformation method, has the advantages of simple and convenient operation, mild conditions, high utilization rate of raw materials, high transformation rate, low separation and purification cost and the like, the gamma-aminobutyric acid is prepared by fermenting and expressing lactic acid bacteria, has no toxic or side effect, is mainly applied to the feed additive industry, can be widely applied to various stresses such as material transfer, field crossing, transportation, weather change and the like, and is more and more favored in the market.

Drawings

FIG. 1 is a schematic diagram of the overall structure of a system for producing gamma-aminobutyric acid by whole-cell transformation of microorganisms in a large scale according to the present invention;

FIG. 2 is a schematic top view of a row-type feeder in a system for large-scale production of gamma-aminobutyric acid by whole-cell transformation of microorganisms according to the present invention;

FIG. 3 is a schematic bottom view of a row-type material feeder in a system for large-scale production of gamma-aminobutyric acid by whole-cell transformation of microorganisms according to the present invention;

FIG. 4 is a schematic diagram of the rear view structure of a row-type feeder in a system for large-scale production of gamma-aminobutyric acid by whole-cell transformation of microorganisms according to the present invention;

FIG. 5 is a schematic diagram of the structure of the row-type material feeder in the system for large-scale production of gamma-aminobutyric acid by whole cell transformation of microorganisms according to the present invention;

FIG. 6 is a schematic side view of a row-type discharger in a system for large-scale production of gamma-aminobutyric acid by whole-cell transformation of microorganisms according to the present invention;

FIG. 7 is a schematic diagram of the overall structure of a row-type feeder in a system for large-scale production of gamma-aminobutyric acid by whole-cell transformation of microorganisms according to the present invention;

FIG. 8 is a schematic diagram showing the overall structure of a fermentation unit in a system for producing gamma-aminobutyric acid through whole-cell transformation of microorganisms in a large scale according to the present invention;

FIG. 9 is a schematic diagram of a specific structure of a fermentation unit in a system for producing gamma-aminobutyric acid by large-scale microbial whole-cell transformation according to the present invention;

FIG. 10 is a schematic diagram of a partial enlarged structure of a part A of a fermentation unit in a system for producing gamma-aminobutyric acid through whole-cell transformation of microorganisms in a large scale according to the present invention;

FIG. 11 is a schematic diagram of the rear view structure of a fermentation unit in a system for producing gamma-aminobutyric acid through the whole-cell transformation of microorganisms in a large scale according to the present invention;

FIG. 12 is a schematic diagram showing a partial enlarged structure of a fermentation unit B in a system for producing gamma-aminobutyric acid through whole-cell transformation of microorganisms in a large scale according to the present invention;

FIG. 13 is a schematic diagram of a fermentation unit in a reversed state in a system for producing gamma-aminobutyric acid through whole-cell transformation of microorganisms in a large scale according to the present invention;

FIG. 14 is a schematic diagram of the layout structure of a fermentation liquid storage tank, a plate-and-frame filter press, a spray dryer and a packaging machine in the system for producing gamma-aminobutyric acid through large-scale microbial whole-cell transformation according to the present invention;

FIG. 15 is a schematic process flow diagram of a large-scale microbial whole-cell transformation method for producing gamma-aminobutyric acid according to the present invention;

wherein the reference symbols are:

100-sterile workshop, 101-floor one, 102-floor one, 103-floor three, 104-floor four, 105-floor five, 106-reservoir area, 107-first culture medium proportioning tank, 108-first negative pressure suction pipe, 109-second culture medium proportioning tank, 110-second negative pressure suction pipe, 111-third culture medium proportioning tank, 112-third negative pressure suction pipe, 113-bacteria liquid tank, 114-fourth pressure suction pipe, 115-lactic acid bacteria liquid storage tank, 116-negative pressure suction pump;

300-row blanking device, 301-blanking groove, 302-blanking bin, 303-extension plate, 304-supporting transverse plate, 305-fastener, 306-hose, 307-discharging valve pipe, 308-valve inserting groove, 309-valve inserting plate and 310-plate hole; 311-a buffer block, 312-a mandril, 313-a compression spring, 314-a limit frame, 315-a vertical plate, 316-an ear plate, 317-a reset spring, 318-a base plate, 319-an upper sliding rail, 320-a toothed plate, 321-a driving motor, 322-a coupler, 323-a walking gear, 324-a lower sliding rail, 325-a fixed plate with holes, 326-a supporting seat and 327-a telescopic cylinder;

500-fermentation unit, 510-support frame, 511-base, 512-support table, 513-electric rotating table, 514-beam, 515-installation arm; 520-cylinder mechanism, 521-cylinder body, 522-inoculation port, 523-discharge port, 524-exhaust pipeline, 525-left fixed shaft, 526-right fixed shaft and 527-air inlet pipeline; 530-a turnover mechanism, 531-a turnover motor, 532-a transmission belt, 533-a belt pulley, 534-a limiting hole, 535-a limiting plate, 536-a limiting bolt and 537-a tension pulley;

200-culturing and shaking the seeds in a liquid culture medium; 400-first-level seed culture cylinder; 600-fermentation liquor storage tank, 601-hopper; 700-plate and frame filter press; 800-spray dryer; 900-packaging machine.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.

