CN106497837A - Gold orange II degradation bacterias AO7 2 and its microbial inoculum of production - Google Patents
Gold orange II degradation bacterias AO7 2 and its microbial inoculum of production Download PDFInfo
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Abstract
The invention discloses the microbial inoculum of gold orange II degradation bacterias AO7 2 and its production, the bacterial strain of degradation bacteria AO7 2 is Arthrobacter protophormiae, is preserved in China typical culture collection center, address within 19th in September in 2016：Wuhan University of Wuhan, China city, deposit number：CCTCC M2016502, Main Biological：Microorganism is in spherical, dispersed arrangement, and bacterium colony is in faint yellow, circle, moistening is glossy, opaque, surface elevation, neat in edge, without pod membrane, without gemma, Gram's staining is the positive to bacterium, bacterium is bred by fragmentation, and the bacterium is heterotrophic bacterium, in necessary for growth oxygen, sunlight is not needed, catalase reacting positive, oxydase reaction are negative.Optimum growth temperature is 30 C, and environmental optima pH is 8.0, and the residual quantity of azo dyes gold orange II can be made to reduce by more than 90%, microbial inoculum production and application low cost, easy to use, removal effect is suitable for the pollution control of related dye wastewater biological intensive treatment and water body or soil.
Description
Technical field
The invention belongs to biological technical field, and in particular to the microbial inoculum of gold orange II degradation bacterias AO7-2 and its production, it is to utilize
The method degraded azo dyes gold orange II of microorganism, it is adaptable to the control of environmental pollution.
Background technology
Azo dyes（Azo group two ends connect a class compound of aryl）It is that fabric clothing is applied in dyeing and printing process
Widest class synthetic dyestuffs, the dyeing and stamp for multiple natural and synthetic fibers, are also used for paint, plastics, rubber
Deng coloring.Under specific condition, it can decompose more than 20 kinds of carcinogenic aromatic amine of generation, through the DNA that activation changes human body
Structure causes pathology and induction cancer.
The common feature of azo-compound is to contain azo bond "-N=N- " in molecule.In general, azo-compound point
Containing 1-3 "-N=N- " in son, stupid or phenyl on key, is connected with, benzene or phenyl are connected with-Cl ,-NH again2、-CH3、-SO3、-NO2And-
The groups such as OH.
It is big that what azo dyes polluted is mainly characterized by contaminant capacity, although the dye component concentration remained in azo dye wastewater
Very low, but enter water body and the light transmittance of water body can be caused to reduce, destroy water ecosystem.These dye structures are stable, tool
There are alkali resistant, the characteristic such as antiacid, antimicrobial, anti-light, can be detained for a long time in the environment, therefore there is potential health
Harm.Where producing and using dyestuff, the important pollutant of underground water and surface water is exactly azo dyes.In a lot of dyestuffs
The intensive area of industry, river and underground water all suffer from the danger of severe contamination.
Azo dyes also has serious harm to the health of human body.Japanese Yoshida etc. was had found in the thirties in 20th century, molten
Agent Huang can cause mouse canceration of hepatic cell.Hereafter, people just come to realise, during production and use, azo dyes
And its intermediate has danger.Aromatic amine is classified as suspect carcinogen by some countries, wherein as azo dyes intermediate
Beta-naphthylamine and benzidine have been identified as being the carcinogenic substance most strong to the mankind.Wherein, azo group often with one or
Multiple aromatic rings systems are connected, and form conjugated system, so as to the chromogen as dyestuff.He is almost distributed in all colours.
Azo dyes not only can be only used for the printing and dyeing of textile, can also be used to contaminate paper, leather, food etc..It should be noted that
In general, azo dyes itself will not be detrimental to health, but part azo dyes uses aromatic amine intermediate
Synthesis, and aromatic amine intermediate has carcinogenicity, after human body skin long-time is in contact with it, by the new of human normal
Old metabolic process and the material that discharges will be in connection, reduction reaction can occur in conjunction with after, the reaction can cause azo group
Fracture, so as to generate the aromatic amine compounds with carcinogenicity again, if the compound of these generations is inhaled again by human body
Receive, after activation, the cell of human body will change in terms of 26S Proteasome Structure and Function, be transformed into human lesion
Inducement.As a result, causing the possibility of cancer.
