CN106978344A - The cultural method of Phenol-degrading Bacteria Strains - Google Patents
The cultural method of Phenol-degrading Bacteria Strains Download PDFInfo
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- CN106978344A CN106978344A CN201710247855.8A CN201710247855A CN106978344A CN 106978344 A CN106978344 A CN 106978344A CN 201710247855 A CN201710247855 A CN 201710247855A CN 106978344 A CN106978344 A CN 106978344A
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- phenol
- bacteria strains
- degrading bacteria
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/34—Organic compounds containing oxygen
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Abstract
The present invention relates to a kind of cultural method of Phenol-degrading Bacteria Strains, it is characterised in that comprises the following steps：(1) phenol degrading bacterium culture medium is prepared；(2) by Phenol-degrading Bacteria Strains activation in phenol degrading bacterium culture medium；(3) the Phenol-degrading Bacteria Strains solution diluted is taken to distinguish 200 microlitres etc. with liquid-transfering gun.It is an advantage of the invention that：The Phenol-degrading Bacteria Strains cultivated by the present invention, can effectively remove phenol material, and do not pollute the environment.
Description
Technical field
The present invention relates to a kind of cultural method of Phenol-degrading Bacteria Strains.
Background technology
It in current sewage, can all contain nitrite, but how clean effective removal to aldehydes matter in sewage,
And it is free from environmental pollution, still in conceptual phase.Pertinent literature is not found through retrieval.
The content of the invention
To overcome the defect of prior art, the present invention provides a kind of cultural method of Phenol-degrading Bacteria Strains, technology of the invention
Scheme is：
A kind of cultural method of Phenol-degrading Bacteria Strains, comprises the following steps：
(1) phenol degrading bacterium culture medium is prepared, specific preparation method is as follows：Ammonium sulfate 1g/L, dipotassium hydrogen phosphate 0.2g/L,
Magnesium sulfate 0.05g/L, phenol 0.5g/L, agar 18g/L, phenol degrading bacterium culture medium is prepared according to component ratio, and in microwave
Stove heat, heat time：2min, heating-up temperature：80 DEG C, untill agar melts completely, it is put into high-pressure sterilizing pot and is sterilized,
Then 121 DEG C of sterilising temp, time 30min pours into sterilized desiccation culture ware, each culture dish in Biohazard Safety Equipment
15ml is poured into, the culture dish of good culture medium is placed in safety cabinet to cooling, cool time：5h, chilling temperature：Room temperature 25
DEG C, refrigerator cold-storage can be put into after cooling standby；
(2) Phenol-degrading Bacteria Strains are activated in phenol degrading bacterium culture medium, then the Degradation of Phenol in superclean bench
Bacterium carries out flat board coating, while Phenol-degrading Bacteria Strains solution concentration is carried out into gradient dilution, 10-2 powers, 10 are diluted to respectively
- 4 powers ,-the 6 of 10 powers and-the 8 of 10 powers.
(3) take the Phenol-degrading Bacteria Strains solution diluted to distinguish 200 microlitres with liquid-transfering gun, be injected separately into the benzene in culture dish
It is with spreading rod that Phenol-degrading Bacteria Strains solution coating is uniform on Phenol degrading bacteria culture medium, and entered with marking pen in the bottom of culture dish
Rower is noted, and in the constant incubator for being then placed in 35 DEG C, is cultivated, after 1-2 days, observes the growing state of Phenol-degrading Bacteria Strains,
The Phenol-degrading Bacteria Strains of the different shape grown on culture medium, and reference number are marked respectively with marking pen, then to the benzene of mark
Phenol degrading bacteria carries out plate streaking；
(4) flat board pulled is put into constant incubator, 35 DEG C, cultivated 1-2 days, the life of Phenol-degrading Bacteria Strains on observation flat board
It is long, and dyeing microscopic examination is carried out to the Phenol-degrading Bacteria Strains grown, its morphological feature is recorded, the operation is repeated, until under microscope
Untill the Phenol-degrading Bacteria Strains form Economical Purification of observation, the Phenol-degrading Bacteria Strains of purifying are carried out to carry out in test tube slant culture medium
Inclined-plane line preservation.
Specific dilution process is in described step (2)：10ml distilled water is taken to be put into test tube with syringe, by distilled water
Sterilized, in Biohazard Safety Equipment, 100 microlitres of strain solution taken with liquid-transfering gun, is injected into the test tube of sterile distilled water,
Now, the strain solution concentration in the test tube is 10-2 powers, and strain solution is diluted to by same method respectively 10 for-4 times
Side, 10-6 powers and-the 8 of 10 powers.
Described step (3) plate streaking method is as follows：In Biohazard Safety Equipment, alcolhol burner is lighted, with the top of oese
The Phenol-degrading Bacteria Strains of mark sequence number are touched, are drawn on phenol degrading bacterium culture medium, the Phenol-degrading Bacteria Strains of each sequence number are on culture medium
4th area are drawn, oese is placed on alcolhol burner per standardized area and burnt, is repeated.
The preparation method of described step (4) test tube slant culture medium：Weigh peptone 10g/L, beef extract powder 5g/L, fine jade
Cosmetics 18g/L, sodium chloride 5g/L, are mixed according to proportioning, are heated after mixing in micro-wave oven, the heat time：1-2min, plus
Hot temperature：70-80 DEG C, untill agar powder melts completely, taken with 10ml syringe in 5ml injecting tubes, test tube is put into
2L beaker is wrapped up, and is then placed in high-pressure sterilizing pot and is sterilized, and is sterilized 121 DEG C, time 30min, after sterilizing, by test tube
Pendulum inclined-plane is taken out, the standard of putting on inclined-plane is the 2/3 of of length no more than whole test tube length of nutrient agar on inclined-plane,
Stand 8h, it is to be condensed after, be put into refrigerator cold-storage.
It is an advantage of the invention that：The Phenol-degrading Bacteria Strains cultivated by the present invention, can effectively remove aldehydes matter, and
Do not pollute the environment.
Embodiment
The invention will now be further described with reference to specific embodiments, advantages of the present invention and feature will be with description and
It is apparent.But these embodiments are only exemplary, do not constitute any limitation to the scope of the present invention.People in the art
Member to the details and form of technical solution of the present invention it should be understood that can enter without departing from the spirit and scope of the invention
Row modifications or substitutions, but these are changed and replacement is each fallen within protection scope of the present invention.
The present invention relates to a kind of cultural method of Phenol-degrading Bacteria Strains, comprise the following steps：
(1) phenol degrading bacterium culture medium is prepared, specific preparation method is as follows：Ammonium sulfate 1g/L, dipotassium hydrogen phosphate 0.2g/L,
Magnesium sulfate 0.05g/L, phenol 0.5g/L, agar 18g/L, phenol degrading bacterium culture medium is prepared according to component ratio, and in microwave
Stove heat, heat time：2min, heating-up temperature：80 DEG C, untill agar melts completely, it is put into high-pressure sterilizing pot and is sterilized,
Then 121 DEG C of sterilising temp, time 30min pours into sterilized desiccation culture ware, each culture dish in Biohazard Safety Equipment
15ml is poured into, the culture dish of good culture medium is placed in safety cabinet to cooling, cool time：5h, chilling temperature：Room temperature 25
DEG C, refrigerator cold-storage can be put into after cooling standby；
(2) Phenol-degrading Bacteria Strains are activated in phenol degrading bacterium culture medium, then the Degradation of Phenol in superclean bench
Bacterium carries out flat board coating, while Phenol-degrading Bacteria Strains solution concentration is carried out into gradient dilution, 10-2 powers, 10 are diluted to respectively
- 4 powers ,-the 6 of 10 powers and-the 8 of 10 powers.
(3) take the Phenol-degrading Bacteria Strains solution diluted to distinguish 200 microlitres with liquid-transfering gun, be injected separately into the benzene in culture dish
It is with spreading rod that Phenol-degrading Bacteria Strains solution coating is uniform on Phenol degrading bacteria culture medium, and entered with marking pen in the bottom of culture dish
Rower is noted, and in the constant incubator for being then placed in 35 DEG C, is cultivated, after 1-2 days, observes the growing state of Phenol-degrading Bacteria Strains,
The Phenol-degrading Bacteria Strains of the different shape grown on culture medium, and reference number are marked respectively with marking pen, then to the benzene of mark
Phenol degrading bacteria carries out plate streaking；
(4) flat board pulled is put into constant incubator, 35 DEG C, cultivated 1-2 days, the life of Phenol-degrading Bacteria Strains on observation flat board
It is long, and dyeing microscopic examination is carried out to the Phenol-degrading Bacteria Strains grown, its morphological feature is recorded, the operation is repeated, until under microscope
Untill the Phenol-degrading Bacteria Strains form Economical Purification of observation, the Phenol-degrading Bacteria Strains of purifying are carried out to carry out in test tube slant culture medium
Inclined-plane line preservation.
Specific dilution process is in described step (2)：10ml distilled water is taken to be put into test tube with syringe, by distilled water
Sterilized, in Biohazard Safety Equipment, 100 microlitres of strain solution taken with liquid-transfering gun, is injected into the test tube of sterile distilled water,
Now, the strain solution concentration in the test tube is 10-2 powers, and strain solution is diluted to by same method respectively 10 for-4 times
Side, 10-6 powers and-the 8 of 10 powers.
Described step (3) plate streaking method is as follows：In Biohazard Safety Equipment, alcolhol burner is lighted, with the top of oese
The Phenol-degrading Bacteria Strains of mark sequence number are touched, are drawn on phenol degrading bacterium culture medium, the Phenol-degrading Bacteria Strains of each sequence number are on culture medium
4th area are drawn, oese is placed on alcolhol burner per standardized area and burnt, is repeated.
The preparation method of described step (4) test tube slant culture medium：Weigh peptone 10g/L, beef extract powder 5g/L, fine jade
Cosmetics 18g/L, sodium chloride 5g/L, are mixed according to proportioning, are heated after mixing in micro-wave oven, the heat time：1-2min, plus
Hot temperature：70-80 DEG C, untill agar powder melts completely, taken with 10ml syringe in 5ml injecting tubes, test tube is put into
2L beaker is wrapped up, and is then placed in high-pressure sterilizing pot and is sterilized, and is sterilized 121 DEG C, time 30min, after sterilizing, by test tube
Pendulum inclined-plane is taken out, the standard of putting on inclined-plane is the 2/3 of of length no more than whole test tube length of nutrient agar on inclined-plane,
Stand 8h, it is to be condensed after, be put into refrigerator cold-storage.
Embodiment：The sewage with aldehydes matter is taken, the wherein content of phenol type substances is 0.138g/L, adds the present invention
Phenol-degrading Bacteria Strains after purification, the content of phenol type substances is 0.069g/L, as a result shows, the clearance of aldehydes matter is reached
50%.
Claims (4)
1. a kind of cultural method of Phenol-degrading Bacteria Strains, it is characterised in that comprise the following steps：
(1) phenol degrading bacterium culture medium is prepared, specific preparation method is as follows：Ammonium sulfate 1g/L, dipotassium hydrogen phosphate 0.2g/L, sulfuric acid
Magnesium 0.05g/L, phenol 0.5g/L, agar 18g/L, prepare phenol degrading bacterium culture medium, and add in micro-wave oven according to component ratio
Heat, heat time：2min, heating-up temperature：80 DEG C, untill agar melts completely, it is put into high-pressure sterilizing pot and is sterilized, sterilizes
Then 121 DEG C of temperature, time 30min pours into sterilized desiccation culture ware, each culture dish is poured into Biohazard Safety Equipment
15ml, the culture dish of good culture medium is placed in safety cabinet to cooling, cool time：5h, chilling temperature：25 DEG C of room temperature, it is cold
But refrigerator cold-storage can be put into after standby；
(2) by Phenol-degrading Bacteria Strains activation in phenol degrading bacterium culture medium, then Degradation of Phenol bacterium enters in superclean bench
Row flat board is coated with, while Phenol-degrading Bacteria Strains solution concentration is carried out into gradient dilution, 10-2 powers ,-4 times of 10 are diluted to respectively
Side, 10-6 powers and-the 8 of 10 powers.
(3) take the Phenol-degrading Bacteria Strains solution diluted to distinguish 200 microlitres with liquid-transfering gun, be injected separately into the phenol drop in culture dish
Solve on bacterium culture medium, it is with spreading rod that Phenol-degrading Bacteria Strains solution coating is uniform, and enter rower in the bottom of culture dish with marking pen
In note, the constant incubator for being then placed in 35 DEG C, cultivated, after 1-2 days, the growing state of Phenol-degrading Bacteria Strains is observed, with note
Number pen marks the Phenol-degrading Bacteria Strains of the different shape grown on culture medium, and reference number respectively, and then the phenol of mark is dropped
Solve bacterium and carry out plate streaking；
(4) flat board pulled is put into constant incubator, 35 DEG C, cultivated 1-2 days, the growth of Phenol-degrading Bacteria Strains on observation flat board,
And dyeing microscopic examination is carried out to the Phenol-degrading Bacteria Strains grown, its morphological feature is recorded, the operation is repeated, until micro- Microscopic observation
Phenol-degrading Bacteria Strains form Economical Purification untill, the Phenol-degrading Bacteria Strains of purifying are carried out to carry out inclined-plane in test tube slant culture medium
Line preservation.