All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

In order to overcome the defects of long production period, complicated operation, large consumption of manpower, material resources and power and the like in the existing batch fermentation method for producing GABA products, a system for producing gamma-aminobutyric acid through large-scale microbial whole-cell transformation is provided, and the system is a large-scale and efficient comprehensive integrated device capable of realizing industrial mass production of GABA and can meet the large-scale finished output and market requirements of GABA.

As shown in fig. 1 and fig. 14, the system for producing gamma-aminobutyric acid by whole-cell transformation of scale microorganisms specifically comprises: the system comprises an aseptic workshop 100 with a multilayer structure, a plurality of seed liquid culture shake flasks 200, a row-type feeder 300, a plurality of first-stage seed culture cylinders 400, a plurality of fermentation units 500, a fermentation liquid storage tank 600, a plate-and-frame filter press 700, a spray dryer 800 and a packaging machine 900, wherein the seed liquid culture shake flasks 200, the row-type feeder 300, the first-stage seed culture cylinders 400 and the fermentation units 500 are reasonably arranged on corresponding floors of the aseptic workshop 100 and are used for seed culture and fermentation.

In the present embodiment, as shown in fig. 1, the sterile workshop 100 has a multi-level structure, which at least includes a first floor 101, a first floor 102, a third floor 103, a fourth floor 104, a fifth floor 105, and a warehouse area 106 disposed below the first floor 101; one end of the first floor 101 is sequentially provided with a first culture medium batching groove 107, a second culture medium batching groove 109 and a third culture medium batching groove 111 which are embedded in the ground reservoir area 106, a bacteria liquid groove 113 is embedded in the ground reservoir area 106 at the other end, the negative pressure material suction pipes are respectively communicated to corresponding floors through corresponding negative pressure material suction pipes, negative pressure material suction pumps 116 are arranged on the negative pressure material suction pipes, and the conveyance of lactic acid bacteria in the culture medium or the bacteria liquid groove 113 in the first culture medium batching groove 107, the second culture medium batching groove 109 and the third culture medium batching groove 111 is realized through the negative pressure material suction pumps 116. Through set up first culture medium batching groove 107, second culture medium batching groove 109, third culture medium batching groove 111 and fungus liquid groove 113 in floor first 101 sets up below ground storehouse district 106, realize the on-the-spot configuration of seed culture medium, the first order seed culture medium, the supply of protection factor lactic acid bacteria respectively, and inhale the material pipe through corresponding negative pressure and realize the replenishment of additives such as each culture medium and lactic acid bacteria, make full use of floor first 101 and ground storehouse district 106's spatial structure as the raw materials storage, need not to transfer each raw materials to corresponding floor, easy operation is convenient, the efficiency of the material of puting in has been improved greatly.

In addition, the first medium ingredient tank 107, the second medium ingredient tank 109, the third medium ingredient tank 111 and the bacteria liquid tank 113 can be used for conveying the cleaning liquid to the seed liquid culture shake flask 200, the tandem type feeder 300, the first-stage seed culture cylinder 400 and the fermentation unit 500 through the corresponding negative pressure suction pipes to perform corresponding cleaning operation, so as to prevent cross contamination.

In this embodiment, as shown in fig. 1, a plurality of seed liquid culture shake flasks 200 are large-sized triangular flasks, each of which is equipped with a corresponding stirring paddle, a ventilation pipe, and a heating jacket, the large-sized triangular flasks are disposed in the corresponding shaking table on floor five 105, and are used for receiving the seed liquid after slant culture to perform seed liquid culture, the bottoms of the large-sized triangular flasks are connected in series through a pipeline to a hose 306 of a feeder 300, and the top openings of the large-sized triangular flasks are connected to a first culture medium dispensing tank 107 through a first negative pressure suction pipe 108 via a delivery hose. It is worth noting that the slant culture process of the system is completed on floor five 105, which is convenient for inoculating the slant strain of the test tube to the seed solution for culturing and shaking the flask, and starting the shaking table for culturing after inoculation.

In this embodiment, as shown in fig. 1, fig. 2, fig. 3 and fig. 4, the row-by-row type feeder 300 is embedded on the floor of the fourth floor 104, the discharge valve pipe 307 at the bottom is located at the top of the third floor 103, and the row-by-row type feeder 300 is used for receiving the seed liquid of the shake flask cultured by the seed liquid culture shake flask 200 for storage and distribution. The row type blanking device 300 mainly comprises a blanking groove 301 and a base plate 318 which is laterally erected on one side of the blanking groove 301 through an extension plate 303, wherein a longitudinal moving mechanism for moving a discharging valve pipe 307 to a position corresponding to the blanking groove 301 and a telescopic cylinder 327 for controlling the opening and closing of an inserting valve plate 309 are arranged on the base plate 318 in a sliding manner; the telescopic cylinder 327 is movable left and right with the longitudinal movement mechanism to control the corresponding valve insertion plate 309.

In this embodiment, as shown in fig. 1, a plurality of first-stage seed culture cylinders 400 are embedded at intervals on the floor of the third floor 103, and are respectively configured with corresponding devices such as a stirring paddle, a ventilation pipe and a heating jacket for receiving the shake flask seed liquid distributed by the parallel feeder 300 to perform first-stage seed liquid culture, the tops of the first-stage seed culture cylinders are respectively and correspondingly connected with the discharge valve pipes 307 through pipelines, the tops of the first-stage seed culture cylinders are connected with the second culture medium distribution tank 109 through the second negative pressure suction pipe 110 through the delivery hose, the lower ends of the first-stage seed culture cylinders are located on the first floor 102, and the bottoms of the first-stage seed.