Azo dyes gold orange II product water solubles, the mankind in serious harm for pollutions of the azo dyes gold orange II to environment
Health, its pollute environment the comprehensive regulation, there are no the technology of comparative maturity and stability and high efficiency.Gold orange II efficient degrading bacterial strains
Resource is most deficient.
Content of the invention
The purpose of the present invention is：The microbial inoculum of a kind of gold orange II degradation bacterias AO7-2 and its production, degradation bacteria strains AO7-2 are provided
Can efficiently be degraded gold orange II so as to percent of decolourization more than 90%, for related dye wastewater biological intensive treatment and water body or soil
The pollution control of earth, enriches gold orange II degradation bacteria strains resources banks.
The present invention technical solution be：The bacterial strain of gold orange II degradation bacterias AO7-2 is Arthrobacter
Protophormiae, belongs to micrococcaceae, Arthrobacter, is preserved in China typical culture collection within 19th in September in 2016
The heart, address：Wuhan University of Wuhan, China city, deposit number：CCTCC M2016502, Main Biological：Microorganism is in
Spherical, dispersed arrangement, bacterium colony are in faint yellow, circular, moistening, glossy, opaque, surface elevation, neat in edge, and bacterium is without pod
Film, without gemma, Gram's staining is that the positive, bacterium are bred by fragmentation, and the bacterium is heterotrophic bacterium, in growth course
In need oxygen, it is not necessary to sunlight, catalase reacting positive, oxydase reaction are negative, and the optimum growth temperature of the bacterium is 30-
38 C, environmental optima pH are 8.0.
Microbial inoculum is produced using gold orange II degradation bacterias AO7-2, the microbial inoculum is prepared by the following method and forms, it is characterised in that should
Method is comprised the following steps：
Step (1)：During the test tube kind that ring gold orange II degradation bacterias AO7-2 are scraped with oese is inoculated in, 30 DEG C, 180rpm vibrates
Cultivate to logarithmic phase；Wherein, in 1L fermentation mediums, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride
0.5%, remaining deionized water, pH are 9.0；
Step (2)：By step（1）Cultured bacterial classification is inoculated with into 500L seeding tanks by the 1% of culture volume, is cultivated to logarithm
Growth period；In 1L seed tank culture bases, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride 0.5%, remaining
Deionized water, pH are 9.0；Condition of culture is：Lead to filtrated air, the filtrated air being passed through per minute and culture medium in incubation
The ratio of volume is 1:0.6 ~ 1.2, mixing speed is 180 ~ 240r/min, and cultivation temperature is 30 DEG C, and incubation time is 48 ~ 60h；
Step (3)：By step（2）Seed liquor press culture volume 10% access production tank fermented and cultured；The training used of production tank
Foster base is identical with seed tank culture base；Condition of culture is：In incubation lead to filtrated air, the filtrated air being passed through per minute with
The ratio of culture volume is 1:0.6 ~ 1.2, mixing speed be 180 ~ 240r/min, cultivation temperature be 30 DEG C, incubation time be 48 ~
60h；
Step (4)：In production tank, the fermentation ends when thalline quantity reaches 1,000,000,000/more than mL, after the completion of fermentation, nutrient solution goes out
Tank, is directly distributed into liquid agent with plastic barrel or Packaging Bottle, and the liquid agent is microbial inoculum.
The invention has the beneficial effects as follows：1st, degradation bacteria AO7-2 can make the residual quantity of gold orange II reduce by more than 90%, in short-term
In make azo dyes gold orange II be degraded；2nd, using gold orange II degradation bacterias AO7-2 production microbial inoculum, production and application low cost,
Easy to use, removal effect is good, is suitable for the pollution control of related dye wastewater biological intensive treatment and water body and soil, for guarantor
Shield ecological environment, protection human body health, the added value of raising agricultural product have great importance.
