CN101307353A - Screening method of desulfurizing denitrification autotrophic bacteria - Google Patents
Screening method of desulfurizing denitrification autotrophic bacteria Download PDFInfo
- Publication number
- CN101307353A CN101307353A CNA2008100644681A CN200810064468A CN101307353A CN 101307353 A CN101307353 A CN 101307353A CN A2008100644681 A CNA2008100644681 A CN A2008100644681A CN 200810064468 A CN200810064468 A CN 200810064468A CN 101307353 A CN101307353 A CN 101307353A
- Authority
- CN
- China
- Prior art keywords
- autotrophic bacteria
- desulfurizing denitrification
- denitrification autotrophic
- substratum
- nutrient medium
- 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.)
- Pending
Links
Images
Abstract
A method for screening desulfurization-denitrogenation autotrophic bacteria relates to a method for screening autotrophic bacteria. The method solves the problems that the prior method for separating the desulfurization-denitrogenation autotrophic bacteria is complex and an obtained monocolony has poor growth. The method comprises the following steps that: (1) sample bacteria suspension is taken to carry out separated amplification culture; (2) a monocolony is cultured by an anaerobic interlayer; (3) the monocolony is selected to carry out amplification culture; (4) bacteria liquid obtained after amplification culture is selected and inoculated in solid separation culture till the monocolony grows up, and then the monocolony is selected and transferred to a liquid medium; and (5) the step (2), the step (3) and the step (4) are repeated till the monocolony is separated, and the separated monocolony is the desulfurization-denitrogenation autotrophic bacteria. The desulfurization-denitrogenation autotrophic bacteria use simple method and have sound growth of monocolony.
Description
Technical field
The present invention relates to a kind of screening method of autotrophic bacteria.
Background technology
Biological desulfurization process is because of the clearance height, need not to add chemical agent, can generate advantages such as elemental sulfur recovery and receive publicity day by day, wherein Thiobacillus (Thiobacillus) is the microorganism that a class has the desulfurizing denitrification function, they can obtain energy by oxidation of sulfur compounds, simultaneously nitrate can also be reduced to nitrogen as electron acceptor(EA) with it, do not need to add organic carbon source in the whole desulfurizing denitrification process, and can be by the suitable ecological condition of control, obtain maximized elemental sulfur transformation efficiency, realizing the recovery of elemental sulfur, is the significant engineering bacteria of a class.
At present; separating the desulfurizing denitrification autotrophic bacteria bacterial strain all is to adopt plate streak; but because the growth of desulfurizing denitrification autotrophic bacteria is very slow; time in epoch is longer; difficultly on substratum, obtain the good single bacterium colony of growth, and when adopting plate streak, the storage condition of substratum need keep anaerobic condition; so also make to become complicated more, increased the difficulty that obtains pure culture in the process of separating pure bacterium.
Summary of the invention
The present invention seeks to have method complexity, the bad problem of gained list colony growth of separating desulfurizing denitrification autotrophic bacteria now, and a kind of screening method of desulfurizing denitrification autotrophic bacteria is provided in order to solve.
A kind of screening method of desulfurizing denitrification autotrophic bacteria is realized according to the following steps: one, the sample thief bacterial suspension inoculation separates enlarged culturing under 26～30 ℃, the condition of 140～180r/min in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium, and incubation period is 14d; Two, get the bacterial suspension inoculation that separates after the enlarged culturing on solid separation culture medium, water and cover substratum and make anaerobism interlayer substratum, constant temperature culture bubble occurs to the substratum interlayer under 26～30 ℃ condition; Three, the single bacterium colony in the picking bubble is transferred in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium, and constant temperature enlarged culturing to substratum becomes turbid under 26～30 ℃ condition; Four, the bacterium liquid in the picking substratum is inoculated on the solid separation culture medium, and constant temperature culture is to there being single bacterium colony to grow up under 26～30 ℃ condition, and picking list bacterium colony is transferred in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium then; Five, repeating step two, three and four is desulfurizing denitrification autotrophic bacteria to isolating single bacterium colony; Wherein used desulfurizing denitrification autotrophic bacteria liquid nutrient medium is identical in the step 1, three and four, and every liter of desulfurizing denitrification autotrophic bacteria liquid nutrient medium all is the Na by 5.0g