In this embodiment, as shown in fig. 1, 8 and 9, a plurality of fermentation units 500 are arranged on the floor of the first floor 102 at intervals, and each group of fermentation units 500 corresponds to one primary seed culture cylinder 400 or two primary seed culture cylinders 400 above the fermentation unit as required, and is configured to receive the primary seed culture solution cultured by the corresponding primary seed culture cylinder 400 for inoculation and fermentation; each set of fermentation unit 500 comprises two tank bodies 521, and each tank body 521 is respectively provided with a corresponding breather pipe, a heating sleeve and other devices; each tank 521 is arranged corresponding to the first-stage seed culture cylinder 400 on the top, and is connected with the third culture medium distribution groove 111 through the third negative pressure suction pipe 112 through the delivery hose and connected with the lactobacillus liquid storage tank 115 arranged on the floor four 104 through the delivery hose. The lactobacillus liquid storage tank 115 is used for adding an intelligent protection factor for preventing the loss of the plasmid gene, the intelligent protection factor adopts lactobacillus, and the principle is to reduce the propagation rate of genetic engineering to a certain extent, reduce the probability of losing the plasmid gene due to more passages and allow the engineering bacteria to express more target enzyme for subsequent transformation.

As shown in fig. 1 and 14, the system for producing gamma-aminobutyric acid by whole-cell transformation of scale microorganisms further comprises: the fermentation liquor storage tank 600, the plate-and-frame filter press 700, the spray dryer 800 and the packaging machine 900 are sequentially arranged on the first floor 101, and the fermentation liquor storage tank 600 is respectively communicated with the fermentation units 500 on the top of the fermentation liquor storage tank through pipelines. After fermentation is finished by adopting the fermentation liquor storage tank 600, the fermentation liquor is conveyed to the fermentation liquor storage tank 600, 0.5-3% of filter aid is added into the fermentation liquor storage tank 600, the mixture is stirred for 15-30 min and then conveyed to a plate-and-frame filter press 700 for filter pressing, and the filtrate is conveyed to a spray dryer 800 for spray drying.

With reference to fig. 1, the system for producing γ -aminobutyric acid by large-scale whole-cell transformation of microorganisms provided in this embodiment makes full use of the multi-layer structure of the aseptic workshop, and reasonably arranges the processes of preparation of culture medium, slant culture, seed culture by shaking, primary seed culture, fermentation culture, filtration, drying, packaging and the like in the aseptic workshop to form a top-down continuous integrated GABA integrated culture fermentation system and production process, wherein the production efficiency is 10 times or more of that of the existing batch fermentation production, and the system is safe, reliable, simple and convenient to operate, improves the feeding efficiency and production efficiency, reduces the production period and production cost, and realizes large-scale finished GABA output; and only a small amount of workers are needed to operate, so that the labor cost is reduced, and time and labor are saved.

The system for producing gamma-aminobutyric acid by large-scale microbial whole-cell transformation adopts a biotransformation method to produce GABA, and an intelligence protection factor lactobacillus for preventing plasmid genes from being lost is added in the GABA fermentation process through the lactobacillus liquid storage tank 115, so that the propagation rate of genetic engineering can be reduced to a certain extent through the lactobacillus, the probability that the plasmid genes are lost due to more passages is reduced, and the engineering bacteria can express more target enzymes to perform subsequent transformation procedures; and GABA expressed by lactobacillus fermentation is safe and harmless, the production period is short, and the conversion rate is high.

Example 2

Unlike the above embodiment 1, as shown in fig. 1, fig. 2, fig. 3 and fig. 7, this embodiment provides a row-type feeder 300 which is flexible and convenient to use and can be used for batch storage and corresponding delivery of different batches of shake flask seed liquid to corresponding seed culture pots 400.

In this embodiment, as shown in fig. 2, 5, 6 and 7, the tandem type blanking device 300 includes a blanking slot 301 and a base plate 318 standing on one side of the blanking slot 301 through an extending plate 303, the inside of the blanking slot 301 is divided into 6 independent blanking bins 302 through vertical plates 315 along the length direction thereof, and the cross section of each blanking bin 302 is in a V-shaped structure for facilitating blanking; the bottom of each lower bin 302 is provided with a vertically arranged discharge valve pipe 307 and a horizontally arranged valve insertion groove 308, and an valve insertion plate 309 is movably arranged in the valve insertion groove 308; a plate hole 310 for communicating the discharge valve pipe 307 with the blanking bin 302 is formed in the end face of one end of the valve inserting plate 309; the other end of the valve inserting plate 309 is connected with a compression spring 313, and the compression spring 313 is fixed on the lower side wall of the blanking groove 301 through a limiting frame 314.

