CN111808774A - Recyclable oxygen indicator and preparation method and application thereof - Google Patents
Recyclable oxygen indicator and preparation method and application thereof Download PDFInfo
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- CN111808774A CN111808774A CN202010722787.8A CN202010722787A CN111808774A CN 111808774 A CN111808774 A CN 111808774A CN 202010722787 A CN202010722787 A CN 202010722787A CN 111808774 A CN111808774 A CN 111808774A
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Abstract
The invention discloses a recyclable oxygen indicator and a preparation method and application thereof.A strain Shewanella oneidensis MR-1 is inoculated in an LB culture medium and cultured overnight in a shaking table at 30 ℃ and 200rpm to obtain a seed solution; adding seed solution into decolorization medium containing sodium lactate and methylene blue to make OD600 value 0.05, and placing into anaerobic bottle; introducing nitrogen into the anaerobic bottle to replace air, covering the bottle plug, standing, and allowing the methylene blue dye to decolor; melting decolorization culture medium containing sodium lactate and 1% agar, and storing at 55 deg.C; and mixing the decolorized liquid with a decolorization culture medium containing 1% of agar according to the volume ratio of 1:1, and cooling to obtain the solid oxygen indicator. The invention utilizes an electrogenesis microorganism Shewanella oneidensis MR-1 to generate electrons by taking sodium lactate as an electron donor, reduces a bistable methylene blue dye solution to change from blue to colorless, then utilizes low-concentration agar to cure and prepare an oxygen indicator, and detects the anaerobic environment. The oxygen indicator prepared by the invention has higher sensitivity and can be recycled.
Description
Technical Field
The invention relates to a recyclable oxygen indicator and a preparation method and application thereof, belonging to the technical field of oxygen content measurement.
Background
Oxygen is an element that is not available in many biochemical reactions and is one of the major causes of deterioration of food and pharmaceutical agents. Meanwhile, the anaerobic environment is a production environment which needs to be strictly controlled in a plurality of industrial fields, for example, in sewage treatment plants, pharmaceutical factories, wineries, laboratories and other environments, the detection of oxygen in the anaerobic environment is necessary.
Oxygen indicators that have been used to date include electrochemical, optical and colorimetric oxygen indicators. The Clark electrode in electrochemical oxygen indicators is one of the earlier used oxygen sensors. However, the operation is complicated, the sensitivity is not high, the reaction time is long, and the like, so that the method is greatly limited in the practical application process. Among optical oxygen indicators, oxygen indication methods based on fluorescence development are of more interest. The indicator quenches the fluorescent substance by dynamic collision of oxygen and detects the quenching degree of fluorescence to characterize the content of oxygen. Compared with an electrochemical oxygen indicator, the optical oxygen indicator has no by-product and better stability, but the method needs special instrument equipment to detect the density of fluorescence, so that the application cost is higher. The colorimetric oxygen indicator overcomes the defects of an optical oxygen indicator, reflects the oxygen content through macroscopic color change, is quick, simple and convenient, and mainly comprises an oxygen complexing type, an oxidation reduction type and a light-driven oxidation reduction type at present. The oxygen complex type is a type in which a color change occurs by the participation of oxygen in a complex reaction, but has a disadvantage in that the color change is less noticeable. The redox type is an indication of the presence or absence of oxygen by a colour change in the reduced state of the dye's oxidation state agent. For example, indigo and thioindigo are used as irreversible oxygen indicators for dyes. However, these oxygen indicators are expensive and difficult to store. The light-driven redox type has a UV activated colorimetric oxygen indicator ink and an indicator of UVB activated titanium dioxide photocatalyst. Wu-Reinforcement et al use the reflected ultraviolet action of soapstone to introduce methylene blue and ascorbic acid into the interlayer structure of soapstone under the action of surfactant to prepare a novel oxygen indicator. However, in China, the oxygen indicator products are not abundant, and although part of the oxygen indicator is produced commercially, the oxygen indicator cannot be recycled basically, so that the resource waste is caused.
Disclosure of Invention
One of the objectives of the present invention is to provide a recyclable oxygen indicator, which can rapidly indicate the presence of oxygen in the environment and can be reused many times.
Another object of the present invention is to provide a method for preparing the above oxygen indicator which can be recycled.
It is a further object of the present invention to provide the use of such a recyclable oxygen indicator.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: a recyclable oxygen indicator comprising Shewanella oneidensis MR-1, sodium lactate, methylene blue and a solid decolorization medium consisting of: na (Na)2HPO4·12H2O 7.52g/L，NaH2PO4·2H2O7.13g/L，YE 2g/L，NH4Cl1g/L, NaCl 0.5g/L, agar 1%.
The method for regenerating the oxygen indicator comprises the following steps: and (4) standing the oxygen indicator after the oxygen oxidation color development for several hours again, and obtaining the regenerated oxygen indicator after the color is faded.
The invention also provides a preparation method of the recyclable oxygen indicator, which comprises the following steps:
step 1, inoculating a strain Shewanella oneidensis MR-1 to an LB culture medium, and carrying out shaking table overnight culture at 30 ℃ and 200rpm to obtain a seed solution;
step 2, adding sodium lactate and methylene blue into a decoloring culture mediumSeed liquid to OD600The value is 0.05, and then the mixture is filled into an anaerobic bottle;
step 3, introducing N into the anaerobic bottle2Covering a bottle stopper after 1min to ensure an anaerobic environment, standing, and decolorizing with methylene blue dye;
step 4, storing the decolorization medium containing sodium lactate and 1% agar at 55 ℃;
and 5, mixing the decolorized liquid obtained in the step 3 with the decolorized culture medium containing 1% of agar obtained in the step 4 according to the volume ratio of 1:1, and cooling to obtain the solid oxygen indicator.
In the step 2, the concentration of sodium lactate in the decoloring medium containing sodium lactate and methylene blue is 10mM, and the concentration of methylene blue is 20. mu.M.
In step 4, the concentration of sodium lactate in the decolorization medium containing sodium lactate and 1% agar was 10 mM.
The invention utilizes an electrogenic microorganism Shewanella oneidensis MR-1 (purchased from American culture Collection, number ATCC 700550) to generate electrons by taking sodium lactate as an electron donor, reduces a bistable methylene blue dye solution to be colorless from blue, and then utilizes low-concentration agar to cure to prepare an oxygen indicator and detect the anaerobic environment. In the using process, if the environment is strict anaerobic environment, the indicator is colorless; when oxygen exists in the environment, the methylene blue is oxidized into blue; after the use is finished, the cell is sealed again, the Shewanella oneidensis MR-1 consumes oxygen, and simultaneously, sodium lactate is used as an electron donor to reduce methylene blue to be colorless again, so that the blue can be recycled.
Compared with the prior art, the invention has the following beneficial effects:
1. the strain Shewanella oneidensis MR-1 used in the invention can grow in both anaerobic and aerobic environments, and the strain has high growth speed and is easy to culture.
2. The invention uses the bacterial strain to decolor the methylene blue, and the prepared oxygen indicator has higher sensitivity and obvious color change.
3. The oxygen indicator prepared by the invention can be recycled, the production cost is reduced, and the oxygen indicator is economic and environment-friendly.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
Example 1: preparation of oxygen indicators
Step 1, preparing seed liquid: inoculating the strain Shewanella oneidensis MR-1 into an LB culture medium, and performing shaking table overnight at the temperature of 30 ℃ and the speed of 200rpm to obtain a seed solution;
step 2, decoloring: subpackaging decolorization medium containing sodium lactate and methylene blue into anaerobic bottles, adding seed solution to make final OD600A value of 0.05; introducing N into the anaerobic bottle2Covering a bottle stopper after 1min to ensure an anaerobic environment; standing until the dye is decolorized; the decolorization medium comprises the following components: na (Na)2HPO4·12H2O 7.52g/L，NaH2PO4·2H2O 7.13g/L，YE 2g/L，NH4Cl1g/L and NaCl 0.5 g/L; wherein the concentration of sodium lactate is 10mM, and the concentration of methylene blue is 20 μ M;
step 3, preparing an anaerobic environment indicator: melting decolorization culture medium containing sodium lactate and 1% agar, and placing in 55 deg.C oven, and keeping the temperature, wherein the concentration of sodium lactate is 10 mM; mixing the decolorized liquid with melted decolorized culture medium containing agar at volume ratio of 1:1, cooling to obtain product, and storing in anaerobic bottle.
Example 2: use of oxygen indicators
Opening the bottle stopper of the anaerobic bottle, placing the oxygen indicator in an environment to be detected to observe the color change, if no oxygen exists in the detection environment, keeping the indicator colorless all the time, if contacting the oxygen in the detection environment, rapidly changing the upper layer of the agar into blue, and gradually deepening the blue part downwards along with the prolonging of the time of contacting the oxygen.
Covering a bottle stopper, standing for several hours, quickly consuming oxygen by the thallus, utilizing sodium lactate to generate electrons to reduce methylene blue again, and quickly fading the blue color to be recycled for more than 10 times.
Claims (6)
1. A recyclable oxygen indicator comprising Shewanellaone of ShewanellaoneThe traditional Chinese medicine composition comprises idensis MR-1, sodium lactate, methylene blue and a solid decolorizing medium, wherein the solid decolorizing medium comprises the following components: na (Na)2HPO4·12H2O 7.52g/L，NaH2PO4·2H2O 7.13g/L，YE 2g/L，NH4Cl1g/L, NaCl 0.5g/L, agar 1%.
