Full-automatic sterile inoculation device and method

Abstract

The invention provides a full-automatic sterile inoculation device and method, belongs to the technical field of microbial fermentation, and is used for solving the problem that in the related technology, inoculation of seed liquid is prone to bacterial contamination, in the device and method, a detachable communicating and connecting structure is arranged between a seed module for storing the seed liquid and a fermentation module for inoculating the seed liquid, and the seed liquid is inoculated into the fermentation module; reutilization of the seed liquid among a plurality of fermentation modules is realized, and external high-temperature sterilization steam is introduced by utilizing a specially designed pipeline structure to perform high-temperature sterilization on a part which is possibly contaminated, so that the contamination risk in the seed liquid inoculation process is favorably reduced.

Description

Technical Field

[0001] The present application relates to the technical field of microbial fermentation, and in particular to a fully automatic aseptic inoculation device and method. Background Art

[0002] In most industrial fermentations, whether single-strain or mixed-strain fermentations, microorganisms other than the target strain are considered foreign bacteria. Almost all fermentation industries are likely to encounter foreign bacteria contamination, which has a great impact on fermentation yield, extraction rate, product quality, etc. Therefore, preventing foreign bacteria contamination is one of the keys to ensuring the normal progress of fermentation.

[0003] Seed culture is a type of industrial fermentation, and seed culture mainly grows and reproduces bacteria. Seed liquid inoculation is one of the important links in seed culture. Seed liquid inoculation is easy to be contaminated with bacteria, and because seed liquid inoculation is at the beginning of the seed culture process, if the seed liquid is inoculated with bacteria, it is easy to bring miscellaneous bacteria into the fermentation tank and contaminate the subsequent process flow. Therefore, it is particularly important to ensure the cleanliness and sterility of seed liquid inoculation.

[0004] At present, there are two typical seed liquid inoculation methods, namely the fire ring inoculation method and the acupuncture inoculation method. Among them, the fire ring inoculation method refers to the configuration of a fire ring for sterilization at the inoculation port of the fermentation tank, and the seed liquid is quickly poured into the fermentation tank during inoculation. Although this inoculation method has the fire ring sterilization, the seed liquid will be exposed to the ambient air for a period of time and is still easy to be contaminated. In the acupuncture inoculation method, the seed liquid is stored in a specific container, one end of the pipeline is inserted into the seed liquid, and the other end is connected to the puncture needle, and a peristaltic pump is connected to the pipeline. During actual inoculation, the puncture needle package is opened, and the puncture needle is inserted into the sealant cover at the inoculation port of the fermentation tank. The peristaltic pump can be used to input the seed liquid into the fermentation tank. In this inoculation method, the puncture needle will be exposed to the air for a period of time and will contact the outer surface of the sealant cover at the inoculation port of the fermentation tank, which is still easy to be contaminated.

[0005] Therefore, it is necessary to find a method that can achieve sterile inoculation. Summary of the invention

[0006] The present application provides a fully automatic sterile inoculation device and method, which is beneficial to reducing the risk of contamination during seed liquid inoculation.

[0007] In the first aspect, the present application provides a fully automatic aseptic inoculation device. The device includes a seed liquid outlet control valve, a fermentation tank inoculation liquid control valve, and a steam control valve disposed between the seed liquid outlet control valve and the fermentation tank inoculation liquid control valve and fixedly connected to the fermentation tank inoculation liquid control valve; one end of the seed liquid outlet control valve is used to be fixedly connected to the seed module, and the other end is fixedly connected to the second condensate control valve; one end of the fermentation tank inoculation liquid control valve is used to be fixedly connected to the fermentation module, and the other end is fixedly connected to the first condensate control valve; One end of the seed liquid outlet control valve away from the seed module is detachably connected to one end of the fermentation tank inoculation liquid control valve away from the fermentation module; one end of the fermentation tank inoculation liquid control valve close to the fermentation module is detachably connected to the fermentation module; a pumping module is provided between one end of the seed liquid outlet control valve close to the seed module and the seed module; The seed module is used to store seed liquid; the pumping module is used to pump the seed liquid from the seed module to the fermentation module; and the fermentation module is used to receive the seed liquid and ferment it.