Description of the drawings
Fig. 1 is degradation process of strains A O7-2 to gold orange II；
Fig. 2 is impact of the temperature to AO7-2 degradation efficiencies；
Fig. 3-1 is the impact that pH is grown to strains A O7-2；
Fig. 3-2 is impacts of the pH to AO7-2 degradation efficiencies；
Fig. 3-3 is absorption of strains A O7-2 to dyestuff gold orange II under the conditions of serial pH；
Fig. 4 is impact of the salinity to AO7-2 degradation efficiencies；
Fig. 5 is impact of the metal ion to AO7-2 degradation efficiencies.
Specific embodiment
Technical solution of the present invention is further analyzed with reference to embodiment and its accompanying drawing, but be not to be construed as right
The restriction of technical scheme.
Gold orange II degradation bacterias AO7-2, its bacterial strain is Arthrobacter protophormiae, belongs to micrococcaceae, section
Bacillus, is preserved in China typical culture collection center, address on 19th in September in 2016：Wuhan University of Wuhan, China city, protects
Hide numbering：CCTCC M2016502.
1. the separation and identification of bacterial strain：Contaminated soil 10g is taken, is placed in the sterilized water of 100mL, vibrated 5min, obtain soil
Bacteria suspension；The soil bacteria suspension of 5mL is taken, the minimal medium [NH of 100mL is added to4NO31g/L, KH2PO41.5g/L,
K2HPO40.5 g/L, NaCl 0.5g/L, MgSO47H2O, 0.2g/L] in, add gold orange II as carbon source, be placed in 30
DEG C, 150r/min shaking tables shaken cultivation obtains pregnant solution；Gradient dilution pregnant solution coats the culture medium flat plate for adding gold orange II
On, until obtaining the degradation bacteria strains of gold orange II；The degradation bacteria is named as AO7-2, is further purified and is accredited asArthrobacter protophormiae；AO7-2 bacterial strain Main Biologicals：Microorganism be in spherical, dispersed arrangement, bacterium
Fall in faint yellow, circular, moistening, glossy, opaque, surface elevation, neat in edge, bacterium without pod membrane, without gemma, gram
Dye as the positive, bacterium is bred by fragmentation, the bacterium is heterotrophic bacterium, in necessary for growth oxygen, is not required to
Sunlight is wanted, catalase reacting positive, oxydase reaction are negative, the optimum growth temperature of the bacterium is 30-38 C, environmental optima pH
For 8.0；Up to more than 90%, the bacterium can use general of fermentation industry to the degradation rate of the bacterium degraded gold orange II in laboratory conditions
Ferment equipment is produced.
2. laboratory biological degradation experiment
Degradation process of 2.1 strains As O7-2 to gold orange II：Strains A O7-2 is in LB culture mediums（LB culture medium prescriptions：Peptone 1%,
Yeast extract 0.5%, sodium chloride 0.5%,）Middle vibration（30 DEG C, 180rpm）Overnight incubation, it is dilute that next day takes bacterial strain seed liquor liquid feeding body LB
Release 3 times and add gold orange II mother liquors afterwards so as to final concentration of 20mg/L；The degraded system is positioned over quiet in 37 DEG C of constant incubators
Put, and sample at timed intervals（Time interval is 2 hours）, samples taken be placed in mixture of ice and water preserve, treat all samples
It is uniformly processed after taking；After all samples take, through 12000rpm, 3min centrifugations remove thalline, obtain containing being not yet degraded
The supernatant of the gold orange II of decolouring；Supernatant measures its OD value, experiment at 484nm using ultraviolet-uisible spectrophotometer
3 repetitions are set, and mean value is calculated after detection absorbance respectively and the residual quantity of gold orange II in measurement system is come with this；As a result such as
Shown in Fig. 1：Prolongation over time, strains A O7-2 can quickly make gold orange II solution decolourize, and in 24 hours, percent of decolourization exceedes
90%, judge that strains A O7-2 has good degradation and decolorization performance to gold orange II accordingly.
