阅读说明：本技术 一种微生物菌种的扩培装置及方法 (Microbial strain expanding culture device and method ) 是由 剧永涛 张雪娜 李秀锋 张�杰 刘斌 牛文业 于 2021-07-19 设计创作，主要内容包括：本发明公开了一种微生物菌种的扩培方法,包括如下步骤：步骤100、将需氧型菌种投入培养罐内,向培养罐内通入无菌空气并对培养液进行同步搅拌；步骤200、当培养罐内的气体压强达到预设最高值后,停止无菌空气的通入与以及培养液的搅拌；步骤300、当培养罐内的气体压强低于预设最低值时,继续向培养罐内通入无菌空气并进行同步搅拌；步骤400、重复步骤200和步骤300直至培养罐内的菌种数量达到指定数量级；本发明还提供了一种微生物菌种的扩培装置；本发明将微生物培养过程中氧气的通入改为间歇式,利用高压有助于氧气的溶解,在通氧结束后,停止搅拌,创造微生物不受打扰的增殖状态,避免菌体受损,从而提高了菌群的繁殖速度与质量。(The invention discloses a method for expanding culture of microbial strains, which comprises the following steps: step 100, putting aerobic strains into a culture tank, introducing sterile air into the culture tank, and synchronously stirring culture solution; step 200, stopping introducing sterile air and stirring the culture solution when the gas pressure in the culture tank reaches a preset maximum value; step 300, when the gas pressure in the culture tank is lower than a preset minimum value, continuously introducing sterile air into the culture tank and synchronously stirring; step 400, repeating step 200 and step 300 until the number of strains in the culture tank reaches a specified magnitude; the invention also provides a device for expanding culture of microbial strains; the invention changes the introduction of oxygen in the process of culturing the microorganisms into an intermittent type, utilizes high pressure to facilitate the dissolution of the oxygen, stops stirring after the introduction of the oxygen is finished, creates a proliferation state without disturbing the microorganisms, avoids the damage of thalli, and improves the propagation speed and the quality of floras.)

1. A method for expanding culture of microbial strains is characterized by comprising the following steps:

step 100, putting aerobic strains into a culture tank, introducing sterile air into the culture tank, and synchronously stirring culture solution;

step 200, stopping introducing sterile air and stirring the culture solution when the gas pressure in the culture tank reaches a preset maximum value;

step 300, when the gas pressure in the culture tank is lower than a preset minimum value, continuously introducing sterile air into the culture tank and synchronously stirring;

step 400, repeating step 200 and step 300 until the number of strains in the culture tank reaches the specified magnitude.

2. The method for expanding culture of microbial species according to claim 1, further comprising, before introducing the sterile air in step 100: and establishing an independent cavity in the culture tank so that the cavity comprises a gap between the tank body and the tank cover in the culture tank, and keeping the air pressure in the cavity to be more than 1 standard atmospheric pressure so as to prevent external air from entering the interior of the culture tank along the gap between the tank body and the tank cover.

3. An apparatus for use in a method for the expanded culture of a microbial species according to any one of claims 1 to 2, comprising:

a culture tank (10) for providing an expanded culture space for microbial strains;

an aeration mechanism (20) in communication with the interior cavity of the culture tank (10), the aeration mechanism (20) for providing sterile air to the culture tank (10);

the air pressure feedback mechanism (30) is arranged inside the culture tank (10), and the air pressure feedback mechanism (30) is used for driving the ventilation mechanism (20) to stop ventilation to the culture tank (10) after the air pressure in the culture tank (10) reaches a preset value;

the pneumatic stirring mechanism (40) is arranged in the culture tank (10), and the pneumatic stirring mechanism (40) takes the sterile air introduced by the ventilation mechanism (20) as an air source to complete self-stirring action.