[0008] As another fully automatic aseptic inoculation device provided in the first aspect of the present application, the device comprises a seed liquid outlet control valve and a first connecting valve, one end of the seed liquid outlet control valve is used to be fixedly connected to the seed module, and the other end is fixedly connected to one end of the first connecting valve; One end of the seed liquid outlet control valve away from the seed module is also used for detachably connecting to one end of the fermentation tank inoculation liquid control valve, and the other end of the fermentation tank inoculation liquid control valve is used for fixedly connecting to the fermentation module; one end of the fermentation tank inoculation liquid control valve away from the fermentation module is also used for fixedly connecting to one end of the second connecting valve; The other end of the first connecting valve and the other end of the second connecting valve are used as a steam inlet and a condensed water outlet of the steam sterilization module respectively; A pumping module is fixedly connected between the seed liquid outlet control valve and the seed module or between the fermentation tank inoculation liquid control valve and the fermentation module; The seed module is used to store seed liquid; the pumping module is used to pump the seed liquid from the seed module to the fermentation module; and the fermentation module is used to receive the seed liquid and ferment it.

[0009] As a third fully automatic aseptic inoculation device provided in the first aspect of the present application, the device comprises a fermentation tank inoculation liquid control valve and a second connecting valve, one end of the fermentation tank inoculation liquid control valve is used to be fixedly connected to the fermentation module, and the other end is fixedly connected to one end of the second connecting valve; One end of the fermentation tank inoculation liquid control valve away from the fermentation module is also used for detachably connecting to one end of the seed liquid outlet control valve, the other end of the seed liquid outlet control valve is used for fixedly connecting to the seed module, and one end of the seed liquid outlet control valve away from the seed module is also used for fixedly connecting to one end of the first connecting valve; The other end of the first connecting valve and the other end of the second connecting valve are used as an air inlet and an outlet of the steam sterilization module respectively; A pumping module is fixedly connected between the seed liquid outlet control valve and the seed module or between the fermentation tank inoculation liquid control valve and the fermentation module; The seed module is used to store seed liquid; the pumping module is used to pump the seed liquid from the seed module to the fermentation module; and the fermentation module is used to receive the seed liquid and ferment it.

[0010] Furthermore, a diaphragm pump is fixedly connected between the seed liquid outlet control valve (3) and the seed module and / or a diaphragm pump is fixedly connected between the fermentation tank inoculation liquid control valve (6) and the fermentation module.

[0011] Furthermore, the detachable connection is achieved by a clamp joint.

[0012] Further, the pumping module is configured as a peristaltic pump.

[0013] Further, the seed module is configured as a sterile seed bottle, and the sterile seed bottle is connected to the external environment through a sterile filter.

[0014] In the second aspect, the present application provides a fully automatic aseptic inoculation method. The method uses any one of the devices described in the first aspect above; the method comprises: The seed liquid outlet control valve (3) and the fermentation tank inoculation liquid control valve (6) are initially in a closed state; Connecting the seed liquid outlet control valve (3) to the detachable connecting section of the fermentation tank inoculation liquid control valve (6); Start the steam sterilization module for a certain period of time, and use high-temperature sterilization steam to sterilize all the pipes through which the seed liquid passes during the inoculation process; The steam sterilization module is turned off, and then the pumping module is started to transport the seed liquid from the seed module to the fermentation module.

[0015] In summary, this application at least has the following beneficial effects: A fully automatic aseptic inoculation device and method are provided, which can ensure sufficient sterilization of the potentially contaminated parts of the detachable connection between the seed module and the fermentation module on the basis that the seed module and the fermentation module are detachably connected and the seed module can inoculate multiple fermentation modules, so as to reduce the risk of contamination during seed liquid inoculation.