Impact of 2.2 temperature to AO7-2 degradation efficiencies：Temperature has the shadow of highly significant to microbial growth and metabolism
Ring, for illustrating impact of the temperature to strains A O7-2 degraded gold orange II, degraded system will be set up after bacterial strain incubated overnight（With 2.1）,
The degraded system is divided into 6 parts, per part arranges three repetitions；By divide equally after mixed liquor be respectively placed in 10 DEG C, 20 DEG C,
30 DEG C, 37 DEG C, 50 DEG C, in 60 DEG C of constant incubators, sample is taken out in temperature bath after 10 hours, centrifugation is determined respectively and calculates difference
At a temperature of strains A O7-2 azo dyes gold orange II degradation rate（Sample treatment and detection method same 2.1）；As a result such as Fig. 2 institutes
Show：Low temperature and high temperature are respectively provided with significant inhibitory action to strains for degrading gold orange II, and 30 ~ 37 DEG C is the suitable of strains for degrading gold orange II
Suitable temperature range；When temperature meets or exceeds 50 DEG C, bacterial strain is extremely low to the degradation rate of gold orange II, and which can hardly be made to decolourize.
Impacts of 2.3 pH to AO7-2 degradation efficiencies：PH equally has the shadow of highly significant to microbial growth and metabolism
Ring, microorganism all has the pH scopes of a suitable growth and metabolism, surpass and go beyond the scope, microorganism cannot grow well
And metabolism；For investigating the impact that pH grows to strains A O7-2, LB liquid medium is adjusted with hydrochloric acid or sodium hydroxide solution respectively
To pH3 ~ pH10；By 1% inoculum concentration, the LB culture mediums of the AO7-2 seed liquors of incubated overnight in LB to 20mL series pH value are seeded in
In, 30 DEG C are placed in, in 180rpm constant-temperature tables, shaken cultivation was surveyed at 600nm with ultraviolet-uisible spectrophotometer after about 6 hours
Determine bacterium solution light absorption value, and the cell concentration of counter sample is weighed with OD600 values；As a result as shown in figure 3-1：The result of Fig. 3-1
Show bacterial strain well-grown under neutral meta-alkali environment, the advantageous pH range of strain growth is 6 ~ 9；Slant acidity environment and strong basicity
Environment has extremely strong inhibitory action to the growth of bacterial strain, and when pH is less than or equal to 5 or is more than or equal to 10, bacterial strain normally cannot be given birth to
Long.
Overnight culture by strains A O7-2 in LB is divided into 8 parts, 12000rpm, and 3min is collected by centrifugation thalline, and divides
Not Yong equal-volume series pH value the resuspended body of LB nutrient solutions, add the gold orange II mother liquors of final concentration of 20mg/L, the drop for building
Enzymatic hydrolysis system is placed in 37 DEG C of constant incubators, and temperature bath took out sample after 10 hours, is calculated and is analyzed bacterial strain pair under condition of different pH
The degradation rate of gold orange II（Sample treatment and detection method same 2.1）, to study impacts of the pH to strains A O7-2 degraded gold orange II；
As a result as shown in Fig. 3-2 and 3-3；Fig. 3-2 shows that the clearance of gold orange II in mixed liquor in the range of pH3-5 is gradually reduced；pH6-9
When clearance be stepped up, during pH10, clearance slightly reduces；But in slant acidity environment, strains A O7-2 has bright to gold orange II
Aobvious suction-operated, from Fig. 3-3 it can be seen that pH is lower, the concentration of bacterial strain absorbing dye is bigger, and color is also deeper；Comprehensive
In mixed liquor, dyestuff residual and thalline adsorb situation, judge that bacterial strain has preferably degraded effect in slight alkali environment to gold orange II
Really, optimal pH is 9.
Impact of 2.4 salinity to AO7-2 degradation efficiencies：Salinity is frequent in Environmental Quality Evalution and sewage disposal process
One of important parameter of examination, has considerable influence to the growth and metabolism of microorganism；For studying life of the salinity to strains A O7-2
Long impact, by 1% inoculum concentration, is seeded in the AO7-2 seed liquors of incubated overnight in LB respectively to 20mL salinity containing series（0%,
0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%）LB culture mediums in, be placed in 30 DEG C, shake in 180rpm constant-temperature tables
After swinging culture about 6 hours, bacterium solution light absorption value is determined with ultraviolet-uisible spectrophotometer at 600nm, and weighed with OD600 values
The cell concentration of counter sample；Overnight culture by strains A O7-2 in LB is divided into 10 parts, 12000rpm, and 3min is centrifuged
Collects thalline, and the resuspended body of the LB nutrient solutions respectively with equal-volume containing serial salinity, add the gold orange of final concentration of 20mg/L
II mother liquors, are placed in 37 DEG C of constant incubators, and temperature bath took out sample after 10 hours, calculated and bacterium under the different salt concentration conditions of analysis
Degradation rate of the strain to gold orange II（Sample treatment and detection method same 2.1）, to study salinity to strains A O7-2 degraded gold orange II
Impact；As a result as shown in Figure 4：With the raising of salinity, the growth of strains A O7-2 is significantly suppressed, to gold orange II
Degradation rate be also declined slightly, but affect not notable.
Impact of 2.5 metal ions to AO7-2 degradation efficiencies：Metal ion has important shadow to the growth and metabolism of microorganism
Ring, also have considerable influence to the catalysis activity of the various metalloenzyme in microbial body；For studying different metal ions to bacterial strain
The impact of AO7-2 degradation of dye gold orange II, adds a series of different gold in the strain cultured solution containing 20mg/L gold orange II
Category ion（Mg2+、Fe2+、Fe3+、Cu2+、Ca2+、Zn2+、Hg2+、Co2+、Ag1+、Al3+、K1+、Mn2+）, and be respectively provided with 0.1mM,
Tri- concentration gradients of 1mM and 10mM；System is placed in 37 DEG C of constant incubators, and temperature bath took out sample after 10 hours, is calculated and is divided
Analyse relative degradation rate of the different metal ions to strains for degrading gold orange II under each concentration（Sample treatment and detection method same 2.1）；
As a result as shown in figure 5, Mg2+、Ca2+、Al3+、K1+、Mn2+Plasma is had no significant effect to the degradation rate of bacterial strain, and Fe2+、Fe3+、
Co2+Deng there is certain inhibitory action in higher concentration to strains for degrading rate, with its concentration increasing inhibitory action increasingly
By force；Cu2+、Hg2+、Ag1+There is very strong inhibitory action to the degradation rate of bacterial strain；Especially Hg2+, which is at low concentrations just to bacterial strain
Degraded gold orange II has strong inhibitory action.
Production example 1：According to the microbial inoculum that following steps produce gold orange II degradation bacterias AO7-2, the microbial inoculum is by the following method
It is prepared from, it is characterised in that the method is comprised the following steps：
Step (1)：During the test tube kind that ring gold orange II degradation bacterias AO7-2 are scraped with oese is inoculated in, 30 DEG C, 180rpm vibrates
Cultivate to logarithmic phase；Wherein, in 1L fermentation mediums, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride
0.5%, remaining deionized water, pH are 9.0；
Step (2)：By step（1）Cultured bacterial classification is inoculated with into 500L seeding tanks by the 1% of culture volume, is cultivated to logarithm
Growth period；In 1L seed tank culture bases, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride 0.5%, remaining
Deionized water, pH are 9.0；Condition of culture is：Lead to filtrated air, the filtrated air being passed through per minute and culture medium in incubation
The ratio of volume is 1:0.6, mixing speed is 180r/min, and cultivation temperature is 30 DEG C, and incubation time is 48h；
Step (3)：By step（2）Seed liquor press culture volume 10% access production tank fermented and cultured；The training used of production tank
Foster base is identical with seed tank culture base；Condition of culture is：In incubation lead to filtrated air, the filtrated air being passed through per minute with
The ratio of culture volume is 1:0.6, mixing speed is 180r/min, and cultivation temperature is 30 DEG C, and incubation time is 48h；
Step (4)：In production tank, the fermentation ends when thalline quantity reaches 1,000,000,000/more than mL, after the completion of fermentation, nutrient solution goes out
Tank, is directly distributed into liquid agent with plastic barrel or Packaging Bottle, and the liquid agent is microbial inoculum.
Production example 2：According to the microbial inoculum that following steps produce gold orange II degradation bacterias AO7-2, the microbial inoculum is by the following method
It is prepared from, it is characterised in that the method is comprised the following steps：
Step (1)：During the test tube kind that ring gold orange II degradation bacterias AO7-2 are scraped with oese is inoculated in, 30 DEG C, 180rpm vibrates
Cultivate to logarithmic phase；Wherein, in 1L fermentation mediums, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride
0.5%, remaining deionized water, pH are 9.0；
Step (2)：By step（1）Cultured bacterial classification is inoculated with into 500L seeding tanks by the 1% of culture volume, is cultivated to logarithm
Growth period；In 1L seed tank culture bases, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride 0.5%, remaining
Deionized water, pH are 9.0；Condition of culture is：Lead to filtrated air, the filtrated air being passed through per minute and culture medium in incubation
The ratio of volume is 1:0.9, mixing speed is 210r/min, and cultivation temperature is 30 DEG C, and incubation time is 54h；
Step (3)：By step（2）Seed liquor press culture volume 10% access production tank fermented and cultured；The training used of production tank
Foster base is identical with seed tank culture base；Condition of culture is：In incubation lead to filtrated air, the filtrated air being passed through per minute with
The ratio of culture volume is 1:0.9, mixing speed is 210r/min, and cultivation temperature is 30 DEG C, and incubation time is 54h；
Step (4)：In production tank, the fermentation ends when thalline quantity reaches 1,000,000,000/more than mL, after the completion of fermentation, nutrient solution goes out
Tank, is directly distributed into liquid agent with plastic barrel or Packaging Bottle, and the liquid agent is microbial inoculum.
Production example 3：According to the microbial inoculum that following steps produce gold orange II degradation bacterias AO7-2, the microbial inoculum is by the following method
It is prepared from, it is characterised in that the method is comprised the following steps：
Step (1)：During the test tube kind that ring gold orange II degradation bacterias AO7-2 are scraped with oese is inoculated in, 30 DEG C, 180rpm vibrates
Cultivate to logarithmic phase；Wherein, in 1L fermentation mediums, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride
0.5%, remaining deionized water, pH are 9.0；
Step (2)：By step（1）Cultured bacterial classification is inoculated with into 500L seeding tanks by the 1% of culture volume, is cultivated to logarithm
Growth period；In 1L seed tank culture bases, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride 0.5%, remaining
Deionized water, pH are 9.0；Condition of culture is：Lead to filtrated air, the filtrated air being passed through per minute and culture medium in incubation
The ratio of volume is 1:1.2, mixing speed is 240r/min, and cultivation temperature is 30 DEG C, and incubation time is 60h；
Step (3)：By step（2）Seed liquor press culture volume 10% access production tank fermented and cultured；The training used of production tank
Foster base is identical with seed tank culture base；Condition of culture is：In incubation lead to filtrated air, the filtrated air being passed through per minute with
The ratio of culture volume is 1:1.2, mixing speed is 240r/min, and cultivation temperature is 30 DEG C, and incubation time is 60h；
Step (4)：In production tank, the fermentation ends when thalline quantity reaches 1,000,000,000/more than mL, after the completion of fermentation, nutrient solution goes out
Tank, is directly distributed into liquid agent with plastic barrel or Packaging Bottle, and the liquid agent is microbial inoculum.