4. The apparatus for expanding culture of a microorganism strain according to claim 3, wherein said predetermined value comprises a predetermined lowest value and a predetermined highest value;

when the air pressure in the culture tank reaches a preset maximum value, the air pressure feedback mechanism drives the ventilation mechanism (20) to stop ventilation to the culture tank (10);

when the air pressure in the culture tank is lower than a preset minimum value, the air pressure feedback mechanism drives the ventilation mechanism to ventilate the culture tank.

5. The device for expanding a microbial species according to claim 4, wherein said culture tank (10) comprises a tank body (11) and a tank cover (12) covering said tank body (11), and the interior of said tank body (11) is partitioned into an upper protection chamber (14) and a lower proliferation chamber (15) by a partition (13);

when the air pressure in the culture tank (10) reaches a preset maximum value, the air pressure feedback mechanism (30) drives the ventilation mechanism (20) to ventilate the protection cavity (14);

when the air pressure in the culture tank (10) is lower than a preset minimum value, the air pressure feedback mechanism (30) drives the ventilation mechanism (20) to stop ventilation to the protection cavity (14).

6. The microbial strain propagation device according to claim 5, wherein the ventilation mechanism (20) comprises a gas source tank (21) and a working pump (22) connected to the gas source tank (21), a ventilation pipe (23) is connected to an air outlet of the working pump (22), and the air pressure feedback mechanism (30) is used for driving the ventilation pipe (23) to communicate with the protection chamber (14) or the propagation chamber (15);

an air outlet (16) is formed in the outer wall of the protection cavity (14), a pressure release valve (17) is arranged at the air outlet (16), and the pressure release valve (17) is used for conveying gas in the protection cavity (14) to the gas source tank (21) after the air pressure in the protection cavity (14) reaches a set value.

7. An apparatus for the spread culture of a microbial species according to claim 6, wherein said air pressure feedback mechanism (30) comprises a first channel (31), a second channel (32) and a power member;

the first channel (31) comprises a first opening formed on the outer peripheral surface of the partition (13), and the first opening extends along the inside of the partition (13) until penetrating through one surface of the partition (13) close to the protection cavity (14) so that the first opening is communicated with the protection cavity (14);

the first channel (32) comprises a second opening formed in the outer peripheral surface of the partition (13), the second opening and the first opening are positioned at different height positions of the same vertical line, and the second opening extends along the inside of the partition (13) until penetrating through one surface of the partition (13) close to the proliferation cavity (15) so that the second opening is communicated with the proliferation cavity (15);

an air vent (18) communicated with the air pipe (23) is formed in the outer wall of the tank body (12), and the power part is used for driving the partition part (13) to move along the inner wall of the tank body (12) so that the air vent (18) is connected with the first opening or the second opening.

8. An apparatus for the spread cultivation of a microbial species according to claim 7, wherein said spacer (13) is a piston body adherently mounted inside said culture tank (10);

the position of the first opening is lower than the height of the second opening, the power part is the gas pressure difference of the protection cavity (14) and the proliferation cavity (15), and the piston body moves to the side with smaller gas pressure intensity in the protection cavity (14) and the proliferation cavity (15) under the action of the pressure difference.

9. The microbial strain propagation device according to claim 8, wherein the pneumatic stirring mechanism (40) comprises a gas receiving pipe (41) and a gas collecting pipe (42) which are arranged in the tank body (10), one end of the gas receiving pipe (41) is connected with the gas outlet of the second channel (31), the gas collecting pipe (42) is connected with the other end of the gas receiving pipe (41), and a plurality of gas nozzles (43) are respectively arranged on the outer peripheral surface and the end surface of the gas collecting pipe (42).

10. The microbial strain propagation device according to claim 9, wherein a section of threaded extension tube (44) is arranged in the middle of the air receiving tube (41), and the bottom of the air collecting tube (42) is connected with the bottom of the tank body (12) through a spring (45) so that the air collecting tube (42) can shake up and down under the action of the air flow of the air receiving tube (41).

Technical Field

The invention relates to the technical field of microbial culture, in particular to a microbial strain expanding culture device and method.

Background

In the field of microorganisms, the culture and proliferation of strains are often involved, strains are proliferated in a proper culture medium under a certain temperature condition to expand the number, and the use requirements of the strains in the fields of human medicine, veterinary medicine, feed, cultivation and the like are met. Especially for bacillus, such as bacillus subtilis, bacillus licheniformis, bacillus cereus, clostridium butyricum and the like, fermentation and multiplication are needed in a fermentation tank, so that the number of live bacteria is obviously increased. The secondary amplification of the microorganism is to obtain pure strains, and the bacteria of each colony are divided and propagated by the same bacteria. Their genetic material is essentially the same.

For the expanding culture of aerobic bacteria, oxygen needs to be introduced into a culture medium in the culture process, meanwhile, in order to improve the dissolved amount of the oxygen, a vertical fixed tank is generally adopted in the prior art, the contact area of the oxygen and a liquid culture medium is increased by utilizing mechanical stirring so as to improve the dissolved amount of the oxygen, but thalli are more easily damaged under the shearing action generated under long-time mechanical stirring, the energy consumption is higher, generally, the power of an expanding culture tank of about 2 cubic meters is about 5kw, so that the cost control is extremely unfavorable, and enterprises lose market competitiveness.

Disclosure of Invention

The invention aims to provide a microbial strain propagation device and a method, which aim to solve the technical problems that the bacteria are easily damaged by increasing dissolved oxygen through mechanical stirring and the energy consumption is high in the prior art.

In order to solve the technical problems, the invention specifically provides the following technical scheme:

a method for expanding culture of microbial strains comprises the following steps:

step 100, putting aerobic strains into a culture tank, introducing sterile air into the culture tank, and synchronously stirring culture solution;

step 200, stopping introducing sterile air and stirring the culture solution when the gas pressure in the culture tank reaches a preset maximum value;

step 300, when the gas pressure in the culture tank is lower than a preset minimum value, continuously introducing sterile air into the culture tank and synchronously stirring;

step 400, repeating step 200 and step 300 until the number of strains in the culture tank reaches the specified magnitude.

As a preferred embodiment of the present invention, before introducing the sterile air, the method further comprises, in step 100: and establishing an independent cavity in the culture tank so that the cavity comprises a gap between the tank body and the tank cover in the culture tank, and keeping the air pressure in the cavity to be more than 1 standard atmospheric pressure so as to prevent external air from entering the interior of the culture tank along the gap between the tank body and the tank cover.

The invention also provides a device for the microbial strain propagation method, which comprises the following steps:

the culture tank is used for providing an expanding culture space of microbial strains;

the aeration mechanism is communicated with the inner cavity of the culture tank and is used for providing sterile air for the culture tank;

the air pressure feedback mechanism is arranged inside the culture tank and used for driving the ventilation mechanism to stop ventilation to the culture tank after the air pressure in the culture tank reaches a preset value;

and the pneumatic stirring mechanism is arranged in the culture tank, and the pneumatic stirring mechanism takes the sterile air introduced by the ventilation mechanism as an air source to complete self stirring action.

As a preferred scheme of the present invention, the preset value includes a preset lowest value and a preset highest value;

when the air pressure in the culture tank reaches a preset maximum value, the air pressure feedback mechanism drives the ventilation mechanism to stop ventilation in the culture tank;

when the air pressure in the culture tank is lower than a preset minimum value, the air pressure feedback mechanism drives the ventilation mechanism to ventilate the culture tank.

In a preferred embodiment of the present invention, the culture tank includes a tank body and a tank cover covering the tank body, and the tank body has an interior partitioned by a partition into an upper protection chamber and a lower proliferation chamber;

when the air pressure in the culture tank reaches a preset maximum value, the air pressure feedback mechanism drives the ventilation mechanism to ventilate the protection cavity;

when the air pressure in the culture tank is lower than a preset minimum value, the air pressure feedback mechanism drives the ventilation mechanism to stop ventilation to the protection cavity.

As a preferable scheme of the present invention, the ventilation mechanism includes a gas source tank and a working pump connected to the gas source tank, an air outlet of the working pump is connected to a ventilation pipe, and the air pressure feedback mechanism is used for driving the ventilation pipe to communicate with the protection cavity or the proliferation cavity;

the outer wall of the protection cavity is provided with an exhaust port, a pressure relief valve is arranged at the exhaust port, and the pressure relief valve is used for conveying gas in the protection cavity to the gas source tank after the gas pressure in the protection cavity reaches a set value.

As a preferable scheme of the invention, the air pressure feedback mechanism comprises a first channel, a second channel and a power piece;

the first channel comprises a first opening formed in the peripheral surface of the isolating piece, and the first opening extends along the inside of the isolating piece until penetrating through one surface of the isolating piece, which is close to the protection cavity, so that the first opening is communicated with the protection cavity;

the first channel comprises a second opening formed in the outer peripheral surface of the partition, the second opening and the first opening are located at different height positions of the same vertical line, and the second opening extends along the inside of the partition until penetrating through one surface of the partition, which is close to the proliferation cavity, so that the second opening is communicated with the proliferation cavity;

the outer wall of the tank body is provided with a vent communicated with the vent pipe, and the power part is used for driving the isolating part to move along the inner wall of the tank body so that the vent is connected with the first opening or the second opening.

As a preferred scheme of the invention, the isolating piece is a piston body which is arranged inside the culture tank in an adherent way;

the position of the first opening is lower than the height of the second opening, the power part is the gas pressure difference between the protection cavity and the proliferation cavity, and the piston body moves to the side with smaller gas pressure in the protection cavity and the proliferation cavity under the action of the pressure difference.

As a preferable scheme of the present invention, the pneumatic stirring mechanism includes an air receiving pipe and an air collecting pipe disposed in the tank body, one end of the air receiving pipe is connected to the air outlet of the second channel, the air collecting pipe is connected to the other end of the air receiving pipe, and a plurality of air nozzles are disposed on the outer circumferential surface and the end surface of the air collecting pipe.

As a preferred scheme of the invention, a section of threaded telescopic pipe is arranged in the middle of the air receiving pipe, and the bottom of the air collecting pipe is connected with the bottom of the tank body through a spring so that the air collecting pipe can shake up and down under the action of the airflow of the air receiving pipe.

Compared with the prior art, the invention has the following beneficial effects:

the invention changes the introduction of oxygen in the microbial culture process into an intermittent type, firstly, the oxygen introduction and the stirring are simultaneously started, the gas pressure in the culture tank is continuously increased in the oxygen introduction process, the high pressure is favorable for dissolving the oxygen, the stirring is matched to ensure that the dissolution rate of the oxygen is better and the dissolved oxygen in the culture is uniformly distributed, after the oxygen introduction is finished, the stirring is stopped, the proliferation state of the microbes without disturbance is created, the thalli are prevented from being damaged, when the oxygen in the culture tank consumes a part of the oxygen in the tank, the oxygen is continuously introduced into the culture tank, so that the oxygen quantity in the culture tank is ensured to be sufficient, and the propagation speed and the quality of florae are improved.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.

FIG. 1 is a schematic structural diagram of an expanding culture device according to an embodiment of the present invention;

fig. 2 is a flowchart of an extended culture method according to an embodiment of the present invention.

The reference numerals in the drawings denote the following, respectively:

10-culture tank; 20-a venting mechanism; 30-a pneumatic pressure feedback mechanism; 40-a pneumatic stirring mechanism;

11-a tank body; 12-can lid; 13-a spacer; 14-a protection cavity; 15-proliferation chamber; 16-an exhaust port; 17-a pressure relief valve; 18-a vent;

21-a gas source tank; 22-a working pump; 23-a breather pipe;

31-a first channel; 32-a second channel;

41-air receiving pipe; 42-a gas collecting pipe; 43-air jet; 44-threaded telescoping tubes; 45-spring.

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.