2S
2O
35H
2The NH of O, 1.0g
4The Na of Cl, 2.0g
2HPO
4, 0.8g MgSO
47H
2The NaHCO of O, 3.0g
3, 3.5g KNO
3, the EDTA of 0.5mg, the ZnSO of 22.0ug
4, 55.0ug CaCl
2, 50.6ug MnCl
24H
2(the NH of O, 11.0ug
4)
6MoO
244H
2The CuSO of O, 15.7ug
45H
2The CoCl of O, 16.1ug
26H
2The VITAMIN liquid of O, 1mL and the distilled water of surplus are formed, and desulfurizing denitrification autotrophic bacteria liquid nutrient medium pH value is 7.0; Used solid separation culture medium is identical in the step 2 and four, and every liter of desulfurizing denitrification autotrophic bacteria solid body substratum all is the Na by 5.0g
2S
2O
35H
2The NH of O, 1.0g
4The Na of Cl, 2.0g
2HPO
4, 0.8g MgSO
47H
2The NaHCO of O, 3.0g
3, 3.5g KNO
3, the EDTA of 0.5mg, the ZnSO of 22.0ug
4, 55.0ug CaCl
2, 50.6ug MnCl
24H
2(the NH of O, 11.0ug
4)
6MoO
244H
2The CuSO of O, 15.7ug
45H
2The CoCl of O, 16.1ug
26H
2The distilled water of the VITAMIN liquid of O, 1mL, the agar of 20g and surplus is formed, and desulfurizing denitrification autotrophic bacteria liquid nutrient medium pH value is 7.0.
The present invention adopts anaerobism sandwiching screening desulfurizing denitrification autotrophic bacteria, the anaerobism interlayer can keep the anoxic condition of micro amount of oxygen, be more suitable for like this that desulfurizing denitrification autotrophic bacteria is faster and better on substratum to grow single bacterium colony, the anaerobism sandwiching need not deposited under the strictly anaerobic condition, being put into indoor environment gets final product, so also simplified the experimental implementation program, increased the suitability of separation method, on the interlayer substratum, can also see the aerogenesis situation of bacterium colony, therefore can judge which single bacterium colony is to need isolating desulfurizing denitrification autotrophic bacteria to fall, and has also improved the efficient of bacterial screening simultaneously.
Description of drawings
Fig. 1 is the transmission electron microscope picture of screening gained desulfurizing denitrification autotrophic bacteria in the embodiment five.
Embodiment
Embodiment one: the screening method of a kind of desulfurizing denitrification autotrophic bacteria of present embodiment is realized according to the following steps: one, the sample thief bacterial suspension inoculation is in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium, separate enlarged culturing under 26～30 ℃, the condition of 140～180r/min, incubation period is 14d; Two, get the bacterial suspension inoculation that separates after the enlarged culturing on solid separation culture medium, water and cover substratum and make anaerobism interlayer substratum, constant temperature culture bubble occurs to the substratum interlayer under 26～30 ℃ condition; Three, the single bacterium colony in the picking bubble is transferred in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium, and constant temperature enlarged culturing to substratum becomes turbid under 26～30 ℃ condition; Four, the bacterium liquid in the picking substratum is inoculated on the solid separation culture medium, and constant temperature culture is to there being single bacterium colony to grow up under 26～30 ℃ condition, and picking list bacterium colony is transferred in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium then; Five, repeating step two, three and four is desulfurizing denitrification autotrophic bacteria to isolating single bacterium colony; Wherein used desulfurizing denitrification autotrophic bacteria liquid nutrient medium is identical in the step 1, three and four, and every liter of desulfurizing denitrification autotrophic bacteria liquid nutrient medium all is the Na by 5.0g
2S
2O
35H
2The NH of O, 1.0g
4The Na of Cl, 2.0g
2HPO
4, 0.8g MgSO
47H
2The NaHCO of O, 3.0g
3, 3.5g KNO
3, the EDTA of 0.5mg, the ZnSO of 22.0ug
4, 55.0ug CaCl
2, 50.6ug MnCl
24H
2(the NH of O, 11.0ug
4)
6MoO
244H
2The CuSO of O, 15.7ug
45H
2The CoCl of O, 16.1ug
26H
2The VITAMIN liquid of O, 1mL and the distilled water of surplus are formed, and desulfurizing denitrification autotrophic bacteria liquid nutrient medium pH value is 7.0; Used solid separation culture medium is identical in the step 2 and four, and every liter of desulfurizing denitrification autotrophic bacteria solid body substratum all is the Na by 5.0g
2S
2O
35H
2The NH of O, 1.0g
4The Na of Cl, 2.0g
2HPO
4, 0.8g MgSO
47H
2The NaHCO of O, 3.0g
3, 3.5g KNO
3, the EDTA of 0.5mg, the ZnSO of 22.0ug
4, 55.0ug CaCl
2, 50.6ug MnCl
24H
2(the NH of O, 11.0ug
4)
6MoO
244H
2The CuSO of O, 15.7ug
45H
2The CoCl of O, 16.1ug
26H
2The distilled water of the VITAMIN liquid of O, 1mL, the agar of 20g and surplus is formed, and desulfurizing denitrification autotrophic bacteria liquid nutrient medium pH value is 7.0.
Embodiment two: what present embodiment and embodiment one were different is sample bacteria suspension preparation in the step 1: will gather the Erlenmeyer flask that zoogloea or soil particle are put into sterilization, add the sterilized water dilution, add 20～25 of granulated glass spherees, with 220r/min vibration 24h.Other step and parameter are identical with embodiment one.
Embodiment three: present embodiment and embodiment one are different is to separate enlarged culturing in the step 1 under 28 ℃, the condition of 160r/min.Other step and parameter are identical with embodiment one.
Embodiment four: present embodiment and embodiment one are different is to water in the step 2 that to cover temperature be 45～50 ℃.Other step and parameter are identical with embodiment one.
Embodiment five: the screening method of a kind of desulfurizing denitrification autotrophic bacteria of present embodiment is realized according to the following steps: one, the sample thief bacterial suspension inoculation is in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium, separate enlarged culturing under 28 ℃, the condition of 160r/min, incubation period is 14d; Two, get the bacterial suspension inoculation that separates after the enlarged culturing on solid separation culture medium, water and cover substratum and make anaerobism interlayer substratum, constant temperature culture bubble occurs to the substratum interlayer under 28 ℃ condition; Three, the single bacterium colony in the picking bubble is transferred in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium, and constant temperature enlarged culturing to substratum becomes turbid under 28 ℃ condition; Four, the bacterium liquid in the picking substratum is inoculated on the solid separation culture medium, and constant temperature culture is to there being single bacterium colony to grow up under 28 ℃ condition, and picking list bacterium colony is transferred in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium then; Five, repeating step two, three and four is desulfurizing denitrification autotrophic bacteria to isolating single bacterium colony; Wherein used desulfurizing denitrification autotrophic bacteria liquid nutrient medium is identical in the step 1, three and four, and every liter of desulfurizing denitrification autotrophic bacteria liquid nutrient medium all is the Na by 5.0g
2S
2O
35H
2The NH of O, 1.0g
4The Na of Cl, 2.0g
2HPO
4, 0.8g MgSO
47H
2The NaHCO of O, 3.0g
3, 3.5g KNO
3, the EDTA of 0.5mg, the ZnSO of 22.0ug
4, 55.0ug CaCl
2, 50.6ug MnCl
24H
2(the NH of O, 11.0ug
4)
6MoO
244H
2The CuSO of O, 15.7ug
45H
2The CoCl of O, 16.1ug
26H
2The VITAMIN liquid of O, 1mL and the distilled water of surplus are formed, and desulfurizing denitrification autotrophic bacteria liquid nutrient medium pH value is 7.0; Used solid separation culture medium is identical in the step 2 and four, and every liter of desulfurizing denitrification autotrophic bacteria solid body substratum all is the Na by 5.0g
2S
2O
35H
2The NH of O, 1.0g
4The Na of Cl, 2.0g
2HPO
4, 0.8g MgSO
47H
2The NaHCO of O, 3.0g
3, 3.5g KNO
3, the EDTA of 0.5mg, the ZnSO of 22.0ug
4, 55.0ug CaCl
2, 50.6ug MnCl
24H
2(the NH of O, 11.0ug
4)
6MoO
244H
2The CuSO of O, 15.7ug
45H
2The CoCl of O, 16.1ug
26H
2The distilled water of the VITAMIN liquid of O, 1mL, the agar of 20g and surplus is formed, and desulfurizing denitrification autotrophic bacteria liquid nutrient medium pH value is 7.0.
