CN212341119U - Nitrogen fixation enzyme activity measuring device improved by acetylene reduction method - Google Patents
Nitrogen fixation enzyme activity measuring device improved by acetylene reduction method Download PDFInfo
- Publication number
- CN212341119U CN212341119U CN202021048209.2U CN202021048209U CN212341119U CN 212341119 U CN212341119 U CN 212341119U CN 202021048209 U CN202021048209 U CN 202021048209U CN 212341119 U CN212341119 U CN 212341119U
- Authority
- CN
- China
- Prior art keywords
- valve
- nutrient solution
- rubber plug
- inlet pipe
- sealed reaction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The utility model discloses a nitrogenase activity measuring device improved by an acetylene reduction method, which comprises a sealed reaction bottle, wherein the mouth of the sealed reaction bottle is provided with an isobutyl rubber plug, a gas sample inlet pipe and a nutrient solution sample inlet pipe are fixed on the isobutyl rubber plug, the gas sample inlet pipe is sequentially connected with a first valve and a second valve, and the other end of the gas sample inlet pipe is communicated with an injector through a rubber plug; the nutrient solution sampling pipe is sequentially provided with a third valve and a multi-connection pipe valve; the multi-connection pipe valve is connected with a nutrient solution bottle; the sealed reaction bottle is positioned in the porous heat-preservation type vibration plastic component. The utility model solves the problem of air leakage caused by pinholes in the process of measuring the activity of azotase, reduces the system error of each reaction system and improves the reproducibility of experimental results; meanwhile, due to the design of the multi-joint valve, an operator can finish more sample pretreatment in batches within a certain time, and the experimental efficiency is improved.
Description
Technical Field
The utility model relates to a testing arrangement improvement field specifically is an utilize nitrogenase activity survey device of acetylene reduction method improvement for accurate high-efficient assay nitrogenase activity.
Background
The acetylene reduction method is one of the commonly used detection methods, and the method is to indirectly determine whether the azotobacter activity exists and the level of the azotobacter activity by detecting whether acetylene is reduced to form ethylene and the amount of the ethylene through a gas chromatograph according to the principle that the azotobacter has substrate diversity. At present, most of the activities of nitrogen-fixing enzymes of soil or plant nodules are measured by an acetylene reduction method in laboratories. At present, devices for measuring the activity of azotobacter by acetylene reduction methods are still not mature.
Most of the existing experimental devices are simple, and have many defects, such as: the problem of air leakage of the rubber plug caused by a pinhole in the process of measuring the activity of the azotobacter cannot be well avoided; the pretreatment of the samples can not be carried out simultaneously, and the pretreatment of a single sample takes a long time; the culture process is affected by the shape of soil and temperature difference, and acetylene reduction reaction is insufficient.
SUMMERY OF THE UTILITY MODEL
The utility model aims at: the optimized device for determining the activity of the azotobacter by using the acetylene reduction method is used for accurately and efficiently measuring the activity of the azotobacter in soil, root nodules and the like.
The technical scheme of the utility model is that:
the azotase activity measuring device improved by the acetylene reduction method comprises a sealed reaction bottle, wherein an isobutyl rubber plug is arranged at the mouth of the sealed reaction bottle, a gas sampling tube and a nutrient solution sampling tube are fixed on the isobutyl rubber plug, the gas sampling tube is sequentially connected with a first valve and a second valve, and the other end of the gas sampling tube is communicated with an injector through a rubber plug;
the nutrient solution sampling pipe is sequentially provided with a third valve and a multi-connection pipe valve; the multi-connection pipe valve is connected with a nutrient solution bottle;
the sealed reaction bottle is positioned in the porous heat-preservation type vibration plastic component.
The inside of the sealed reaction bottle is filled with a sample needing to be measured for the activity of the azotobacter; the porous heat-preservation type vibration plastic component is used for placing the sealed reaction bottle in a vibration incubator; the porous heat-preservation type vibration plastic component can be used for placing a plurality of reaction bottles at the same time, and is convenient to take; meanwhile, the temperature can be kept to reduce the temperature difference in each reaction bottle in the experiment; the same-frequency oscillation can promote the reaction to be more sufficient and thorough, and reduce the system error.
The multi-connection pipe valve can connect a plurality of sample nutrient solution inlet pipes with the nutrient solution bottle and simultaneously input nutrient solution.