2. the cultural method of Phenol-degrading Bacteria Strains according to claim 1, it is characterised in that specific in described step (2)
Dilution process is：Take 10ml distilled water to be put into test tube with syringe, distilled water is sterilized, in Biohazard Safety Equipment, use
Liquid-transfering gun takes 100 microlitres of strain solution, is injected into the test tube of sterile distilled water, now, and the strain solution in the test tube is dense
Spend-2 powers for 10, same method, strain solution is diluted to respectively 10-4 powers ,-the 6 of 10 powers and-the 8 of 10 times
Side.
3. the cultural method of Phenol-degrading Bacteria Strains according to claim 1, it is characterised in that described step (3) flat board is drawn
Line method is as follows：In Biohazard Safety Equipment, alcolhol burner is lighted, with the Phenol-degrading Bacteria Strains that mark sequence number is touched at the top of oese, is drawn
On phenol degrading bacterium culture medium, oese is placed on by the Phenol-degrading Bacteria Strains of each sequence number in the areas of culture medium Shang Hua tetra- per standardized area
Burn, repeat on alcolhol burner.
4. the cultural method of Phenol-degrading Bacteria Strains according to claim 1, it is characterised in that described step (4) test tube is oblique
The preparation method of face culture medium：Weigh peptone 10g/L, beef extract powder 5g/L, agar powder 18g/L, sodium chloride 5g/L, according to
Than being mixed, heated after mixing in micro-wave oven, the heat time：1-2min, heating-up temperature：It is 70-80 DEG C, complete to agar powder
Untill thawing, taken with 10ml syringe in 5ml injecting tubes, the beaker that test tube is put into 2L is wrapped up, and is then placed in height
Pressure autoclave is sterilized, and is sterilized 121 DEG C, time 30min, after sterilizing, and test tube is taken out into pendulum inclined-plane, and the standard of putting on inclined-plane is
The 2/3 of of length no more than whole test tube length of nutrient agar on inclined-plane, stand 8h, it is to be condensed after, be put into refrigerator cold
Tibetan.
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| CN201710247855.8A CN106978344A (en) | 2017-04-17 | 2017-04-17 | The cultural method of Phenol-degrading Bacteria Strains |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659927A (en) * | 2009-07-29 | 2010-03-03 | 南京师范大学 | Trichosporon montevideense and application thereof in degradation of phenol |
| CN101857847A (en) * | 2010-05-06 | 2010-10-13 | 合肥工业大学 | Pseudomonas aeruginosa strain separating, purifying and domesticating method and use |
| CN105907688A (en) * | 2016-06-27 | 2016-08-31 | 南京工业大学 | Strain for degrading phenol compounds and application of strain |
| CN106119151A (en) * | 2016-06-21 | 2016-11-16 | 盐城工学院 | Bacterial strain and method for screening and separating thereof for degraded 2,4,6 trichlorophenol, 2,4,6,-Ts |
-
2017
- 2017-04-17 CN CN201710247855.8A patent/CN106978344A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659927A (en) * | 2009-07-29 | 2010-03-03 | 南京师范大学 | Trichosporon montevideense and application thereof in degradation of phenol |
| CN101857847A (en) * | 2010-05-06 | 2010-10-13 | 合肥工业大学 | Pseudomonas aeruginosa strain separating, purifying and domesticating method and use |
| CN106119151A (en) * | 2016-06-21 | 2016-11-16 | 盐城工学院 | Bacterial strain and method for screening and separating thereof for degraded 2,4,6 trichlorophenol, 2,4,6,-Ts |
| CN105907688A (en) * | 2016-06-27 | 2016-08-31 | 南京工业大学 | Strain for degrading phenol compounds and application of strain |
Non-Patent Citations (7)
| Title |
|---|
| 曹澍泽: "《兽医微生物学及免疫学技术》", 31 May 1992, 北京农业大学出版社 * |
| 李鲜珠等: "苯酚降解菌筛选及降解特性研究", 《水资源保护》 * |
| 葛启隆等: "一株苯酚降解菌的分离鉴定及响应面法优化其固定化", 《中国环境科学》 * |
| 贺强礼等: "一株苯酚降解菌的筛选鉴定及响应面法优化其降解", 《环境科学学报》 * |
| 颜家保等: "苯酚降解菌的分离及其降解特性研究", 《武汉科技大学学报》 * |
| 黄亚东等: "《微生物实验技术》", 31 January 2013, 中国轻工业出版社 * |
| 龚斌等: "一株高效苯酚降解菌的分离 、鉴定及降解特性的研究", 《环境科学学报》 * |
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Application publication date: 20170725 |