[0016] It should be understood that the contents described in the Summary of the Invention are not intended to limit the key or important features of the embodiments of the present application, nor are they intended to limit the scope of the present application. Other features of the present application will become easily understood through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The above and other features, advantages and aspects of the embodiments of the present application will become more apparent with reference to the following detailed description in conjunction with the accompanying drawings. In the accompanying drawings, the same or similar reference numerals represent the same or similar elements, wherein: Figure 1 A schematic diagram of a fully automatic aseptic inoculation device in an embodiment of the present application is shown; In the figure, 1. seed liquid flask; 2. peristaltic pump; 3. seed liquid outlet control valve; 4. first quick-connect clamp; 5. steam control valve; 6. fermenter inoculation liquid control valve; 7. fermenter; 8. second quick-connect clamp; 9. first condensate control valve; 10. second condensate control valve. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solution and advantages of the embodiments of the present application clearer, the technical solution in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0019] In addition, the term "and / or" in this article is only a description of the association relationship between the associated objects, indicating that there can be three relationships. For example, A and / or B can represent: A exists alone, A and B exist at the same time, and B exists alone. In addition, the character " / " in this article generally indicates that the associated objects before and after are in an "or" relationship.

[0020] The present application provides a fully automatic sterile inoculation device and method, which can reduce the risk of contamination during seed liquid inoculation.

[0021] In the first aspect, the embodiment of the present application discloses a fully automatic aseptic inoculation device. The device includes a seed liquid outlet control valve 3, a fermentation tank inoculation liquid control valve 6, and a steam control valve 5 disposed between the seed liquid outlet control valve 3 and the fermentation tank inoculation liquid control valve 6 and fixedly connected to the fermentation tank inoculation liquid control valve 6; one end of the seed liquid outlet control valve 3 is used to be fixedly connected to the seed module, and the other end is fixedly connected to the second condensate control valve 10; one end of the fermentation tank inoculation liquid control valve 6 is used to be fixedly connected to the fermentation module, and the other end is fixedly connected to the first condensate control valve 10; Specifically, the end of the seed liquid outlet control valve 3 away from the seed module is detachably connected to the end of the fermentation tank inoculation liquid control valve 6 away from the fermentation module; the end of the fermentation tank inoculation liquid control valve 6 close to the fermentation module is detachably connected to the fermentation module; a pumping module is arranged between the end of the seed liquid outlet control valve 3 close to the seed module and the seed module.

[0022] The seed module is used to store seed liquid, the pumping module is used to pump the seed liquid from the seed module to the fermentation module, and the fermentation module is used to receive the seed liquid and ferment it.

[0023] Furthermore, the detachable connection is achieved by a clamp joint.

[0024] Further, the pumping module is configured as a peristaltic pump.

[0025] Further, the seed module is configured as a sterile seed bottle, and the sterile seed bottle is connected to the external environment through a sterile filter.

[0026] Figure 1 A schematic diagram of a fully automatic aseptic inoculation device in an embodiment of the present application is shown.

[0027] Reference Figure 1 As a specific example, the device includes a seed liquid flask 1, a peristaltic pump 2, a seed liquid outlet control valve 3, a first quick-connect clamp 4, a steam control valve 5, a fermenter inoculation liquid control valve 6, a second quick-connect clamp 8, and a fermenter 7, which are connected in sequence. It also includes a second condensate control valve 10 connected to the seed liquid outlet control valve 3, and a first condensate control valve 9 connected to the fermenter inoculation liquid control valve 6.

[0028] The device includes a seed preservation module, a steam sterilization module and a fermentation module, which are connected in sequence through a first quick-connect clamp 4 and a second quick-connect clamp 8, so that the seed module can be used for inoculation of the fermentation module. The seed preservation module includes a seed module (specifically a seed liquid flask 1), a peristaltic pump 2, a seed liquid outlet control valve 3 and a second condensate control valve 10; the steam sterilization module includes a steam control valve 5, a fermentation tank inoculation liquid control valve 6 and a first condensate control valve 9; and the fermentation module is specifically a fermentation tank 7.

[0029] The seed liquid flask 1 is a sterile environment. The seed liquid flask 1 can be fixedly connected to the liquid inlet of the peristaltic pump 2 through a silicone hose, and the liquid outlet of the peristaltic pump 2 can be fixedly connected to the seed liquid outlet control valve 3 through a silicone hose, and the seed liquid outlet control valve 3 is fixedly connected to the second condensed water control valve 10.