The device for measuring the activity of the nitrogenase improved by using the acetylene reduction method comprises the following steps:
A) taking 1g of fresh soil or plant root nodule 10g, and filling into a 100ml sealed reaction bottle;
B) covering an isobutyl rubber plug with a nutrient solution sampling pipe and a gas sampling pipe;
C) 2ml of 0.1mol/l glucose nutrient solution is added into the sealed reaction bottle through a nutrient solution sampling tube, and a third valve is closed;
D) connecting the syringe with a rubber plug, opening the second valve, keeping the first valve closed, and sucking air in the gas sample introduction pipe until the pipe is shriveled;
E) closing the second valve, taking down the syringe to discharge air in the tube, and connecting the rubber plug again;
F) simultaneously opening the first valve and the second valve, sucking 10ml of air, closing the first valve and the second valve, and taking down the syringe to discharge the air in the tube;
G) pumping 10ml of acetylene gas with the purity of 99.9 percent by using an injector, connecting a rubber plug, sequentially opening a second valve and a first valve of a valve, injecting the gas into a sealed reaction bottle, and then rapidly closing the second valve and the first valve of the valve;
H) placing the sealed reaction bottle in a porous heat-preservation type vibration plastic component, and culturing for 24 hours in a constant-temperature vibration incubator at 28 ℃;
I) opening the second valve, using the injector to suck to discharge the air in the gas sampling pipe, connecting the rubber plug again, and opening the first valve to suck 10ml of reaction gas for determination;
J) ethylene (C) was determined by gas chromatography (GC, Agilent7890B, USA)2H4) The gas chromatograph adopts a capillary column (DB-WAX, 122-7032, Agilent, USA) and an FID detector
Before injecting acetylene gas into reaction system, connect the rubber buffer with the syringe, open the second valve and close first valve, the intraductal air of suction gas sampling becomes shrivelled in order to reduce reaction system foreign gas to the pipe, improves experimental result accuracy and degree of accuracy.
Can combine the timing function of the incubator to obtain the measured ethylene (C)2H4) Yield to obtain nitrogenase activity (rate of ethylene reduction per unit time of soil/nodule) data.
The syringe 1 for sucking gas should be exclusively used for gas.
Compared with the prior art, the utility model discloses following beneficial effect has:
(1) the utility model discloses can carry out a plurality of sample pretreatments simultaneously, avoid taking traditional syringe to prick the loaded down with trivial details operation of hole with the nutrient solution.
(2) The utility model discloses can directly realize closing of pipeline through the valve of PVC hose, realize communicating with each other of serum bottle and syringe, reduce and prick the rubber buffer with the needle tubing of taking the syringe needle and lead to its device gas leakage that causes that damages to the influence of reaction system, improve experimental data's accuracy and reproducibility.
(3) The system of the utility model has the advantages of simple structure, accurate measuring result, high testing speed and low cost, and provides favorable help for analyzing and researching azotase, microorganism gas production degree, azotization efficiency and the like.
Drawings
Fig. 1 is a schematic perspective view of the present invention;
FIG. 2 is a schematic structural view of the present invention;
fig. 3 is a schematic view of the porous heat-insulating vibrating plastic component of the present invention.
Detailed Description
The test materials used in the following examples were all commercially available unless otherwise specified. As shown in fig. 1 and 2, the device for measuring the activity of a nitrogenase improved by using an acetylene reduction method comprises a sealed reaction bottle 7, wherein an isobutyl rubber plug 6 is arranged at the mouth of the sealed reaction bottle 7, a gas sampling tube 4 and a nutrient solution sampling tube 9 are fixed on the isobutyl rubber plug 6, the gas sampling tube 4 is sequentially connected with a first valve 5 and a second valve 3, and the other end of the gas sampling tube 4 is communicated with an injector 1 through a rubber plug 2;
the nutrient solution sampling pipe 9 is sequentially provided with a third valve 10 and a multi-connection pipe valve 11; the multi-joint valve 11 is connected with a nutrient solution bottle 12;
the sealed reaction bottle 7 is positioned in the porous heat-preservation type vibration plastic component 8.
A sample needing to be measured for the activity of the azotobacter is contained in the sealed reaction bottle 7; the porous heat preservation type oscillation plastic component 8 is used for placing a sealed reaction bottle in an oscillation incubator, as shown in figure 3; the multi-joint valve 11 can connect a plurality of sample nutrient solution feeding pipes together and simultaneously feed nutrient solution. The nutrient solution sampling pipe 9 is a PVC hose with the diameter of only 1mm, so that the residue of gas or liquid on the pipe wall can be effectively reduced, and the experimental accuracy is improved.