The desulfurizing denitrification autotrophic bacteria that the present embodiment screening obtains, Gram-negative, the form of bacterial strain is by shown in Figure 1, it is shaft-like that cell is, and single, paired or short chain shape is arranged, and has single polar flagella, no gemma, the thalline size is (0.3～0.5 μ m) * (10～15 μ m), to the evaluation of its physio-biochemical characteristics of carrying out and desulfurizing denitrification ability thereof, determines that the bacterial strain that screening obtains is a desulfurizing denitrification autotrophic bacteria.
Claims (4)
1, a kind of screening method of desulfurizing denitrification autotrophic bacteria, the screening method that it is characterized in that desulfurizing denitrification autotrophic bacteria is realized according to the following steps: one, the sample thief bacterial suspension inoculation is in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium, separate enlarged culturing under 26～30 ℃, the condition of 140～180r/min, incubation period is 14d; Two, get the bacterial suspension inoculation that separates after the enlarged culturing on solid separation culture medium, water and cover substratum and make anaerobism interlayer substratum, constant temperature culture bubble occurs to the substratum interlayer under 26～30 ℃ condition; Three, the single bacterium colony in the picking bubble is transferred in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium, and constant temperature enlarged culturing to substratum becomes turbid under 26～30 ℃ condition; Four, the bacterium liquid in the picking substratum is inoculated on the solid separation culture medium, and constant temperature culture is to there being single bacterium colony to grow up under 26～30 ℃ condition, and picking list bacterium colony is transferred in the desulfurizing denitrification autotrophic bacteria liquid nutrient medium then; Five, repeating step two, three and four is desulfurizing denitrification autotrophic bacteria to isolating single bacterium colony; Wherein used desulfurizing denitrification autotrophic bacteria liquid nutrient medium is identical in the step 1, three and four, and every liter of desulfurizing denitrification autotrophic bacteria liquid nutrient medium all is the Na by 5.0g
2S
2O
35H
2The NH of O, 1.0g
4The Na of Cl, 2.0g
2HPO
4, 0.8g MgSO
47H
2The NaHCO of O, 3.0g
3, 3.5g KNO
3, the EDTA of 0.5mg, the ZnSO of 22.0ug
4, 55.0ug CaCl
2, 50.6ug MnCl
24H
2(the NH of O, 11.0ug
4)
6MoO
244H
2The CuSO of O, 15.7ug
45H
2The CoCl of O, 16.1ug
26H
2The VITAMIN liquid of O, 1mL and the distilled water of surplus are formed, and desulfurizing denitrification autotrophic bacteria liquid nutrient medium pH value is 7.0; Used solid separation culture medium is identical in the step 2 and four, and every liter of desulfurizing denitrification autotrophic bacteria solid body substratum all is the Na by 5.0g
2S
2O
35H
2The NH of O, 1.0g
4The Na of Cl, 2.0g
2HPO
4, 0.8g MgSO
47H
2The NaHCO of O, 3.0g
3, 3.5g KNO
3, the EDTA of 0.5mg, the ZnSO of 22.0ug
4, 55.0ug CaCl
2, 50.6ug MnCl
24H
2(the NH of O, 11.0ug
4)
6MoO
244H
2The CuSO of O, 15.7ug
45H
2The CoCl of O, 16.1ug
26H
2The distilled water of the VITAMIN liquid of O, 1mL, the agar of 20g and surplus is formed, and desulfurizing denitrification autotrophic bacteria liquid nutrient medium pH value is 7.0.
2, the screening method of a kind of desulfurizing denitrification autotrophic bacteria according to claim 1, it is characterized in that the preparation of sample bacteria suspension in the step 1: the Erlenmeyer flask that the zoogloea gathered or soil particle are put into sterilization, add the sterilized water dilution, add 20～25 of granulated glass spherees, with 220r/min vibration 24h.
3, the screening method of a kind of desulfurizing denitrification autotrophic bacteria according to claim 1 is characterized in that in the step 1 separating enlarged culturing under 28 ℃, the condition of 160r/min.
