Bacillus subtilis fermentation device

Abstract

The utility model discloses a bacillus subtilis fermentation device which comprises a fermentation tank I, a primary seed tank, a secondary seed tank, a fermentation tank II and a liquid storage tank which are connected in sequence, the fermentation tank I is provided with a liquid inlet connected with hydrochloric acid and ammonia water pipelines, and the lower end of the fermentation tank I is provided with a liquid outlet connected with the primary seed tank; the first-stage seed tank is provided with a liquid inlet connected with liquid alkali, ammonia water and a dilute acid pipeline, the second-stage seed tank is provided with a liquid inlet connected with dilute alkali, ammonia water and a dilute acid pipeline, the fermentation tank is provided with a liquid inlet connected with dilute alkali, ammonia water and a dilute acid pipeline, the fermentation tank is further provided with a feeding pipeline connected with a fructose pipeline and a culture solution distribution station, and a liquid outlet of the fermentation tank II is connected with a liquid storage tank. The psicose fermentation device is reasonable in structural design, the primary seed tank and the secondary seed tank are mainly used for enabling spores to germinate, grow and breed into thalli, and the thalli can grow quickly after being inoculated into the fermentation tank II to reach a certain amount of thalli, so that the synthesis of products is facilitated, and the yield of psicose is increased.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the technical field of biological fermentation equipment, and specifically relates to a bacillus subtilis fermentation device. BACKGROUND

[0002] As a very low content six-carbon sugar in nature, allulose is a D-fructose C-3 epimer. D-allulose is difficult to be digested and absorbed, and almost does not provide energy for life activities, and thus is a very useful low-calorie sweetener. In the field of medicine and health, D-allulose can inhibit fat liver enzymes and intestinal ɑ-glycosidase, thereby reducing the accumulation of fat in the body and inhibiting the rise of blood glucose concentration. In the field of food application, allulose has the advantages of high sweetness, good solubility, low calorie and low glycemic response, and is considered to be one of the most ideal sucrose substitutes. However, the preparation method of allulose is generally enzyme immobilized conversion, and the yield is limited. As D-allulose-3-epimerase is a necessary enzyme for converting F97 syrup into allulose, therefore, it is crucial to improve the yield of D-allulose-3-epimerase, and therefore, the company has developed a process for preparing allulose by using bacillus subtilis fermentation broth. In the production process, the bacillus subtilis after culture needs to be fermented. SUMMARY

[0003] The utility model aims at providing a bacillus subtilis fermentation device to effectively obtain bacillus subtilis fermentation broth meeting the process conditions, and thereby improve the preparation yield of allulose.

[0004] To achieve the above-mentioned purposes, the utility model adopts the technical scheme of:

[0005] A bacillus subtilis fermentation device, which comprises fermentation tanks I, a primary seed tank, a secondary seed tank, fermentation tanks II and a liquid storage tank connected in sequence,

[0006] The fermentation tank I is provided with a liquid inlet connected with hydrochloric acid and ammonia water pipelines, and a liquid outlet at the lower end connected with the primary seed tank;

[0007] The primary seed tank is provided with a liquid inlet connected with liquid alkali, ammonia water and dilute acid pipelines, and a liquid outlet at the lower end connected with the secondary seed tank;

[0008] The secondary seed tank is provided with a liquid inlet connected with dilute alkali, ammonia water and dilute acid pipelines, and a liquid outlet at the lower end connected with the fermentation tank;

[0009] The fermentation tank is provided with a liquid inlet connected with dilute alkali, ammonia water and dilute acid pipelines, and a feed pipeline connected with a fructose pipeline and a culture liquid distribution station, and the liquid outlet of the fermentation tank II is connected with the liquid storage tank.

[0010] The fermenter I, the first-stage seed tank, the second-stage seed tank and the fermenter II are provided with air inlets and air outlets, the air inlets are communicated with purified air inlet pipelines, and the purified air can provide dissolved oxygen required for growth and metabolism of the bacterial strains and maintain the DO value.

[0011] Further, in order to maintain the temperature of the tank body and make the bacterial strains grow and reproduce at a suitable temperature, the fermenter I, the first-stage seed tank, the second-stage seed tank and the fermenter II are heat preservation tanks, the heat preservation cavities of the tank bodies are provided with liquid inlets connected with constant-temperature water pipelines, circulating water pipelines and steam pipelines; the first-stage seed tank and the second-stage seed tank are provided with feed inlets connected with the base material distribution station.

[0012] Further, the fermenter I is provided with a dosing device for adding antibiotics, defoaming agents and the like; and the fermenter I is provided with a stirring mechanism.

[0013] Further, the device further comprises a PLC controller, the fermenter I, the first-stage seed tank, the second-stage seed tank and the fermenter II are provided with pH meters connected with the PLC controller, each liquid pipeline is provided with an electric control valve connected with the PLC controller, and the PLC controller is used for controlling the opening and closing of the valve.

[0014] The working process of the device is as follows: after initial culture of the bacterial strains, high-density fermentation culture is carried out in the fermenter I; after the high-density fermentation culture, the Bacillus subtilis is cultured in the first-stage seed tank; after the Bacillus subtilis grows to a certain scale, the Bacillus subtilis is further cultured in the second-stage seed tank; finally, the Bacillus subtilis in the second-stage seed tank is transferred into the fermenter II for fermentation to produce D-allulose-3-epimerase, the whole process flow can be controlled by the PLC control device to realize automation and continuous production of D-allulose-3-epimerase.

[0015] The device has the advantages that: the device has a reasonable structure, the first-stage seed tank and the second-stage seed tank are mainly used for spore germination, growth and reproduction into mycelium (filament) bodies, and the mycelium bodies can grow rapidly in the fermenter II to reach a certain amount of mycelium bodies, thereby facilitating product synthesis; after passing through the first-stage seed tank and the second-stage seed tank, the growing and reproducing mycelium bodies can grow faster in the fermenter, which is beneficial to synthesis of D-allulose-3-epimerase and improves the yield of allulose. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 The device is a process flow block diagram.

[0017] 1-fermenter I, 2-first-stage seed tank, 3-second-stage seed tank, 4-fermenter II, 5-liquid storage tank, 6-hydrochloric acid pipeline, 7-ammonia pipeline, 8-diluted acid pipeline, 9-liquid alkali pipeline, 10-diluted alkali pipeline, 11-base material distribution station, 12-fructose pipeline, 13-nutrient liquid distribution station. DETAILED DESCRIPTION

[0018] In order to make the above objectives, features and advantages of the present application more apparent, specific embodiments of the present application will be described in detail below with reference to the accompanying drawings.

[0019] In the following description, a large number of specific details are set forth in order to provide a thorough understanding of the present application, but the present application can also be implemented in other ways different from those described herein, and those skilled in the art can make similar generalizations without departing from the connotation of the present application, therefore the present application is not limited by the specific embodiments disclosed below.

[0020] A bacillus subtilis fermentation device, which comprises fermentation tank I, first-stage seed tank, second-stage seed tank, fermentation tank II and liquid storage tank connected in sequence,

[0021] The fermentation tank I is provided with a liquid inlet connected with hydrochloric acid and ammonia water pipeline, and a liquid outlet at the lower end is connected with the first-stage seed tank;

[0022] The first-stage seed tank is provided with a liquid inlet connected with liquid alkali, ammonia water and dilute acid pipeline, and a liquid outlet at the lower end is connected with the second-stage seed tank;

[0023] The second-stage seed tank is provided with a liquid inlet connected with dilute alkali, ammonia water and dilute acid pipeline, and a liquid outlet at the lower end is connected with the fermentation tank;

[0024] The fermentation tank is provided with a liquid inlet connected with dilute alkali, ammonia water and dilute acid pipeline, and the fermentation tank is also provided with a feed pipeline connected with fructose pipeline and culture liquid distribution station, and the liquid outlet of the fermentation tank II is connected with the liquid storage tank.

[0025] The fermentation tank I, the first-stage seed tank, the second-stage seed tank and the fermentation tank II are provided with gas inlets and gas outlets, and the gas inlets are connected with purified air inlet pipeline.

[0026] Further, in order to maintain the temperature of the tank body and make the bacteria grow and reproduce at a suitable temperature, the fermentation tank I, the first-stage seed tank, the second-stage seed tank and the fermentation tank II are heat preservation tanks, the tank body is a double-wall tank structure, the double walls are heat preservation cavities, the heat preservation cavities are provided with liquid inlets and liquid return inlets connected with constant temperature water and steam pipeline, and the first-stage seed tank and the second-stage seed tank are provided with feed inlets connected with bottom material distribution station.

[0027] Further, the fermentation tank I is provided with a dosing device for adding antibiotics, defoaming agent and the like, and the fermentation tank I is provided with a stirring mechanism.

[0028] Further, the device further comprises a PLC controller, the fermentation tank I, the first-stage seed tank, the second-stage seed tank and the fermentation tank II are provided with pH meters connected with the PLC controller, each liquid pipeline is provided with an electric control valve, and the PLC controller is connected with the electric control valve to control the opening and closing of the valve.

[0029] The working process of this invention is as follows: After the initial culture of the bacterial strain, high-density fermentation culture is carried out in fermenter I. After high-density fermentation culture, the strain is transferred to a primary seed tank for further culture of Bacillus subtilis. After the Bacillus subtilis grows to a certain scale, it is transferred to a secondary seed tank for further culture. Finally, the Bacillus subtilis in the secondary seed tank is transferred to fermenter II for fermentation to produce D-allulose-3-epimerase. The entire process can be controlled by a PLC control device to achieve automation and continuous production of D-allulose-3-epimerase.

[0030] The primary and secondary seed tanks primarily function to germinate spores, allowing them to grow and multiply into mycelium. This mycelium, when introduced into fermenter II, grows rapidly, reaching a sufficient cell volume to facilitate product synthesis. After passing through the primary and secondary seed tanks, the mycelium can grow even faster in the fermenter, promoting the synthesis of D-allulose isomerase, which is beneficial for the cultured Bacillus subtilis. Bacillus subtilis The fermentation broth obtained from OSYZTT-01 fermentation can be used to convert fructose and allulose, and the conversion rate can reach more than 30%, which greatly improves the yield of allulose.

Claims

1. A Bacillus subtilis fermentation device, characterized in that: It comprises fermenter I, primary seed tank, secondary seed tank, fermenter II, and storage tank connected in sequence. Fermentation tank I is equipped with an inlet for connecting hydrochloric acid and ammonia water pipelines, and its lower end is equipped with an outlet for connecting to the primary seed tank; The primary seed tank is equipped with an inlet connected to liquid alkali, ammonia, and dilute acid pipelines, and its lower end is equipped with an outlet connected to the secondary seed tank. The secondary seed tank is equipped with an inlet for connecting to dilute alkali, ammonia, and dilute acid pipelines, and an outlet at its lower end for connecting to the fermentation tank. The fermenter is equipped with an inlet for connecting to dilute alkali, ammonia, and dilute acid pipelines. The fermenter is also equipped with a feed pipeline for connecting to fructose pipelines and a culture medium distribution station. The outlet of fermenter II is connected to a storage tank.

2. The Bacillus subtilis fermentation apparatus according to claim 1, characterized in that: The fermentation tank I, primary seed tank, secondary seed tank, and fermentation tank II are equipped with air inlets and exhaust outlets, with the air inlets connected to a purified air intake pipeline.

3. The Bacillus subtilis fermentation apparatus according to claim 1, characterized in that: Fermentation tank I, primary seed tank, secondary seed tank, and fermentation tank II are all insulated tanks.

4. The Bacillus subtilis fermentation apparatus according to claim 3, characterized in that: The primary and secondary seed tanks are equipped with inlets that connect to the bottom material distribution station.

5. The Bacillus subtilis fermentation apparatus according to claim 1, characterized in that: Fermentation tank I is equipped with a dosing device; fermentation tank I also has its own stirring mechanism.

6. The Bacillus subtilis fermentation apparatus according to claim 1, characterized in that: The device also includes a PLC controller. Fermentation tank I, primary seed tank, secondary seed tank, and fermentation tank II are equipped with pH meters connected to the PLC controller.

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