2. A recyclable oxygen indicator according to claim 1 wherein the oxygen indicator is regenerated by: and (4) standing the oxygen indicator after the oxygen oxidation color development for several hours again, and obtaining the regenerated oxygen indicator after the color is faded.
3. A method of making a recyclable oxygen indicator as defined in claim 1 comprising the steps of:
step 1, inoculating a strain Shewanella oneidensis MR-1 to an LB culture medium, and carrying out shaking table overnight culture at 30 ℃ and 200rpm to obtain a seed solution;
step 2, adding seed liquid into decolorizing culture medium containing sodium lactate and methylene blue to make OD600The value is 0.05, and then the mixture is filled into an anaerobic bottle;
step 3, introducing nitrogen into the anaerobic bottle to replace air, covering a bottle plug to ensure an anaerobic environment, standing, and after the methylene blue dye is decolorized;
step 4, melting a decoloring culture medium containing sodium lactate and 1% agar, and preserving at 55 ℃;
and 5, mixing the decolorized liquid obtained in the step 3 with the decolorized culture medium containing 1% of agar obtained in the step 4 according to the volume ratio of 1:1, and cooling to obtain the solid oxygen indicator.
4. The method of claim 3, wherein in step 2, the concentration of sodium lactate in the decolorization medium containing sodium lactate and methylene blue is 10mM and the concentration of methylene blue is 20. mu.M.
5. The method of claim 3, wherein the concentration of sodium lactate in the decolorization medium containing sodium lactate and 1% agar in step 4 is 10 mM.
6. Use of the recyclable oxygen indicator of claim 1 in anaerobic environmental monitoring.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102315471A (en) * | 2010-07-07 | 2012-01-11 | 广东省微生物研究所 | Shewanella-decolorationis-based microbial fuel cell and using method thereof |
| CN104263672A (en) * | 2014-07-23 | 2015-01-07 | 常州市第一人民医院 | High-electricity-generation shewanella bacterium and application of high-electricity-generation shewanella bacterium |
| CN108841765A (en) * | 2018-07-24 | 2018-11-20 | 江苏师范大学 | Steady bacillus and its application in conversion sweet potato stalk production biological flocculant |
| CN109457001A (en) * | 2018-10-31 | 2019-03-12 | 浙江海洋大学 | A kind of extracellular polysaccharide and preparation method and application with decoloring ability |
| US20190284584A1 (en) * | 2018-03-15 | 2019-09-19 | Indian Oil Corporation Limited | Bio-assisted process for conversion of mixed volatile fatty acids to selective drop-in fuels |
-
2020
- 2020-07-24 CN CN202010722787.8A patent/CN111808774A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102315471A (en) * | 2010-07-07 | 2012-01-11 | 广东省微生物研究所 | Shewanella-decolorationis-based microbial fuel cell and using method thereof |
| CN104263672A (en) * | 2014-07-23 | 2015-01-07 | 常州市第一人民医院 | High-electricity-generation shewanella bacterium and application of high-electricity-generation shewanella bacterium |
| US20190284584A1 (en) * | 2018-03-15 | 2019-09-19 | Indian Oil Corporation Limited | Bio-assisted process for conversion of mixed volatile fatty acids to selective drop-in fuels |
| CN108841765A (en) * | 2018-07-24 | 2018-11-20 | 江苏师范大学 | Steady bacillus and its application in conversion sweet potato stalk production biological flocculant |
| CN109457001A (en) * | 2018-10-31 | 2019-03-12 | 浙江海洋大学 | A kind of extracellular polysaccharide and preparation method and application with decoloring ability |
Non-Patent Citations (6)
| Title |
|---|
| LIANG SHEN等: "Enhanced Treatment of Anionic and Cationic Dyes in Wastewater through Live Bacteria Encapsulation Using Graphene Hydrogel", 《INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH》 * |
| LIU XIAN-WEI等: "Phenothiazine derivative-accelerated microbial extracellular electron transfer in bioelectrochemical system", 《SCIENTIFIC REPORTS》 * |
| WEIJIELIU等: "Methylene blue enhances the anaerobic decolorization and detoxication of azo dye by Shewanella onediensis MR-1", 《BIOCHEMICAL ENGINEERING JOURNAL》 * |
| 刘亮等: "希瓦氏菌在印染废水脱色中的研究进展", 《环境科学与技术》 * |
| 孔祥义: "脱色希瓦氏菌铁还原性能及对偶氮染料 —一种非侵染性病害源的脱色研究", 《中国优秀硕士学位论文全文数据库》 * |
| 白蓉等主编: "《分析化学中的分析方法与应用研究》", 31 August 2018 * |
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