[0030] The seed liquid outlet control valve 3 is detachably connected to the fermentation tank inoculation liquid control valve 6 via a first quick-connect clamp 4 , and the fermentation tank inoculation liquid control valve 6 is detachably connected to the space inside the fermentation tank 7 via a second quick-connect clamp 4 .

[0031] The side of the fermentation tank inoculation liquid control valve 6 close to the seed liquid outlet control valve 3 is also fixedly connected to the steam control valve 5, and the side of the fermentation tank inoculation liquid control valve 6 away from the seed liquid outlet control valve 3 is also fixedly connected to the first condensed water control valve 9.

[0032] When the device is used, the seed liquid outlet control valve 3 and the fermentation tank inoculation liquid control valve 6 are normally closed, and due to the sealing effect of the seed liquid outlet control valve 3, the sterile environment of the seed liquid flask 1 can be guaranteed. When inoculation is required, the first quick-connect clamp 4 and the second quick-connect clamp 8 are used to realize the sequential connection of the above three modules, and then steam sterilization is performed. During the steam sterilization process, the steam control valve 5 is first opened, and then the first condensed water control valve 9 and the second condensed water control valve 10 are opened, and the high-temperature sterilization steam (the steam generating device for generating high-temperature sterilization steam is not shown in the figure) is used to sterilize the parts that may be contaminated with bacteria at high temperature, including the pipeline used for seed liquid inoculation in the device, for example, using 120° sterilization for 30 minutes to ensure that the parts that may be contaminated with bacteria are fully sterilized. After the sterilization is completed, the first condensed water control valve 9 and the second condensed water control valve 10 are first closed, and then the steam control valve 5 is closed. During the inoculation process, the peristaltic pump 2 and the switch connected to the inside of the fermentation tank 7 are turned on, so that the seed liquid can be safely transported to the fermentation tank 7 through the sterile environment, thereby completing the sterile inoculation process, achieving sterile inoculation, and avoiding the risk of contamination during the inoculation process.

[0033] In the aforementioned device, if the pumping efficiency of the peristaltic pump 2 is sufficient, the seed liquid outlet control valve 3 may not be configured, and only two valves are configured at the corresponding position to control opening and closing. The peristaltic pump 2 can be configured on the side close to the seed liquid flask 1 or on the side close to the fermentation tank 7. The first quick-connect clamp 4 and the second quick-connect clamp 8 can realize the detachable connection between the seed preservation module, the steam sterilization module and the fermentation module. As such, various deformations that can be thought of by those skilled in the art without creative labor based on the example structure should all fall within the scope disclosed in this application.

[0034] It should be understood that the core improvement of the device is to use a clamp joint to achieve a detachable connection between the seed liquid flask 1 and the fermentation tank 7 and a partial structure that can be connected to external sterilization steam. Therefore, the device may also only include two necessary clamp joints and a steam inlet valve and a condensate discharge valve. Of course, considering that the device is an individual that can be detachably sold and configured separately, the two parts connected by the clamp joint should also be disclosed as independent products as an embodiment of the present application.

[0035] That is, another fully automatic aseptic inoculation device provided in the first aspect of the present application. The device comprises a seed liquid outlet control valve 3 and a first connecting valve, wherein one end of the seed liquid outlet control valve 3 is used to be fixedly connected to the seed module, and the other end is fixedly connected to one end of the first connecting valve; The end of the seed liquid outlet control valve 3 away from the seed module is also used for detachable communication with one end of the fermentation tank inoculation liquid control valve 6, and the other end of the fermentation tank inoculation liquid control valve 6 is used for fixed communication with the fermentation module; the end of the fermentation tank inoculation liquid control valve 6 away from the fermentation module is also used for fixed communication with one end of the second communication valve. The other end of the first communication valve and the other end of the second communication valve are used as the steam inlet and condensed water outlet of the steam sterilization module respectively.

[0036] A pumping module is fixedly connected between the seed liquid outlet control valve 3 and the seed module or between the fermentation tank inoculation liquid control valve 6 and the fermentation module.

[0037] The seed liquid flask 1 is used to store the seed liquid, the pumping module is used to pump the seed liquid from the seed module to the fermentation module, and the fermentation module is used to receive the seed liquid.

[0038] As the third fully automatic aseptic inoculation device provided in the first aspect of the present application, the device comprises a fermentation tank inoculation liquid control valve 6 and a second connecting valve, wherein one end of the fermentation tank inoculation liquid control valve 6 is used to be fixedly connected to the fermentation module, and the other end is fixedly connected to one end of the second connecting valve.

[0039] The end of the fermentation tank inoculation liquid control valve 6 away from the fermentation module is also used for detachable communication with one end of the seed liquid outlet control valve 3, the other end of the seed liquid outlet control valve 3 is used for fixed communication with the seed module, and the end of the seed liquid outlet control valve 3 away from the seed module is also used for fixed communication with one end of the first communication valve. The other end of the first communication valve and the other end of the second communication valve are used as the air inlet and outlet of the steam sterilization module respectively.

[0040] A pumping module is fixedly connected between the seed liquid outlet control valve 3 and the seed module or between the fermentation tank inoculation liquid control valve 6 and the fermentation module.

[0041] The seed module is used to store the seed liquid; the pumping module is used to pump the seed liquid from the seed module to the fermentation module; and the fermentation module is used to receive the seed liquid and ferment it.

[0042] In a second aspect, the present application provides a fully automatic aseptic inoculation method, which is implemented using the device disclosed in the first aspect of the embodiment of the present application.

[0043] The method specifically includes: S1: The seed liquid outlet control valve 3 and the fermentation tank inoculation liquid control valve 6 are initially in the closed state; S2: Connect the seed liquid outlet control valve 3 with the detachable connecting section of the fermentation tank inoculation liquid control valve 6; S3: Start the steam sterilization module for a certain period of time, and use high-temperature sterilization steam to sterilize all the pipes through which the seed liquid passes during the inoculation process; when starting, first open the steam control valve 5, and then open the first condensate control valve 9 and the second condensate control valve 10; S4: Turn off the steam sterilization module. When closing, first close the first condensate control valve 9 and the second condensate control valve 10, and then close the steam control valve 5; start the peristaltic pump 2 to transport the seed liquid from the seed module (seed liquid flask 1) to the fermentation module (fermentation tank 7).

[0044] It should be noted that, for the aforementioned method embodiments, for the sake of simplicity of description, they are all expressed as a series of action combinations, but those skilled in the art should be aware that the present application is not limited by the described order of actions, because according to the embodiments of the present application, certain steps can be performed in other orders or simultaneously. Secondly, those skilled in the art should also be aware that the embodiments described in the specification are all optional embodiments, and the actions and modules involved are not necessarily required by the present application.

[0045] Those skilled in the art can clearly understand that, for the convenience and brevity of description, the specific working process of the described method can refer to the corresponding process in the aforementioned device embodiment, and will not be repeated here.

[0046] In summary, this application at least has the following beneficial effects: A fully automatic aseptic inoculation device and method are provided, which can ensure sufficient sterilization of the potentially contaminated parts of the detachable connection between the seed module and the fermentation module on the basis that the seed module and the fermentation module are detachably connected and the seed module can inoculate multiple fermentation modules, so as to reduce the risk of contamination during seed liquid inoculation.

[0047] The above description is only a preferred embodiment of the present application and an explanation of the technical principles used. Those skilled in the art should understand that the scope of disclosure involved in the present application is not limited to the technical solution formed by a specific combination of the above technical features, but should also cover other technical solutions formed by any combination of the above technical features or their equivalent features without departing from the aforementioned disclosed concept. For example, the above features are replaced with the technical features with similar functions disclosed in this application (but not limited to) by each other to form a technical solution.