4, the screening method of a kind of desulfurizing denitrification autotrophic bacteria according to claim 1 is characterized in that watering in the step 2 that to cover temperature be 45～50 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100644681A CN101307353A (en) | 2008-05-09 | 2008-05-09 | Screening method of desulfurizing denitrification autotrophic bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100644681A CN101307353A (en) | 2008-05-09 | 2008-05-09 | Screening method of desulfurizing denitrification autotrophic bacteria |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101307353A true CN101307353A (en) | 2008-11-19 |
Family
ID=40124031
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008100644681A Pending CN101307353A (en) | 2008-05-09 | 2008-05-09 | Screening method of desulfurizing denitrification autotrophic bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101307353A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103184152A (en) * | 2011-12-30 | 2013-07-03 | 中国石油天然气股份有限公司 | Method for screening natural gas biological desulphurization flora |
| CN104450592A (en) * | 2014-12-31 | 2015-03-25 | 哈尔滨工业大学 | Method for separating denitrification desulfurizing bacteria based on biodiversity information |
| CN106318871A (en) * | 2016-10-20 | 2017-01-11 | 中国石油化工股份有限公司 | Strain screening method with efficient desulfurization and denitrification activities simultaneously |
-
2008
- 2008-05-09 CN CNA2008100644681A patent/CN101307353A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103184152A (en) * | 2011-12-30 | 2013-07-03 | 中国石油天然气股份有限公司 | Method for screening natural gas biological desulphurization flora |
| CN103184152B (en) * | 2011-12-30 | 2014-12-24 | 中国石油天然气股份有限公司 | Method for screening natural gas biological desulphurization flora |
| CN104450592A (en) * | 2014-12-31 | 2015-03-25 | 哈尔滨工业大学 | Method for separating denitrification desulfurizing bacteria based on biodiversity information |
| CN106318871A (en) * | 2016-10-20 | 2017-01-11 | 中国石油化工股份有限公司 | Strain screening method with efficient desulfurization and denitrification activities simultaneously |
| CN106318871B (en) * | 2016-10-20 | 2019-09-27 | 中国石油化工股份有限公司 | A kind of bacterial strain screening method with high-efficiency desulfurization denitrification activity simultaneously |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR20210124232A (en) | Bacillus tianshenni strain, its application in the field of microbial preparation and heavy metal recovery | |
| Lutzu et al. | Interactions of microalgae and other microorganisms for enhanced production of high-value compounds | |
| Mutanda et al. | The utilization of post-chlorinated municipal domestic wastewater for biomass and lipid production by Chlorella spp. under batch conditions | |
| WO2017177773A1 (en) | Highly efficient aerobic phosphorus-removing bacteria capable of synthesizing nanoparticles by microbial self-assembly using waste water | |
| CN102643751A (en) | Method for quickly separating and purifying chlorella | |
| CA2723655A1 (en) | Biosynthesis of nanoparticles | |
| CN103923867B (en) | Mixed bacterial microbial preparation and in process containing the application in nitric nitrogen waste water | |
| CN104450592A (en) | Method for separating denitrification desulfurizing bacteria based on biodiversity information | |
| CN107177530B (en) | Novel efficient domestic sewage denitrifying bacterium and application thereof | |
| CN101955904B (en) | Algicidal bacteria separation method in natural water environment | |
| CN103255123B (en) | Method for mycelium pellet to form mixed mycelium pellet by adsorbing photosynthetic bacteria | |
| CN101307353A (en) | Screening method of desulfurizing denitrification autotrophic bacteria | |
| Abedi et al. | Decoupling a novel Trichormus variabilis-Synechocystis sp. interaction to boost phycoremediation | |
| Caldwell et al. | An ecological study of the sulfur-oxidizing bacteria from the littoral zone of a Michigan lake and a sulfur spring in Florida | |
| CN102827786A (en) | Lysinibacillus sp. for degrading S<2-> in petrochemical wastewater | |
| CN105802894B (en) | One pyridine degradation bacterium strain and its application in wastewater treatment containing pyridine | |
| CN115433694A (en) | Application of radiation-resistant methylobacterium L321 in degrading phthalate and promoting growth | |
| Halim et al. | Immobilized Microalgae using Alginate for Wastewater Treatment. | |
| Kuroda et al. | Biomass production and nutrient removal through cultivation of Euglena gracilis in domestic wastewater | |
| Kabir et al. | Nutrients utilization and biomass production by microalgae culture development in wastewater | |
| NL2011472C2 (en) | Storage compound production by phototrophic diatoms. | |
| CN108410758B (en) | Triazole degrading bacteria and application thereof in treatment of wastewater containing triazole | |
| CN107699511B (en) | Thermophilic thiobacillus and culture medium and culture method thereof | |
| CN115161229B (en) | Strain with sulfur reduction and iron reduction capabilities | |
| CN105462869B (en) | The preparation method of clostridium bifermentans and application thereof and nano metal sulphide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Open date: 20081119 |