阅读说明：本技术 一种用于酒精分解的芽孢菌选育方法 (Method for breeding spore bacteria for alcohol decomposition ) 是由 韦柯柯 李胜 高文功 黄时海 于 2021-08-23 设计创作，主要内容包括：本发明公开了一种用于酒精分解的芽孢菌选育方法,方法包括以下步骤：S1、芽孢菌的液体菌种接种至选育装置内的培养基进行培育,接种量为6-10％；培养基包括以下重量份原料：玉米粉30-50g/L,葡萄糖5-8g/L,蛋白胨5-10g/L,龙爪果粉20-40g/L,葛根粉20-30g/L,硫酸镁0.1-0.2g/L,磷酸二氢钠0.3-0.8g/L,亚硫酸钠0.1-0.3g/L；S2、步骤S1中的芽孢菌的液体菌培养24-48h,温度为35-40℃,然后培养液加入到酒精样品中,进行解酒能力筛选；S3、步骤S2中的酒精样品经离心,取上层溶液,然后再次重复根据S1和S2的步骤进行操作,至选育出适合的芽孢菌。本发明通过先培育芽孢菌,再进行解酒能力筛选,逐步选育出具有较高解酒能力的菌种,操作高效快捷；使用的选育装置能在相同环境下进行不同酒精度的分解对比筛选,提高选育效率。(The invention discloses a method for breeding spore bacteria for alcohol decomposition, which comprises the following steps: s1, inoculating the liquid strains of the spore bacteria to a culture medium in a breeding device for cultivation, wherein the inoculation amount is 6-10%; the culture medium comprises the following raw materials in parts by weight: 30-50g/L corn flour, 5-8g/L glucose, 5-10g/L peptone, 20-40g/L dragon claw fruit powder, 20-30g/L kudzu root powder, 0.1-0.2g/L magnesium sulfate, 0.3-0.8g/L sodium dihydrogen phosphate and 0.1-0.3g/L sodium sulfite; s2, culturing the liquid bacteria of the spore bacteria in the step S1 for 24-48h at 35-40 ℃, adding the culture solution into an alcohol sample, and screening the alcohol effect dispelling capacity; s3, centrifuging the alcohol sample in the step S2, taking the upper solution, and repeating the steps according to S1 and S2 again until proper spore bacteria are bred. According to the method, the spore bacteria are cultivated firstly, and then the antialcoholism capability is screened, so that the strains with high antialcoholism capability are gradually bred, and the operation is efficient and rapid; the breeding device can perform decomposition contrast screening of different alcoholic strength under the same environment, and breeding efficiency is improved.)

1. A selective breeding method of spore bacteria for alcohol decomposition is characterized by comprising the following steps:

s1, inoculating the liquid strains of the spore bacteria to a culture medium in a breeding device for cultivation, wherein the inoculation amount is 6-10%; the culture medium comprises the following raw materials in parts by weight: 30-50g/L corn flour, 5-8g/L glucose, 5-10g/L peptone, 20-40g/L dragon claw fruit powder, 20-30g/L kudzu root powder, 0.1-0.2g/L magnesium sulfate, 0.3-0.8g/L sodium dihydrogen phosphate and 0.1-0.3g/L sodium sulfite;

s2, culturing for 24-48h at 35-40 ℃ in the step S1, then adding a proper amount of culture solution into an alcohol sample, and screening the alcohol effect dispelling capacity;

s3, centrifuging the alcohol sample in the step S2, taking the upper solution, repeating the steps S1 and S2 again for breeding until proper spore bacteria are bred.

2. The method for selectively breeding the spore bacteria for alcohol decomposition as claimed in claim 1, wherein the pH of the culture medium of step S1 is 7-7.3, the spore bacteria is bacillus, and the viable bacteria content of the spore bacteria used during inoculation is more than 30-60 hundred million/mL.

3. The method for selectively breeding spore bacteria for alcohol decomposition as claimed in claim 2, wherein the viable bacteria content of the spore bacteria culture solution of step S2 exceeds 500-700 hundred million/mL.

4. The method for breeding the spore bacteria for decomposing the alcohol according to any one of the claims 1 to 3, characterized in that the breeding device comprises a tank body (1), and a cover body (11) is arranged on the tank body (1); the utility model discloses a culture tube assembly, including jar body (1), be provided with culture tube assembly (2) on the jar body (1), culture tube assembly (2) are including first culture tube (21), one side of first culture tube (21) is provided with second culture tube (22).

5. The method for selective breeding of spore bacteria for alcohol decomposition according to claim 4, wherein the first culture tube (21) and the second culture tube (22) are connected through a pipeline (23), and a one-way electromagnetic valve (24) is arranged on the pipeline (23) so that the culture solution in the first culture tube (21) can flow into the second culture tube (22).

6. The method for breeding the spore bacteria for decomposing the alcohol according to the claim 5, characterized in that a shell (3) is arranged outside the tank body (1), a rotating shaft (31) is arranged in the shell (3), and the rotating shaft (31) is connected with the tank body (1) in a buckling way; the rotating shaft (31) is connected with the rotating device (32) arranged on the outer side of the shell (3); and a drain pipe (33) is arranged on the outer side of the shell (3).

7. A method for selectively breeding spore bacteria for alcohol decomposition as claimed in claim 6, characterized in that the shell (3) is provided with a shell cover (34) for covering the tank (3) inside the shell.

8. A method for selectively breeding spore bacteria for alcohol decomposition as claimed in claim 7, characterized in that a support plate (35) is arranged in the housing (3), the top of the support plate (35) is connected with the tank (1) to make the tank (1) and the housing (3) not in complete contact.

9. The method for breeding spore bacteria for alcohol decomposition according to claim 8, wherein the cultivating tube assembly (2) is provided with a plurality of groups, the first cultivating tube (21) is used for loading a culture medium to cultivate the spore bacteria, and the second cultivating tube (22) is used for loading an alcohol sample to test the capacity of the spore bacteria to decompose alcohol.

10. The method for selectively breeding spore bacteria for alcohol decomposition as claimed in claim 9, wherein the alcohol degree of the alcohol sample loaded in the second culture tube (22) of each set of the culture tube assembly (2) is gradually increased, and the alcohol sample is used according to the sequence from low concentration to high concentration.

Technical Field

The invention belongs to the technical field of microbial breeding, and particularly relates to a method for breeding spore bacteria for alcohol decomposition.

Background

The main component of the wine is alcohol, and excessive drinking and long-term drinking not only cause harm to the health of human bodies, but also threaten the safety of public society by behaviors such as drunk driving and the like. The existing anti-inebriation product is mainly made of Chinese herbal medicines, the price is relatively high, and the components of the Chinese herbal medicines are relatively complex and are difficult to be widely applied. Therefore, it is very important to develop a rescue product with wide application population.

The bacillus is a bacillus or coccus capable of forming spores (endospore), and includes bacillus, lactobacillus, clostridium, desthioenterococcus, and sporosarcina, etc. They have strong resistance to external harmful factors and wide distribution, and exist in soil, water, air, animal intestinal tracts and the like. Researches show that the spore bacteria can decompose alcohol, so that the strain with strong capability of relieving alcoholism has a positive effect on the development of a product for relieving alcoholism by breeding.

Disclosure of Invention

The invention provides a method for breeding spore bacteria for alcohol decomposition, which aims to solve the technical problems.

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

a method for breeding spore bacteria for alcohol decomposition, which comprises the following steps:

s1, inoculating the liquid strains of the spore bacteria to a culture medium in a breeding device for cultivation, wherein the inoculation amount is 6-10%; the culture medium comprises the following raw materials in parts by weight: 30-50g/L corn flour, 5-8g/L glucose, 5-10g/L peptone, 20-40g/L dragon claw fruit powder, 20-30g/L kudzu root powder, 0.1-0.2g/L magnesium sulfate, 0.3-0.8g/L sodium dihydrogen phosphate and 0.1-0.3g/L sodium sulfite;

s2, culturing for 24-48h at 35-40 ℃ in the step S1, then adding a proper amount of culture solution into an alcohol sample, and screening the alcohol effect dispelling capacity;

and S3, centrifuging the alcohol sample in the step S2, taking the upper solution, and repeating the steps S1 and S2 again until proper spore bacteria are bred.

Further, the pH of the culture medium in the step S1 is 7-7.3, the spore bacteria are bacillus, and the content of viable bacteria of the spore bacteria used during inoculation is more than 30-60 hundred million/mL.

Further, the viable bacteria content of the spore bacteria culture solution of the step S2 exceeds 500-700 hundred million/mL.

Further, the breeding device comprises a tank body, and a cover body is arranged on the tank body; the jar is provided with the cultivation pipe subassembly on the body, the cultivation pipe subassembly includes first cultivation pipe, one side of first cultivation pipe is provided with the second cultivation pipe. The first culture tube is provided with a culture medium for culturing thalli, and the second culture tube is used for loading an alcohol sample for carrying out alcohol disintoxication capability screening on the cultured thalli.

Furthermore, the first culture tube and the second culture tube are connected through a pipeline, and a one-way electromagnetic valve is arranged on the pipeline so that the culture solution in the first culture tube can flow into the second culture tube. The arranged pipeline is beneficial to transferring the cultured culture solution containing the spore bacteria into an alcohol sample for carrying out the screening of the capability of dispelling the effects of alcohol, and the operation is quick; the one-way valve is arranged to ensure that the solution can only flow from the first culture tube into the second culture tube.