In the practical use process, insert valve plate 309 receives inboard thrust all the time under compression spring 313's effect for insert the one end that does not have the trompil on valve plate 309 all the time and be located between discharge valve pipe 307 and feed bin 302, block discharge valve pipe 307 and feed bin 302 intercommunication, make shake flask seed liquid in feed bin 302 can't get into in the first order seed culture section of thick bamboo 400 through discharge valve pipe 307. When the valve inserting plate 309 is pushed by the telescopic cylinder 327, the compression spring 313 is compressed, the plate hole 310 at one end of the valve inserting plate 309 is pushed to the position of the discharge valve pipe 307, and the feed discharging bin 302 and the discharge valve pipe 307 are communicated, so that the finished feed in the feed discharging bin 302 enters the primary seed culture cylinder 400 through the plate hole 310 and the discharge valve pipe 307, and quantitative shake flask seed liquid is put into each corresponding primary seed culture cylinder 400.

In this embodiment, as shown in fig. 3, 4, 6 and 7, the longitudinal moving mechanism includes an upper slide rail 319, a tooth plate 320, a lower slide rail 324, which are arranged on the base plate 318 in parallel, and a driving motor 321 arranged on the upper slide rail 319 through a slider, wherein: the upper slide rail 319, the tooth plate 320 and the lower slide rail 324 are arranged in parallel along the length direction of the base plate 318, and the upper slide rail 319 and the lower slide rail 324 are located at two side positions of the tooth plate 320; a rotating shaft of the driving motor 321 is in transmission connection with a walking gear 323 through a coupler 322, and the walking gear 323 is in meshing connection with the toothed plate 320; the telescopic cylinder 327 is fixed on the perforated sliding plate 325, the perforated sliding plate 325 is slidably arranged on the lower sliding rail 324, a supporting seat 326 on the perforated sliding plate 325 is connected with one end of a traveling gear 323 through a bearing, the other end of the traveling gear 323 is connected with a rotating shaft of a driving motor 321 through a coupler 322, the driving motor 321 drives the traveling gear 323 to move left and right along the toothed plate 320 through the coupler 322, and therefore the supporting transverse plate 304 and the hose 306 arranged on the supporting transverse plate 304 move left and right to the corresponding blanking bin 302.

In the present embodiment, as shown in fig. 2, 6 and 7, the longitudinal moving mechanism further includes a supporting cross plate 304, a fastener 305 disposed at one end of the supporting cross plate 304, and a hose 306 fixed to the fastener 305, wherein: one end of the supporting transverse plate 304 is connected with the slide block on the upper slide rail 319; the lower end of the hose 306 is located at the lower bin 302, and the upper end is connected with the seed liquid culture shake flask 200 through the hose.

In this embodiment, the system for producing γ -aminobutyric acid through whole-cell transformation of microorganisms in large scale employs the coupled discharging device 300 shown in fig. 2, fig. 3, fig. 4, fig. 5, fig. 6 and fig. 7, and uses a plurality of discharging bins 302 of the coupled discharging device 300 to perform transitional storage on the seed culture solution cultured in the previous seed solution culture shake flask 200, which can be used for batch storage of different batches of seed culture solutions, thereby expanding the application range and being flexible and convenient to use; and the discharge valve pipe 307 corresponding to the bottom of the lower storage bin 302 is synchronously driven to move through the transverse movement of the longitudinal moving mechanism, and the telescopic cylinder 327 is matched to flexibly control the opening and closing of the valve inserting plate 309 so as to realize that the lower storage bin 302 transfers the seed culture solution into the corresponding primary seed culture cylinder 400 downwards to perform primary seed liquid culture of the next procedure.

Example 3

Unlike the above embodiment 1, as shown in fig. 1 and 8, the present embodiment provides a fermentation unit 500 with novel structural design, low energy consumption and high fermentation effect. The fermentation unit 500 mainly comprises three parts, including a support frame 510, a cylinder mechanism 520 arranged on the support frame 510 and a turnover mechanism 530 used for driving the cylinder mechanism 520 to rotate, wherein the cylinder mechanism 520 is combined with a turnover motor 531 and an electric rotating platform 513, so that the self-turnover stirring and centrifugal separation functions of the left tank 521 and the right tank 521 are realized, the operation is simple and convenient, the operation is reliable, large-scale fermentation culture can be stably carried out on fermentation liquor, and the fermentation unit has good and industrial application prospects.

In the present embodiment, as shown in fig. 9 and 11, the supporting frame 510 includes an electric rotating platform 513, a cross beam 514, and mounting arms 515 respectively disposed at two ends of the cross beam 514, and the cross beam 514 is fixedly disposed on a rotating table top of the electric rotating platform 513; the electric rotating platform 513 is a high-efficiency electric rotating platform for controlling the fermentation product in the tank 521 to centrifugally separate from the culture medium at a rotation speed of 800-. In addition, the supporting frame 510 further includes a base 511 and a supporting platform 512 disposed on the base 511, wherein the two ends of the base 511 are respectively provided with a discharging through hole 516 corresponding to the lower position of the tank 521, and the top of the supporting platform 512 is fixedly provided with an electric rotating platform 513.

In this embodiment, as shown in fig. 9, 10, 11 and 12, the cylinder mechanism 520 includes two tank 521, and a left fixing shaft 525 and a right fixing shaft 526 movably disposed on two side walls of the tank 521 through bearings, respectively, the tank 521 is fixedly connected to the mounting arm 515 through the left fixing shaft 525 and the right fixing shaft 526 disposed on the two side walls, and the tank 521 is turned over on the mounting arm 515 with the left fixing shaft 525 and the right fixing shaft 526 as central axes, so as to stir and mix the fermentation liquid of the tank 521, reduce the damage to the microorganism in the fermentation liquid, and the culture medium does not generate foam, and can be used for large-scale whole cell transformation fermentation culture of the microorganism.

In this embodiment, as shown in fig. 9, 10, 11 and 12, the flipping mechanism 530 includes a flipping motor 531, a transmission belt 532 and belt pulleys 533 respectively sleeved on the left fixed shaft 525, the belt pulleys 533 are fixedly welded on the tank 521, the two belt pulleys 533 are connected by the transmission belt 532, the transmission belt 532 is connected to the flipping motor 531, and the flipping motor 531 is disposed on the cross beam 514. The tank 521 is a biconical tank, the top of which is provided with an inoculation port 522 and the bottom of which is provided with a discharge port 523; an exhaust pipe 524 is arranged at the top of the tank 521.

In this embodiment, as shown in fig. 9, 10, 11 and 12, the tank 521 is a double-cone tank, the outer wall of which is provided with an electric heating jacket, the top of which is provided with an inoculation opening 202, and the bottom of which is provided with a discharge opening 203. An exhaust duct 204 is arranged at the top of the tank 521, the right fixed shaft 526 is a hollow shaft, an air inlet duct 527 extending into the tank 521 is arranged in the hollow shaft, and air is supplied into the tank 521 through the air inlet duct 527.

In the present embodiment, as shown in fig. 9, 10, 11 and 12, the end surface of the pulley 533 is provided with a plurality of circularly distributed limiting holes 534. Correspondingly, a limiting plate 535 with a perforation arranged corresponding to the belt pulley 533 is welded at one end of the mounting arm 515, and the limiting plate 535 and the belt pulley 533 are in limiting connection through a limiting bolt 536. The tank 521 is fixed to the mounting arm 515 by a limit pin 536 inserted into the through holes of the limit hole 534 and the limit plate 535.

In this embodiment, as shown in fig. 13, the turnover mechanism 530 further includes a tension wheel 537 disposed on the cross beam 514, and the tension wheel 537 is disposed in cooperation with the transmission belt 532 to regulate the transmission belt 532.

After the fermentation unit 500 is adopted to complete fermentation, the overturning motor 531 is turned off at this time, the two tank bodies 521 are inclined inwards by 30-60 degrees, the tank bodies 521 are obliquely fixed on the mounting arms 515 by the limiting bolts 536, the electric rotating table 513 is started to rotate at a high speed of 1000r/min and operate for 10min, fermentation liquor in the tank bodies 521 is subjected to centrifugal separation, the next separated sediment layer is discharged from the bottom discharge port 523 of the tank bodies 521, and then the fermentation liquor separated at the upper layer is transferred from the bottom discharge port 523 of the tank bodies 521 to the fermentation liquor storage tank 600 below through the hose for subsequent treatment. After discharging is finished, the limiting bolt 536 is removed to reset the tank 521, and after cleaning, the overturning motor 531 is continuously started to perform the next fermentation process.

The fermentation unit with the double-cylinder structure is adopted in the fermentation process of a system for producing gamma-aminobutyric acid by large-scale microbial whole-cell conversion, the cell fermentation liquid of the cell culture cylinder is stirred in a rolling mode, the rolling in the fermentation culture process is mild, the damage of the existing stirring type fermentation culture to cells in the fermentation liquid is reduced, no foam is generated in the overturning process, the higher cell fermentation density can be obtained, and the cell activity is not influenced; after fermentation is finished, the tank body which is obliquely fixed on the left side and the right side can be controlled by the support frame to carry out centrifugal motion under the action of the electric rotating table, so that a fermentation product in the tank body is centrifugally separated from a culture medium, the fermentation product is accumulated at the bottom of the tank body, fermentation liquor is conveniently collected, and subsequent filtering and drying efficiency is improved.

Example 4

Based on the system for producing gamma-aminobutyric acid through whole-cell transformation of microorganisms in scale according to the above embodiment, as shown in fig. 13, a method for producing gamma-aminobutyric acid through whole-cell transformation of microorganisms in scale based on the system is provided, which specifically includes the following steps:

(1) slant culture

Preparing a slant culture medium, sterilizing and cooling, and subpackaging the culture medium in a sterile test tube to prepare a test tube slant; after the test tube slant is cultured to confirm sterility, picking seed liquid and uniformly streaking in a slant culture medium, and placing the slant in an incubator for culture after streaking is finished;

controlling the culture temperature to be 30-37 ℃ and the culture time to be 8-14 h;

(2) seed liquid culture in shake flask

Preparing a seed culture medium in a first culture medium proportioning tank, inoculating the test tube slant strain cultured in the step (1) into a seed solution for culturing and shaking a flask through a first negative pressure material suction pipe under an aseptic condition after sterilization and cooling, and starting a shaking table for culturing after inoculation;

controlling the rotation speed to be 100-250 rpm and the temperature to be 30-37 ℃, and culturing for 6-12 h;