The gas sample inlet pipe 4 of the injector 1 connected with the sealed reaction bottle 7 is a PVC hose with the diameter of 1 mm; the device comprises two gas stop valves, wherein the second valve 3 close to the injector 1 is opened, and the first valve 5 close to the sealed reaction bottle 7 is closed, so that impurity gas in the device system can be effectively discharged; the gas replacement can be accurately carried out by opening the first valve 5 and the second valve 3 simultaneously; the first valve 5 and the second valve 3 are closed to realize the systematic sealing of the azotase activity measuring device, so that the influence of the air leakage of the device caused by the damage of the rubber plug 2 due to the fact that the needle tube with the needle head is used for pricking the rubber plug is reduced, and the experimental accuracy is further improved.
The syringe 1 for sucking gas should be exclusively used for gas.
The above examples are only preferred embodiments of the present invention, and it should be noted that: various modifications and changes may occur to those skilled in the art. All the equivalent replacements or improvements made according to the technical scheme and the utility model concept of the utility model are included in the spirit and principle of the utility model, and are included in the protection scope of the utility model.
Claims (1)
1. The azotase activity measuring device improved by using an acetylene reduction method is characterized by comprising a sealed reaction bottle (7), wherein an isobutyl rubber plug (6) is arranged at the mouth of the sealed reaction bottle (7), a gas sample inlet pipe (4) and a nutrient solution sample inlet pipe (9) are fixed on the isobutyl rubber plug (6), the gas sample inlet pipe (4) is sequentially connected with a first valve (5) and a second valve (3), and the other end of the gas sample inlet pipe (4) is communicated with an injector (1) through a rubber plug (2);
the nutrient solution sampling pipe (9) is sequentially provided with a third valve (10) and a multi-joint pipe valve (11); the multi-joint pipe valve (11) is connected with a nutrient solution bottle (12);
the sealed reaction bottle (7) is positioned in the porous heat-preservation type vibration plastic component (8), the component comprises a plurality of small holes, and the diameter of each small hole is the same as that of the sealed reaction bottle (7).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202021048209.2U CN212341119U (en) | 2020-06-10 | 2020-06-10 | Nitrogen fixation enzyme activity measuring device improved by acetylene reduction method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202021048209.2U CN212341119U (en) | 2020-06-10 | 2020-06-10 | Nitrogen fixation enzyme activity measuring device improved by acetylene reduction method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN212341119U true CN212341119U (en) | 2021-01-12 |
Family
ID=74077068
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202021048209.2U Active CN212341119U (en) | 2020-06-10 | 2020-06-10 | Nitrogen fixation enzyme activity measuring device improved by acetylene reduction method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN212341119U (en) |
-
2020
- 2020-06-10 CN CN202021048209.2U patent/CN212341119U/en active Active
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105973656A (en) | Soil respiration measurement culture and gas collection device and measurement method thereof | |
| CN102680545A (en) | Test instrument for detecting electrolyte item and total carbon dioxide | |
| CN212341119U (en) | Nitrogen fixation enzyme activity measuring device improved by acetylene reduction method | |
| CN102141535B (en) | Enzyme-injection glucose sensing on-line analyzer | |
| CN103792310B (en) | For the sampling device that water body release gas gas phase measures | |
| CN111562331A (en) | Nitrogen fixation enzyme activity determination device improved by acetylene reduction method and determination method thereof | |
| CN201234980Y (en) | Vacuum blood sampling tube | |
| CN105018343A (en) | Automated sample processing unit of original state microbial oil gas and hydrate exploration technology and automation workstation | |
| CN204575396U (en) | A kind of SF 6gas sample collecting device for air | |
| CN111366423B (en) | Device for collecting soil respiratory gas and gas collection method | |
| CN202849390U (en) | In-vitro batch culture gas production device capable of simulating rumen fermentation | |
| CN205374079U (en) | Bioprocess rapid sampling device | |
| CN115267026A (en) | High-pressure low-temperature phase balance measuring device | |
| CN1402009A (en) | Sampling diluting device for bio-reaction process in-line testing system | |
| CN213843015U (en) | Silicon water quality analyzer | |
| CN105018609A (en) | Automatic detection method of abundance of methane-oxidizing bacteria | |
| CN112322488A (en) | Bioreactor auxiliary device and operation method thereof | |
| CN214142375U (en) | Nucleic acid reagent reaction unit | |
| CN212321249U (en) | Tandem injection pump set structure for diluting trace liquid sample | |
| CN211856046U (en) | Device for collecting soil breathing gas | |
| CN210953984U (en) | Integrated automatic monitor for root secretion and respiration process | |
| CN117347457B (en) | Automatic calibration system and method for on-line detection of enzyme electrode of bioreactor | |
| CN211652292U (en) | Portable reticulocyte dyeing device | |
| CN2122386U (en) | Preparation device for carbon and oxygen stable isotope trace sample of carbonate | |
| US20180362916A1 (en) | Automated Bioreactor Sampling and Glucose monitoring System |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant |