Non-contact continuous multipoint sampling laboratory small fermentation device

Abstract

The utility model discloses a non-contact continuous multipoint sampling laboratory small fermentation device, and belongs to the technical field of small microbiological fermentation devices. The device consists of a conical flask with a silica gel plug and a one-way ventilation valve with an inner wall pipe, the inner wall pipe is arranged in the one-way ventilation valve; an insertion hole is formed in the middle of the silica gel plug, and the inner wall tube extends into the conical flask through the insertion hole; an inner wall cover is arranged on the outer side of an inlet, located in the one-way ventilation valve, of the inner wall pipe, and an outer wall cover is arranged at the top end of the one-way ventilation valve. A material taking pipe is arranged between the one-way ventilation valve and the conical flask, an air end inlet of the material taking pipe is located outside the top end of the one-way ventilation valve, and the material taking pipe naturally droops in the one-way ventilation valve, then penetrates through a gap between the inner wall cover and the inner wall pipe from bottom to top, then penetrates through the inner wall pipe from top to bottom and finally extends below the liquid level of fermentation liquid in the conical flask. According to the utility model, the contradiction between uncovering material taking and continuous fermentation in the small fermentation process is effectively solved, and non-contact continuous material taking is realized.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the technical field of small microbiology fermentation device, specifically relates to a kind of laboratory small formula fermentation device of non-contact continuous multi-point sampling. BACKGROUND

[0002] Anaerobic fermentation is applied in many fields, such as yeast brewing beer, but the industrial brewing equipment of brewery etc. is generally large fermentation tank, and laboratory does not need to use such large-capacity fermentation tank, so convenient small fermentation device is a good choice. Small fermentation has wide application in life, production and experimental research, such as production of alcoholic beverage, bread, steamed bun and other food microbiological physiological research, simulation of microbial growth and metabolism process, research on physiological characteristics, growth law and metabolism mechanism of microorganism. In addition, small fermentation can also be used for producing biofuels, such as biodiesel, ethanol, etc., to provide technical support for the development and utilization of renewable energy.

[0003] Beer is the third largest consumer beverage after water and tea, and occupies an important position in our daily drinks. With the increase of people's economic income and the youth of consumer population, the demand of consumers for beer also starts to change to high quality, individualization and diversification, and improving the intrinsic quality of beer, especially improving the flavor of beer and enhancing the nutritional value of beer, is an important research direction to improve the quality and profit of beer in China. In the beer fermentation process, brewing yeast anaerobic fermentation converts sugar into alcohol and carbon dioxide gas, and factors such as wort quality, inorganic ions, osmotic pressure, temperature, pH will have a certain influence on the physiological activity of brewing yeast. In order to explore the growth of yeast in the fermentation process and the changes of alcohol content, pH, various sugar concentrations and other wine quality, sampling at multiple specific time points is needed, and sampling usually needs to open the cover, but each time opening the cover will introduce external oxygen, affect the gas pressure in the container, and then affect the anaerobic fermentation process; Generally, the laboratory small fermentation can only meet the fermentation process, and cannot sample without destroying the anaerobic conditions in the container, that is, there is a contradiction between fermentation and continuous sampling, and the utility model can effectively solve this contradiction. UTILITY MODEL CONTENT

[0004] The utility model aims at providing a kind of laboratory small formula fermentation device of non-contact continuous multi-point sampling, which can solve the contradiction between opening cover sampling and continuous fermentation, can monitor fermentation condition and carry out sampling, simple and convenient operation, and save resources.

[0005] The utility model discloses a kind of non-contact continuous multi-point sampling laboratory small type fermentation device of taking, it is made of flask with silica gel plug and one-way air valve with inner wall pipe;Silica gel plug is tightly embedded in the bottle mouth of flask, avoid the gap between silica gel plug and the bottle mouth of flask, then use sealing film to fix silica gel plug and the bottle mouth of flask, can further prevent other substances in air from polluting flask;Inner wall pipe is arranged in the inside of one-way air valve, and it is airtight with the bottom outer wall of one-way air valve;There is straight direction jack in the middle of silica gel plug, inner wall pipe is inserted into flask through the jack, so that one-way air valve is located directly above flask;The outlet of inner wall pipe in flask is located above the liquid level of fermentation broth in flask, inner wall cover is arranged on the outside of the inlet of inner wall pipe in one-way air valve, and there is a certain gap between inner wall cover and inner wall pipe inlet;Outer wall cover is arranged at the top of one-way air valve, outer wall cover is covered when fermentation, and outer wall cover is removed when sampling;Sampling pipe (soft silica gel material) is arranged between one-way air valve and flask, air end inlet of sampling pipe is located at the top outside of one-way air valve, small clip is arranged on the air end inlet of sampling pipe, sampling pipe naturally droops in one-way air valve, then pass through the gap between inner wall cover and inner wall pipe from bottom to top, pass through inner wall pipe from top to bottom, and finally extend below the liquid level of fermentation broth in flask.

[0006] The specific engineering process of the utility model is as follows:

[0007] 1, one-way air valve, inner wall pipe, inner wall cover, outer wall cover, silica gel plug and flask (one-way air valve, inner wall pipe, inner wall cover, outer wall cover are plastic material, silica gel plug is silica gel material, flask is glass material) are subjected to 121 DEG C, 20min high-pressure sterilization;

[0008] 2, pour wort or other liquid fermentation raw materials into flask (flask specification is 150mL, can be adjusted according to actual needs, 150mL specification flask can pour 90-105mL wort to ferment, that is, fermentation broth accounts for 60-70% of flask volume) in super-clean workbench, realize the "canning" process of fermentation;

[0009] 3. Insert the outlet of the inner tube into the hole of the silicone plug. Cover the inlet of the inner tube with the inner cap (the inner diameter of the inner cap should be larger than the outer diameter of the inlet of the inner tube). Inject sterilized purified water into the one-way vent valve to achieve "liquid seal". If no material is needed, simply cover with the outer cap and allow fermentation for a certain period of time. If continuous material is needed, the outer cap is not required. Cut an appropriate length of material collection tube, and before injecting purified water, extend the tube downwards along the inner tube. Stop inserting the tube 2-3 cm below the surface of the fermentation broth in the conical flask, cover it with the inner cap, and adjust the curvature of the sampling tube so that it is not squeezed by the inner cap during sampling. Then inject sterilized purified water into the one-way vent valve to achieve "liquid seal". When sampling, use a clean disposable syringe to draw the fermentation broth as needed at the entrance of the sampling tube. After drawing, rinse the entrance of the sampling tube with 75% ethanol. Finally, clamp the air end entrance of the sampling tube with a small clamp before fermentation.

[0010] Furthermore, due to evaporation, the pure water used for liquid sealing may evaporate, so it is necessary to pay close attention to the water level changes of the one-way vent valve and add water from time to time to maintain the pressure difference between the inside and outside.

[0011] The beneficial effects of this utility model are: Figure 2 As shown, CO2 is produced during the anaerobic fermentation of the fermentation broth in the conical flask. 2 After passing through the inner wall tube, inner wall cover, and purified water, the gas can be effectively discharged from the top of the one-way vent valve (there is a gap between the outer wall cover and the one-way vent valve, allowing gas to be discharged in one direction), thus achieving one-way ventilation of the device. Furthermore, it effectively resolves the contradiction between opening the lid for material collection and continuous fermentation during small-scale fermentation, achieving contactless continuous material collection. Attached Figure Description

[0012] Figure 1 This is a schematic diagram of the small-scale fermentation device described in this utility model during fermentation.

[0013] Figure 2 This is a schematic diagram of the small-scale fermentation device of this utility model when fermenting and collecting materials simultaneously;

[0014] The names of each part are: 1. One-way vent valve, 2. Inner wall tube, 3. Silicone stopper, 4. Conical flask, 5. Inner wall cap, 6. Outer wall cap, 7. Small clip, 8. Sampling tube.

[0015] Table 1 shows the OD values ​​of the fermentation broths from Examples 1 and 2. 600 Value data;

[0016] Detailed Implementation

[0017] The utility model will be further described below in combination with examples. The purpose of combining examples is to elaborate the utility model in detail, not to limit the utility model.

[0018] Example 1: Anaerobic culture of strain expansion scale

[0019] As shown in Figure 1 , the to-be-fermented strain of Pasteur yeast CBS1513 (which can be directly purchased) activated in advance to the logarithmic phase is prepared, the one-way air valve 1, the inner wall pipe 2, the inner wall cover 5, the outer wall cover 6 and the silica gel plug 3 and the conical flask 4 are subjected to high-pressure sterilization at 121 DEG C for 20 min; 12°P wort (Guo Xian Guo. Brewing Wine Technology [M]. Brewing Wine Technology, 1996) is poured into the conical flask 4 in the super-clean workbench, and the pouring amount of the wort is 30% of the volume of the conical flask 4, thus realizing the "canning" process of fermentation; the sterilized inner wall pipe 2 is inserted into the insertion hole in the middle part of the silica gel plug 3, the silica gel plug 3 is tightly embedded into the bottle mouth of the conical flask 4, so as to avoid the air gap between the silica gel plug 3 and the bottle mouth of the conical flask 4, and the sealing film is used to fix the bottle mouth of the conical flask 4 and the silica gel plug 3; then the inner wall cover 5 is covered, sterilized pure water is injected into the one-way air valve 1 to realize "liquid sealing", and finally, 28 DEG C fermentation is carried out (the conical flask 4 is placed on the shaking bed with a pre-set temperature, the shaking bed speed is set to 150 r / min, and the setting speed can mix the fermentation liquid, which is equivalent to stirring).

[0020] Example 2: Small-scale fermentation of common Saccharomyces cerevisiae

[0021] As shown in Figure 2As shown, the fermenting strain of Pasteurella CBS1513 (can be purchased directly) in the logarithmic phase is prepared to be activated in advance, a one-way air valve 1, an inner wall tube 2, an inner wall cover 5 and a silica gel plug 3, a conical flask 4, a sampling tube 8 are subjected to high-pressure sterilization at 121 ℃ for 20 min, 12°P wort (Guo Xian Guo. Brewing Wine Technology [M]. Brewing Wine Technology, 1996) is poured into the conical flask 4 in a super-clean bench, the pouring amount of the wort is 30% of the volume of the conical flask 4, and the "canning" process of fermentation is realized; the sterilized inner wall tube 2 is inserted into the insertion hole in the middle part of the silica gel plug 3, the silica gel plug 3 is tightly embedded into the bottle mouth of the conical flask 4, and the silica gel plug 3 and the conical flask 4 are fixed by a sealing film; a soft silica gel tube with an appropriate length is cut as the sampling tube 8, the sampling tube 8 is naturally hung in the one-way air valve 1, then passes through the gap between the inner wall cover 5 and the inner wall tube 2 from bottom to top, then passes through the inner wall tube 2 from top to bottom, and finally extends into the fermentation liquid surface of the conical flask 4 by 2 cm and stops, the inner wall cover 5 is covered, the bending degree of the sampling tube 8 is adjusted so that the sampling tube 8 is not pressed by the inner wall cover 5 during sampling, finally the inner wall cover 5 is covered, and sterilized pure water is injected into the one-way air valve 1 to realize "liquid sealing"; 28 ℃ fermentation is carried out (the conical flask 4 is placed on a pre-set temperature shaker, the rotation speed of the shaker is set to 150 r / min, and the rotation speed is set to mix the fermentation liquid, which is equivalent to stirring).

[0022] Due to the evaporation effect, the pure water used for liquid sealing may volatilize, so it is necessary to pay attention to the water level change in time and add water at irregular times to maintain the difference between the inner and outer air pressures.

[0023] As shown in Table 1, the OD values of the fermentation liquids of the fermentation experiment of Example 1 (corresponding to curve A in the figure) and the fermentation experiment while sampling of Example 2 (corresponding to curve B in the figure) are measured (a hand-held device for directly measuring OD values is used to directly measure the conical flask, and the fermentation liquid body can also be measured by using a spectrophotometer, an enzyme marker and the like), the abscissa is time, and the ordinate is the OD value; there is no obvious difference between the two curves, and it can be seen that the device can realize non-contact multiple sampling and ensure that the bacteria continuously ferment under anaerobic conditions. 600 600 600

[0024] ​​​The above is only a preferred embodiment of the present application, and does not limit the scheme of the present application in any form. Any simple modification, equivalent change and modification of the above embodiment according to the method of the present application fall within the protection scope of the present application.

Claims

1. A non-contact, continuous multi-point sampling laboratory benchtop fermentation device, characterized by: The application is composed of a conical bottle (4) with a silica gel plug (3) and a one-way air valve (1) with an inner wall tube (2); the silica gel plug (3) is tightly embedded in the bottle mouth of the conical bottle (4); the inner wall tube (2) is arranged inside the one-way air valve (1) and is tightly installed with the outer wall of the bottom of the one-way air valve (1); a straight direction jack is arranged in the middle of the silica gel plug (3) and the inner wall tube (2) is inserted into the conical bottle (4) through the jack; an inner wall cover (5) is arranged outside the entrance of the inner wall tube (2) in the one-way air valve (1) and there is a certain gap between the inner wall cover (5) and the entrance of the inner wall tube (2); an outer wall cover (6) is arranged at the top end of the one-way air valve (1) and the outer wall cover (6) is covered when fermentation is carried out and the outer wall cover (6) is removed when material is taken; a material taking tube (8) is arranged between the one-way air valve (1) and the conical bottle (4) and the air end entrance of the material taking tube (8) is located outside the top end of the one-way air valve (1); a small clip (7) is arranged on the air end entrance of the material taking tube (8); the material taking tube (8) naturally drops in the one-way air valve (1), then passes through the gap between the inner wall cover (5) and the inner wall tube (2) from bottom to top, then passes through the inner wall tube (2) from top to bottom and finally is inserted into the fermentation liquid below the liquid surface in the conical bottle (4).

2. A non-contact, continuous multi-point sampling laboratory fermenter as claimed in claim 1, wherein: The silica gel plug (3) and the bottle mouth of the conical bottle (4) are fixed by a sealing film.

3. A non-contact, continuous multi-point sampling laboratory fermenter as claimed in claim 1, wherein: The fermentation liquid accounts for 60-70% of the volume of the conical bottle (4).

4. A non-contact, continuous multi-point sampling laboratory fermenter as claimed in claim 1, wherein: The outlet of the material taking tube (8) is inserted into the fermentation liquid below the liquid surface in the conical bottle (4) by 2-3 cm along the inner wall tube (2).

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