Further, a shell is arranged on the outer side of the tank body, a rotating shaft is arranged in the shell and is connected with the tank body in a buckling mode; the rotating shaft is connected with the rotating device arranged on the outer side of the shell; and a drain pipe is arranged on the outer side of the shell. The rotating device and the rotating shaft can drive the tank body to rotate during operation, thereby being beneficial to the full contact of the culture medium and the strains and improving the propagation efficiency. Water can be added into the shell to heat the tank body in a water bath manner, so that the temperature in the tank body accords with the growth and the propagation of strains.

Further, a housing cover 34 is disposed on the housing 3 for covering the tank 3 inside the housing. A supporting plate 35 is arranged in the shell (3), and the top of the supporting plate 35 is connected with the tank body 1, so that the tank body 1 is not completely contacted with the shell 3.

Further, the culture tube assembly is provided with a plurality of groups, the first culture tube is used for loading a culture medium to culture the spore bacteria, and the second culture tube is used for loading an alcohol sample to test the capacity of the spore bacteria to decompose alcohol. First culture tube loads the culture medium and is used for the spore fungus, and the second culture tube loads the alcohol sample, when the fungus live bacteria volume of the interior cultivation of first culture tube reaches certain concentration, carries out the alcohol decomposition screening in adding alcohol through the culture solution, can improve operating efficiency, and convenient operation is swift simultaneously.

Further, the alcohol degree of the alcohol sample loaded in the second culture tube of each group of the culture tube assemblies is gradually increased, and the alcohol sample is used according to the sequence from low concentration to high concentration. During breeding, the alcohol sample is set to be of gradually increased concentration, so that the spore bacteria can gradually adapt to the alcohol sample, the survival rate of the spore bacteria is ensured, and the strains with high alcohol decomposition performance are bred.

The invention has the advantages that: 1. the bacillus is cultivated firstly, and then the antialcoholism capability is screened, so that strains with high antialcoholism capability are gradually bred, and the operation is efficient and rapid; 2. the breeding device used by the invention can perform decomposition contrast screening of different alcoholic strength under the same environment, improves breeding efficiency, automatically guides the bacterial liquid into the alcohol sample through a pipeline in the breeding process, avoids external interference caused by manual treatment, and has high efficiency and rapidness in operation.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a schematic structural diagram of the present invention.

FIG. 2 is a top view of the can body.

FIG. 3 is a schematic view of the structure of the culture tube assembly.

Reference numerals: 1-tank, 11-cover, 12-sampling port, 2-culture tube component, 21-first culture tube, 22-second culture tube, 23-pipeline, 24-one-way solenoid valve, 3-shell, 31-rotating shaft, 32-rotating device, 33-drainage tube, 34-shell cover and 35-supporting plate.

Detailed Description

In order to facilitate a better understanding of the invention, reference is made to the following examples, which are set forth to illustrate, but are not to be construed as the limit of the invention.

Example 1

A method for breeding spore bacteria for alcohol decomposition comprises the following steps:

s1, inoculating the liquid strains of the spore bacteria to a culture medium in a breeding device for cultivation, wherein the inoculation amount is 6%; the culture medium comprises the following raw materials in parts by weight: 30g/L of corn flour, 5g/L of glucose, 5g/L of peptone, 20g/L of dragon claw fruit powder, 20g/L of radix puerariae powder, 0.1g/L of magnesium sulfate, 0.3g/L of sodium dihydrogen phosphate and 0.1g/L of sodium sulfite;

s2, culturing for 24 hours in the step S1 at the temperature of 35 ℃, then adding a proper amount of culture solution into an alcohol sample, and screening the alcohol effect dispelling capacity;

and S3, centrifuging the alcohol sample in the step S2, taking the upper solution, and repeating the steps S1 and S2 again until proper spore bacteria are bred.

And (3) the pH value of the culture medium of the step S1 is 7, the spore bacteria are bacillus, and the content of live bacteria of the spore bacteria used during inoculation is more than 30 hundred million/mL.

The viable bacteria content of the spore bacteria culture solution of the step S2 exceeds 500 hundred million/mL.

As shown in fig. 1-3, the breeding device comprises a tank body 1, a cover body 11 is arranged on the tank body 1; a culture tube assembly 2 is arranged on the tank body 1, the culture tube assembly 2 comprises a first culture tube 21, and a second culture tube 22 is arranged on one side of the first culture tube 21; be provided with sample connection 12 on the lid 11, sample connection 12 is provided with a plurality of to for, first culture tube 21, second culture tube 22 for, the sample connection that sets up is used for the sample to detect the condition that living fungus quantity and bacterial divide-joint to alcohol. The sampling port 12 is provided with a cover, and is covered with the cover after use.

As shown in FIGS. 1 to 3, the first culture tube 21 and the second culture tube 22 are connected to each other through a pipe 23, and a one-way solenoid valve 24 is provided on the pipe 23 to allow the culture solution in the first culture tube 21 to flow into the second culture tube 22.

As shown in fig. 1, a shell 3 is arranged outside the can body 1, a rotating shaft 31 is arranged in the shell 3, and the rotating shaft 31 is connected with the can body 1 in a snap fit manner; the rotating shaft 31 is connected with a rotating device 32 arranged outside the shell 3; a drain pipe 33 is provided outside the casing 3.

As shown in fig. 1, the housing 3 is provided with a housing cover 34 for covering the can 3 inside the housing. A supporting plate 35 is arranged in the shell (3), and the top of the supporting plate 35 is connected with the tank body 1, so that the tank body 1 is not completely contacted with the shell 3.

The culture tube assembly 2 is provided with a plurality of groups, a first culture tube 21 is used for loading culture medium to culture the spore bacteria, and a second culture tube 22 is used for loading an alcohol sample to test the capacity of the spore bacteria to decompose alcohol.

The alcohol concentration of the alcohol sample loaded in the second culture tube 22 of each group of the culture tube assemblies 2 is gradually increased, and the alcohol sample is used according to the sequence from low concentration to high concentration.

The use method of the breeding device comprises the following steps: before use, the culture medium, the tank body and the like are sterilized to avoid the influence of mixed bacteria on breeding; opening the shell cover, opening the cover body, adding the culture medium into the first culture tube, inoculating the liquid strains of the spore bacteria onto the culture medium, adding the alcohol sample into the second culture tube, covering the cover body, covering the cover shell, and adding proper hot water into the shell body according to the requirement to carry out water bath; in the cultivation process, the rotating device is started to enable the tank body to swing, so that thalli are fully contacted with a culture medium, and the reproduction efficiency of the thalli is improved; when the viable bacteria amount in the first culture tube meets the requirement, opening the one-way electromagnetic valve to enable the culture solution in the first culture tube to enter the second culture tube, and screening the antialcoholism capability; during the cultivation process, the strain quantity in the culture solution and the antialcoholism capability of the strain can be detected by sampling at the sampling port.

Example 2

A method for breeding spore bacteria for alcohol decomposition comprises the following steps:

s1, inoculating the liquid strains of the spore bacteria to a culture medium in a breeding device for cultivation, wherein the inoculation amount is 8%; the culture medium comprises the following raw materials in parts by weight: 40g/L corn flour, 6g/L glucose, 7g/L peptone, 30g/L dragon's claw fruit powder, 25g/L kudzu vine root powder, 0.1g/L magnesium sulfate, 0.5g/L sodium dihydrogen phosphate and 0.2g/L sodium sulfite;

s2, culturing for 35h in the step S1 at 38 ℃, then adding a proper amount of culture solution into an alcohol sample, and screening the alcohol disintoxicating capability.

And S3, centrifuging the alcohol sample in the step S2, taking the upper solution, and repeating the steps S1 and S2 again until proper spore bacteria are bred.

The pH value of the culture medium of the step S1 is 7.2, the spore bacteria are bacillus, and the content of viable bacillus used in inoculation is more than 50 hundred million/mL.

The viable bacteria content of the spore bacteria culture solution of the step S2 exceeds 600 hundred million/mL.

Example 3

A method for breeding spore bacteria for alcohol decomposition comprises the following steps:

s1, inoculating the liquid strains of the spore bacteria to a culture medium in a breeding device for cultivation, wherein the inoculation amount is 10%; the culture medium comprises the following raw materials in parts by weight: 50g/L corn flour, 8g/L glucose, 10g/L peptone, 40g/L dragon claw fruit powder, 30g/L kudzu vine root powder, 0.2g/L magnesium sulfate, 0.8g/L sodium dihydrogen phosphate and 0.3g/L sodium sulfite;

s2, culturing for 48h at 40 ℃ in the step S1, then adding a proper amount of culture solution into an alcohol sample, and screening the alcohol disintoxicating capability.

And S3, centrifuging the alcohol sample in the step S2, taking the upper solution, and repeating the steps S1 and S2 again until proper spore bacteria are bred.

The pH value of the culture medium of the step S1 is 7.3, the spore bacteria are bacillus, and the content of viable bacillus used in inoculation is over 60 hundred million/mL.

The viable bacteria content of the spore bacteria culture solution of the step S2 exceeds 700 hundred million/mL.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. The term "comprising" is used to specify the presence of stated elements, but not to preclude the presence or addition of additional like elements in a process, method, article, or apparatus that comprises the stated elements.

The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.