(3) first order seed culture

Weighing materials such as peptone and yeast powder according to a culture medium formula, putting the materials into a second culture medium proportioning tank, metering the volume to a metered volume, preparing a first-stage seed culture medium, sterilizing by adopting steam, and quickly cooling to 30-37 ℃ after sterilization; conveying the primary seed culture medium into a primary seed culture cylinder through a second negative pressure material suction pipe, and then distributing the shake flask seed liquid cultured in the step (2) into the corresponding primary seed culture cylinder by adopting a row type feeder;

the temperature is controlled to be 30-37 ℃, the rotating speed is 100-280 rpm, and the ventilation volume is 50-200 m in the fermentation process³H, the tank pressure is 0.03-0.06 MPa, and the microscopic examination is free of mixed bacteria; sampling in the fermentation process, detecting physical and chemical indexes and sterility conditions, and culturing in a seed solution for 6-12 h;

(4) fermentation culture

Weighing materials such as peptone and yeast powder according to a culture medium formula, putting the materials into a second culture medium proportioning tank, fixing the volume to a measured volume, sterilizing by using steam, and quickly cooling to a culture temperature after the sterilization is finished; conveying the primary seed culture medium to a corresponding tank body through a third negative pressure material suction pipe, controlling the temperature, the rotating speed, the ventilation volume, the tank pressure and other control points of the fermentation tank to meet the process requirements, and then respectively inoculating the primary seed culture solution cultured in the step (3) and the lactic acid bacteria in the lactic acid bacteria liquid storage tank;

the temperature is controlled to be 25-37 ℃, the rotating speed is 50-130 rpm, and the ventilation volume is 500-2000 m in the fermentation process³H, the tank pressure is 0.03-0.06 MPa, and after fermentation is carried out for 25-40 h, glutamic acid is added for continuous fermentation for 10-24 h;

(5) can placing and filter pressing

After the fermentation in the step (4) is finished, conveying the fermentation liquor to a fermentation liquor storage tank, adding 0.5-3% of filter aid into the fermentation liquor, stirring for 15-30 min, and conveying to a plate-and-frame filter press for filter pressing treatment;

(6) spray drying

Sending the filtrate subjected to filter pressing in the step (5) into a spray dryer for spray drying, wherein the air inlet temperature is controlled to be 130-180 ℃ and the air outlet temperature is controlled to be 65-95 ℃ during spraying, and the air inlet and outlet temperatures are required to be adjusted if the flowability of sprayed powder is poor and the sprayed powder is stuck to a tower;

(6) sieving and packaging

Controlling the granularity of the spray powder: all pass through an analysis sieve with the aperture of 1.19mm, more than 90 percent pass through an analysis sieve with the aperture of 0.59 mm;

packaging specification: 500 g/bag, 1000 g/bag, 10 kg/box, 15 kg/box, 20 kg/bag (barrel), 25 kg/bag (barrel), and the package is labeled according to the specification.

In addition, the whole production process is carried out under the aseptic condition, in order to avoid the pollution of the operation process, measures for preventing cross-contamination are necessary to be adopted, and pollution control measures for product cross-production specified in the production process pollution and cross-contamination management regulations are strictly executed:

1) during production, materials of different products are placed in different areas, and marks are made;

2) when the same operator feeds materials, the material feeding of one product is finished, then the material feeding of the other product is carried out, and when the operation is carried out by multiple operators, the operation is carried out on the premise that the materials of the respective products are obviously distinguished;

3) when different products are produced by using the same equipment in a cross mode, the equipment must be thoroughly cleaned according to the cleaning requirement, the natural color of the equipment can be seen after cleaning, the part directly contacted with the material has no obvious residue, and residue detection is carried out if necessary;

4) when different products are produced in a cross mode, related operation rules must be carefully read, and loss caused by process execution errors is prevented.

According to the method for producing gamma-aminobutyric acid through large-scale microbial whole-cell transformation, the adopted biotransformation method, namely the whole-cell transformation method, has the advantages of simplicity and convenience in operation, mild conditions, high raw material utilization rate, high transformation rate, low separation and purification cost and the like.

The points to be finally explained are: first, in the description of the present application, it should be noted that, unless otherwise specified and limited, the terms "mounted," "connected," and "connected" should be understood broadly, and may be a mechanical connection or an electrical connection, or a communication between two elements, and may be a direct connection, and "upper," "lower," "left," and "right" are only used to indicate a relative positional relationship, and when the absolute position of the object to be described is changed, the relative positional relationship may be changed;

secondly, in the drawings of the disclosed embodiments of the invention, only the structures related to the disclosed embodiments are referred to, and other structures can refer to common designs, and under the condition of no conflict, the same embodiment and different embodiments of the invention can be combined with each other;

finally, the above description is only for the preferred embodiment of the present invention and should not be taken as limiting the invention, and any modifications, equivalents, improvements and the like which are within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A system for producing gamma-aminobutyric acid through whole-cell transformation of microorganisms on a large scale is characterized by comprising:

an aseptic plant (100) comprising a floor one (101), a floor one (102), a floor three (103), a floor four (104), a floor five (105), and a plot area (106) disposed below the floor one (101); a first culture medium proportioning trough (107), a second culture medium proportioning trough (109) and a third culture medium proportioning trough (111) which are embedded in the ground reservoir area (106) are sequentially arranged at one end of the first floor (101);

a plurality of seed liquid culture shake flasks (200) which adopt large triangular flasks and are arranged in corresponding shaking tables on the floor five (105) for receiving the seed liquid after slant culture to carry out shake flask seed liquid culture, the bottoms of the shake flasks are connected with a hose (306) on a feeder (300) in a row continuously through a pipeline, and openings at the tops of the shake flasks are respectively connected with the first culture medium distribution groove (107) through a conveying hose and a first negative pressure suction pipe (108);

the gang-type feeder (300) is embedded on a floor slab of the fourth floor (104) and is used for receiving the seed liquid in the shake flask cultured by the seed liquid culture shake flask (200) for storage and distribution, the gang-type feeder comprises a feeding trough (301) and a base plate (318) which is laterally erected on one side of the feeding trough (301) through an extension plate (303), and a longitudinal moving mechanism used for moving the discharge valve pipe (307) to a position corresponding to the feeding trough (301) and a telescopic cylinder (327) used for controlling the opening and closing of an insertion valve plate (309) are slidably arranged on the base plate (318);

the primary seed culture cylinders (400) are embedded on a floor slab of the third floor (103) at intervals and used for receiving shake flask seed liquid distributed by the row-type feeder (300) to perform primary seed liquid culture, the tops of the primary seed culture cylinders are respectively and correspondingly connected with the discharge valve pipes (307) through pipelines, the tops of the primary seed culture cylinders are connected with the second culture medium distribution tank (109) through a second negative pressure suction pipe (110) through a conveying hose, the lower ends of the primary seed culture cylinders are located on the first floor (102), and the bottoms of the primary seed culture cylinders are respectively provided with a discharge valve; and

the fermentation units (500) are arranged on the first floor (102) and are used for receiving the primary seed culture solution cultured by the primary seed culture cylinder (400) for inoculation and fermentation; each set of fermentation units (500) comprises two tanks (521); each jar body (521) all is the corresponding arrangement rather than the top with one-level seed culture section of thick bamboo (400), and just connects through delivery hose through third negative pressure suction pipe (112) third culture medium batching groove (111) and lactic acid bacteria liquid storage tank (115) that floor four (104) were arranged through delivery hose connection respectively.

2. The system for producing gamma-aminobutyric acid through scale microbial whole cell transformation according to claim 1, wherein the feeding trough (301) is divided into independent feeding bins (302) along the length direction thereof by vertical plates (315), the bottom of each feeding bin (302) is provided with a vertically arranged discharging valve pipe (307) and a horizontally arranged valve inserting groove (308), and the valve inserting groove (308) is movably provided with a valve inserting plate (309); a plate hole (310) for communicating the discharge valve pipe (307) with the lower storage bin (302) is formed in the end face of one end of the valve inserting plate (309); the other end of the valve inserting plate (309) is connected with a compression spring (313), and the compression spring (313) is fixed on the lower side wall of the blanking groove (301) through a limiting frame (314).

3. The system for producing gamma-aminobutyric acid through scale microbial whole cell transformation according to claim 2, wherein the longitudinal moving mechanism comprises an upper sliding rail (319), a toothed plate (320), a lower sliding rail (324) and a driving motor (321) arranged on the upper sliding rail (319) through a sliding block, wherein the upper sliding rail (319) is arranged on the base plate (318) in parallel, and the driving motor comprises:

the upper sliding track (319), the tooth plate (320) and the lower sliding track (324) are arranged in parallel along the length direction of the base plate (318), and the upper sliding track (319) and the lower sliding track (324) are located at two side positions of the tooth plate (320);

the rotating shaft of the driving motor (321) is in transmission connection with a walking gear (323) through a coupler (322), and the walking gear (323) is in meshed connection with the toothed plate (320).

4. The system for producing gamma-aminobutyric acid through scale microbial whole cell transformation according to claim 3, wherein the longitudinal moving mechanism further comprises a supporting cross plate (304), a fastener (305) arranged at one end of the supporting cross plate (304), and a hose (306) fixed on the fastener (305), wherein:

one end of the supporting transverse plate (304) is connected with the sliding block on the upper sliding rail (319);

the lower end of the hose (306) is positioned at the lower storage bin (302), and the upper end of the hose is connected with the seed liquid culture shake flask (200) through the hose.

5. The system for producing gamma-aminobutyric acid through scale microbial whole cell transformation according to claim 1, wherein the fermentation unit (500) comprises a support frame (510), a cylinder mechanism (520) arranged on the support frame (510), and a turnover mechanism (530) for driving the cylinder mechanism (520) to rotate, wherein:

the supporting frame (510) comprises an electric rotating table (513), a cross beam (514) and mounting arms (515) respectively arranged at two ends of the cross beam (514), and the cross beam (514) is fixedly arranged on a rotating table top of the electric rotating table (513);

the barrel mechanism (520) comprises two tank bodies (521), a left fixing shaft (525) and a right fixing shaft (526) which are movably arranged on two side walls of the tank body (521) through bearings respectively, the tank body (521) is fixedly connected with the mounting arm (515) through the left fixing shaft (525) and the right fixing shaft (526) arranged on the two side walls of the tank body, and the tank body (521) is turned over on the mounting arm (515) by taking the left fixing shaft (525) and the right fixing shaft (526) as central axes; and

tilting mechanism (530) include upset motor (531), drive belt (532) and overlap respectively and locate belt pulley (533) on left side fixed axle (525), belt pulley (533) fixed weld in on the jar body (521), two pass through between belt pulley (533) drive belt (532) are connected, just drive belt (532) are connected upset motor (531), upset motor (531) set up in on crossbeam (514).

6. The system for large-scale production of gamma-aminobutyric acid through whole-cell transformation of microorganisms according to claim 5, wherein the support frame (510) further comprises a base (511) and a support table (512) arranged on the base (511), the two ends of the base (511) are respectively provided with a discharge through hole (516) corresponding to the lower position of the tank body (521), and the top of the support table (512) is fixedly provided with the electric rotating table (513).

7. The system for producing gamma-aminobutyric acid through scale microbial whole cell transformation according to claim 5, wherein the tank (521) is a biconical tank, the top of the tank is provided with an inoculation port (522), and the bottom of the tank is provided with a discharge port (523); an exhaust pipeline (524) is arranged at the top of the tank body (521).

8. The system for producing gamma-aminobutyric acid through whole-cell transformation of microorganisms on a large scale according to claim 5, wherein the right fixed shaft (526) is a hollow shaft, and an air inlet pipeline (527) extending into the tank body (521) is arranged in the hollow shaft.

9. The system for producing gamma-aminobutyric acid through scale microbial whole-cell transformation according to claim 1, further comprising:

the fermentation liquor storage tank (600), the plate-and-frame filter press (700), the spray dryer (800) and the packaging machine (900) are sequentially arranged on the first floor (101), and the fermentation liquor storage tank (600) is respectively communicated with the fermentation units (500) on the top of the fermentation liquor storage tank through pipelines.

10. A method for the scaled microbial whole cell transformation for the production of gamma-aminobutyric acid according to any one of claims 1 to 9, comprising the steps of:

(1) slant culture

Preparing a slant culture medium, sterilizing and cooling, and subpackaging the culture medium in a sterile test tube to prepare a test tube slant; after the test tube slant is cultured to confirm sterility, picking seed liquid and uniformly streaking in a slant culture medium, and placing the slant in an incubator for culture after streaking is finished;

controlling the culture temperature to be 30-37 ℃ and the culture time to be 8-14 h;

(2) seed liquid culture in shake flask

Preparing a seed culture medium in a first culture medium proportioning tank, inoculating the test tube slant strain cultured in the step (1) into a seed solution for culturing and shaking a flask through a first negative pressure material suction pipe under an aseptic condition after sterilization and cooling, and starting a shaking table for culturing after inoculation;

controlling the rotation speed to be 100-250 rpm and the temperature to be 30-37 ℃, and culturing for 6-12 h;

(3) first order seed culture

Weighing materials such as peptone and yeast powder according to a culture medium formula, putting the materials into a second culture medium proportioning tank, metering the volume to a metered volume, preparing a first-stage seed culture medium, sterilizing by adopting steam, and quickly cooling to 30-37 ℃ after sterilization; conveying the primary seed culture medium into a primary seed culture cylinder through a second negative pressure material suction pipe, and then distributing the shake flask seed liquid cultured in the step (2) into the corresponding primary seed culture cylinder by adopting a row type feeder;

the temperature is controlled to be 30-37 ℃, the rotating speed is 100-280 rpm, and the ventilation volume is 50-200 m in the fermentation process³H, pot pressure0.03-0.06 MPa, and no mixed bacteria in microscopic examination; sampling in the fermentation process, detecting physical and chemical indexes and sterility conditions, and culturing in a seed solution for 6-12 h;

(4) fermentation culture

Weighing materials such as peptone and yeast powder according to a culture medium formula, putting the materials into a second culture medium proportioning tank, fixing the volume to a measured volume, sterilizing by using steam, and quickly cooling to a culture temperature after the sterilization is finished; conveying the primary seed culture medium to a corresponding tank body through a third negative pressure material suction pipe, controlling the temperature, the rotating speed, the ventilation volume, the tank pressure and other control points of the fermentation tank to meet the process requirements, and then respectively inoculating the primary seed culture solution cultured in the step (3) and the lactic acid bacteria in the lactic acid bacteria liquid storage tank, wherein the addition amount of the lactic acid bacteria is 20-100 mg/L;

the temperature is controlled to be 25-37 ℃, the rotating speed is 50-130 rpm, and the ventilation volume is 500-2000 m in the fermentation process³H, the tank pressure is 0.03-0.06 MPa, and after fermentation is carried out for 25-40 h, glutamic acid is added for continuous fermentation for 10-24 h;

(5) can placing and filter pressing

After the fermentation in the step (4) is finished, conveying the fermentation liquor to a fermentation liquor storage tank, adding 0.5-3% of filter aid into the fermentation liquor, stirring for 15-30 min, and conveying to a plate-and-frame filter press for filter pressing treatment;

(6) spray drying

Sending the filtrate subjected to filter pressing in the step (5) into a spray dryer for spray drying, wherein the air inlet temperature is controlled to be 130-180 ℃ and the air outlet temperature is controlled to be 65-95 ℃ during spraying;

(6) sieving and packaging

And (5) sieving the dried powder in the step (5), and packaging and labeling by using a packaging machine according to the specification.

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