As shown in FIG. 1, the present invention provides a method for expanding and culturing microbial strains, comprising the following steps:

step 100, putting aerobic strains into a culture tank, introducing sterile air into the culture tank, and synchronously stirring culture solution, wherein in the aeration process, the gas pressure in the culture tank is increased to be beneficial to dissolving oxygen, and meanwhile, the contact area of the oxygen and the culture solution is increased by matching with stirring operation, so that the dissolved oxygen is further increased, and the dissolved oxygen is uniformly distributed to all parts of the culture solution;

step 200, after the gas pressure in the culture tank reaches a preset maximum value, stopping introducing sterile air to avoid the overload operation of the culture tank caused by the overlarge gas pressure in the culture tank, and simultaneously stopping stirring of a culture solution to create a state that aerobic bacteria are propagated and are not disturbed by mechanical stirring;

step 300, when oxygen in the culture tank is partially consumed by the oxygen demand in the culture tank, and the gas pressure in the culture tank is lower than a preset minimum value, continuously introducing sterile air into the culture tank and synchronously stirring;

step 400, repeating step 200 and step 300 until the number of strains in the culture tank reaches the specified magnitude.

Wherein, in step 100, before introducing the sterile air, the method further comprises: establishing an independent cavity in the culture tank so that the cavity comprises a gap between the tank body and the tank cover in the culture tank, and keeping the gas pressure in the cavity to be more than 1 standard atmospheric pressure.

The purpose of the air pressure in the cavity being greater than 1 standard atmospheric pressure is to make the pressure in the cavity greater than the pressure outside the tank body, so that when a small air leakage gap occurs between the tank body and the tank cover, outside air cannot enter along the gap between the tank body and the tank cover and pollute the inside of the culture tank.

As shown in FIG. 2, the present invention also provides an apparatus for use in the method for expanding a microorganism strain, comprising:

a culture tank 10 for providing an expanded culture space for the microbial strains;

an aeration mechanism 20 communicating with the inner cavity of the culture tank 10, the aeration mechanism 20 being configured to provide sterile air to the culture tank 10;

the air pressure feedback mechanism 30 is arranged inside the culture tank 10, and the air pressure feedback mechanism 30 is used for driving the ventilation mechanism 20 to stop ventilation to the culture tank 10 after the air pressure in the culture tank 10 reaches a preset value;

and the pneumatic stirring mechanism 40 is arranged in the culture tank 10, and the pneumatic stirring mechanism 40 takes the sterile air introduced by the ventilation mechanism 20 as an air source to complete self-stirring action.

The preset value comprises a preset lowest value and a preset highest value;

when the air pressure in the culture tank reaches a preset maximum value, the air pressure feedback mechanism drives the ventilation mechanism 20 to stop ventilation to the culture tank 10;

when the air pressure in the culture tank is lower than a preset minimum value, the air pressure feedback mechanism drives the ventilation mechanism to ventilate the culture tank.

Specifically, the culture tank 10 includes a tank body 11 and a tank cover 12 covering the tank body 11, and the inside of the tank body 11 is partitioned into an upper protection chamber 14 and a lower proliferation chamber 15 by a partition 13;

keeping the gas pressure in the protection cavity 14 to be greater than 1 standard atmospheric pressure can prevent external air from entering the tank body 12 along the gap between the tank body 12 and the tank cover 11, wherein there are various ways to increase the pressure, in the embodiment of the present invention, in order to simplify the device structure and avoid introducing a new pressurization structure, preferably, when the gas pressure in the culture tank 10 reaches a preset maximum value, the gas pressure feedback mechanism 30 drives the ventilation mechanism 20 to ventilate the protection cavity 14;

when the air pressure in the culture tank 10 is lower than a preset minimum value, the air pressure feedback mechanism 30 drives the ventilation mechanism 20 to stop ventilation to the protection cavity 14.