Claims

1. A fully automatic sterile inoculation device, characterized in that: It comprises a seed liquid outlet control valve (3), a fermentation tank inoculation liquid control valve (6), and a steam control valve (5) arranged between the seed liquid outlet control valve (3) and the fermentation tank inoculation liquid control valve (6) and fixedly connected to the fermentation tank inoculation liquid control valve (6); one end of the seed liquid outlet control valve (3) is used for fixedly connecting to the seed module, and the other end is fixedly connected to the second condensed water control valve (10); one end of the fermentation tank inoculation liquid control valve (6) is used for fixedly connecting to the fermentation module, and the other end is fixedly connected to the first condensed water control valve (10); The end of the seed liquid outlet control valve (3) away from the seed module is detachably connected to the end of the fermentation tank inoculation liquid control valve (6) away from the fermentation module; the end of the fermentation tank inoculation liquid control valve (6) close to the fermentation module is detachably connected to the fermentation module; a pumping module is provided between the end of the seed liquid outlet control valve (3) close to the seed module and the seed module; The seed module is used to store seed liquid; the pumping module is used to pump the seed liquid from the seed module to the fermentation module; and the fermentation module is used to receive the seed liquid and ferment it.

2. A fully automatic sterile inoculation device, characterized in that: It comprises a seed liquid outlet control valve (3) and a first connecting valve, wherein one end of the seed liquid outlet control valve (3) is used to be fixedly connected to the seed module, and the other end is fixedly connected to one end of the first connecting valve; One end of the seed liquid outlet control valve (3) away from the seed module is also used for detachably connecting to one end of the fermentation tank inoculation liquid control valve (6), and the other end of the fermentation tank inoculation liquid control valve (6) is used for fixedly connecting to the fermentation module; one end of the fermentation tank inoculation liquid control valve (6) away from the fermentation module is also used for fixedly connecting to one end of the second connecting valve; The other end of the first connecting valve and the other end of the second connecting valve are used as a steam inlet and a condensed water outlet of the steam sterilization module respectively; A pumping module is fixedly connected between the seed liquid outlet control valve (3) and the seed module or between the fermentation tank inoculation liquid control valve (6) and the fermentation module; The seed module is used to store seed liquid; the pumping module is used to pump the seed liquid from the seed module to the fermentation module; and the fermentation module is used to receive the seed liquid and ferment it.

3. A fully automatic sterile inoculation device, characterized in that: It comprises a fermentation tank inoculation liquid control valve (6) and a second connecting valve, wherein one end of the fermentation tank inoculation liquid control valve (6) is used to be fixedly connected to the fermentation module, and the other end is fixedly connected to one end of the second connecting valve; One end of the fermentation tank inoculation liquid control valve (6) away from the fermentation module is also used for detachably connecting to one end of the seed liquid outlet control valve (3), the other end of the seed liquid outlet control valve (3) is used for fixedly connecting to the seed module, and the end of the seed liquid outlet control valve (3) away from the seed module is also used for fixedly connecting to one end of the first connecting valve; The other end of the first connecting valve and the other end of the second connecting valve are used as an air inlet and an outlet of the steam sterilization module respectively; A pumping module is fixedly connected between the seed liquid outlet control valve (3) and the seed module or between the fermentation tank inoculation liquid control valve (6) and the fermentation module; The seed module is used to store seed liquid; the pumping module is used to pump the seed liquid from the seed module to the fermentation module; and the fermentation module is used to receive the seed liquid and ferment it.

4. The device according to any one of claims 1 to 3, characterized in that: A diaphragm pump is fixedly connected between the seed liquid outlet control valve (3) and the seed module and / or a diaphragm pump is fixedly connected between the fermentation tank inoculation liquid control valve (6) and the fermentation module.

5. The device according to any one of claims 1 to 3, characterized in that: The detachable connection is achieved by a clamp joint.

6. The device according to any one of claims 1 to 3, characterized in that: The pumping module is configured as a peristaltic pump.

7. The device according to any one of claims 1 to 3, characterized in that: The seed module is configured as a sterile seed bottle, and the sterile seed bottle is connected to the external environment through a sterile filter.

8. A fully automatic aseptic inoculation method, characterized in that: Using the device as described in any one of claims 1 to 7; the method comprises: The seed liquid outlet control valve (3) and the fermentation tank inoculation liquid control valve (6) are initially in a closed state; Connecting the seed liquid outlet control valve (3) to the detachable connecting section of the fermentation tank inoculation liquid control valve (6); Start the steam sterilization module for a certain period of time, and use high-temperature sterilization steam to sterilize all the pipes through which the seed liquid passes during the inoculation process; The steam sterilization module is turned off, and then the pumping module is started to transport the seed liquid from the seed module to the fermentation module.

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