The ventilation mechanism 20 comprises a gas source tank 21 and a working pump 22 connected with the gas source tank 21, a ventilation pipe 23 is connected with an air outlet of the working pump 22, and the air pressure feedback mechanism 30 is used for driving the ventilation pipe 23 to be communicated with the protection cavity 14 or the proliferation cavity 15;

in order to avoid the overlarge pressure intensity of the gas in the protection cavity 14, an exhaust port 16 is formed in the outer wall of the protection cavity 14, a pressure relief valve 17 is arranged at the exhaust port 16, and the pressure relief valve 17 is used for conveying the gas in the protection cavity 14 into the gas source tank 21 after the gas pressure in the protection cavity 14 reaches a set value.

In the present embodiment, the pneumatic feedback mechanism 30 includes a first channel 31, a second channel 32, and a power member;

the first channel 31 includes a first opening formed on the outer peripheral surface of the partition 13, and the first opening extends along the inside of the partition 13 until penetrating through a surface of the partition 13 close to the protection cavity 14 so that the first opening is communicated with the protection cavity 14;

the first channel 32 comprises a second opening formed on the outer peripheral surface of the partition 13, the second opening and the first opening are located at different height positions of the same vertical line, and the second opening extends along the inside of the partition 13 until penetrating through one surface of the partition 13 close to the proliferation cavity 15 so that the second opening is communicated with the proliferation cavity 15;

an air vent 18 communicated with the air pipe 23 is formed in the outer wall of the tank body 12, and the power part is used for driving the partition part 13 to move along the inner wall of the tank body 12 so that the air vent 18 is connected with the first opening or the second opening to realize the gas introduction of the protection cavity and the proliferation cavity.

The power member may be a telescopic rod or the like arranged inside the tank body 12 to realize the movement of the partition 13, and in the embodiment of the present invention, preferably, the partition 13 is a piston body which is installed inside the culture tank 10 in an adherent manner;

the position of the first opening is lower than the height of the second opening, the power part is the gas pressure difference between the protection chamber 14 and the proliferation chamber 15, and the piston body moves to the side with smaller gas pressure intensity in the protection chamber 14 and the proliferation chamber 15 under the action of the pressure difference.

When the second opening is connected with the vent 18, sterile air is introduced into the proliferation cavity 15, when the gas pressure in the proliferation cavity 15 is greater than that in the protection cavity 14, the piston body moves upwards until the second opening is not communicated with the vent 18, at the moment, the first opening positioned below the second opening is communicated with the vent, the piston body continues to move upwards until the gas pressure in the protection cavity 14 is greater than that in the proliferation cavity 15, the proliferation cavity 15 continues to be ventilated, and the reciprocating circulation is carried out until strains proliferate to the preset magnitude.

In the embodiment of the present invention, the stirring mode of the culture solution is preferably a pneumatic stirring mode, and compared with a mechanical stirring mode, the destructive power of the culture solution on the bacteria is smaller, and meanwhile, the pneumatic stirring mode can use sterile air as an air source to realize automatic and synchronous stirring, specifically, the pneumatic stirring mechanism 40 includes an air receiving pipe 41 and an air collecting pipe 42 which are arranged in the tank 10, one end of the air receiving pipe 41 is connected with the air outlet of the second channel 31, the air collecting pipe 42 is connected to the other end of the air receiving pipe 41, a plurality of air nozzles 43 are respectively arranged on the outer circumferential surface and the end surface of the air collecting pipe 42, and multi-directional stirring of the culture solution is realized by using the air nozzles 43 in a plurality of directions.

In addition, a section of threaded extension tube 44 is arranged in the middle of the air receiving tube 41, the bottom of the air collecting tube 42 is connected with the bottom of the tank body 12 through a spring 45, so that the air collecting tube 42 can vertically shake under the action of the airflow of the air receiving tube 41, the radiation range of the air jet 43 is wider during shaking, and the stirring efficiency is further improved.

The above embodiments are only exemplary embodiments of the present application, and are not intended to limit the present application, and the protection scope of the present application is defined by the claims. Various modifications and equivalents may be made by those skilled in the art within the spirit and scope of the present application and such modifications and equivalents should also be considered to be within the scope of the present application.