Above-described embodiment 1-3 gained microbial inoculum degraded gold orange II effect such as following tables：
.
Claims (2)
1. gold orange II degradation bacterias AO7-2, it is characterised in that：The bacterial strain is Arthrobacter protophormiae, belongs to micro-
Coccaceae, Arthrobacter, are preserved in China typical culture collection center, address on 19th in September in 2016：Wuhan, China city is military
Chinese university, deposit number：CCTCC M2016502, Main Biological：In spherical, dispersed arrangement, bacterium colony is in microorganism
Faint yellow, circular, moistening, glossy, opaque, surface elevation, neat in edge, bacterium without pod membrane, without gemma, Gram's staining
For the positive, bacterium is bred by fragmentation, and the bacterium is heterotrophic bacterium, in necessary for growth oxygen, it is not necessary to sun
Light, catalase reacting positive, oxydase reaction are negative, and the optimum growth temperature of the bacterium is 30-38 C, and environmental optima pH is
8.0.
2. with gold orange II degradation bacterias AO7-2 as claimed in claim 1 produce microbial inoculum, the microbial inoculum be prepared by the following method and
Into, it is characterised in that the method is comprised the following steps：
Step (1)：During the test tube kind that ring gold orange II degradation bacterias AO7-2 are scraped with oese is inoculated in, 30 DEG C, 180rpm vibrates
Cultivate to logarithmic phase；Wherein, in 1L fermentation mediums, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride
0.5%, remaining deionized water, pH are 9.0；
Step (2)：By step（1）Cultured bacterial classification is inoculated with into 500L seeding tanks by the 1% of culture volume, is cultivated to logarithm
Growth period；In 1L seed tank culture bases, the weight content of each component is：Peptone 1%, yeast extract 0.5%, sodium chloride 0.5%, remaining
Deionized water, pH are 9.0；Condition of culture is：Lead to filtrated air, the filtrated air being passed through per minute and culture medium in incubation
The ratio of volume is 1:0.6 ~ 1.2, mixing speed is 180 ~ 240r/min, and cultivation temperature is 30 DEG C, and incubation time is 48 ~ 60h；
Step (3)：By step（2）Seed liquor press culture volume 10% access production tank fermented and cultured；The training used of production tank
Foster base is identical with seed tank culture base；Condition of culture is：In incubation lead to filtrated air, the filtrated air being passed through per minute with
The ratio of culture volume is 1:0.6 ~ 1.2, mixing speed be 180 ~ 240r/min, cultivation temperature be 30 DEG C, incubation time be 48 ~
60h；
Step (4)：In production tank, the fermentation ends when thalline quantity reaches 1,000,000,000/more than mL, after the completion of fermentation, nutrient solution goes out
Tank, is directly distributed into liquid agent with plastic barrel or Packaging Bottle, and the liquid agent is microbial inoculum.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115141782A (en) * | 2022-08-16 | 2022-10-04 | 浙江省农业科学院 | Sulfamethazine degrading microbial inoculum and preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101935625A (en) * | 2010-06-12 | 2011-01-05 | 南京农业大学 | Triethylamine degrading bacterium and microbial agent produced therefrom |
| CN103103167A (en) * | 2013-01-29 | 2013-05-15 | 河北师范大学 | Mutant zymoprotein of D-amino acid oxidase and preparation method of mutant zymoprotein |
| CN104195065A (en) * | 2014-07-03 | 2014-12-10 | 杭州电子科技大学 | Orange II degrading bacterium L-15 and inocula produced from same |
| CN104195066A (en) * | 2014-07-03 | 2014-12-10 | 杭州电子科技大学 | Azo dye orange II degrading bacterium and inocula produced from same |
| CN104328068A (en) * | 2014-09-30 | 2015-02-04 | 中南民族大学 | Arthrobacter protophormiae strain, screening method and uses thereof |
-
2016
- 2016-11-03 CN CN201610952903.9A patent/CN106497837B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101935625A (en) * | 2010-06-12 | 2011-01-05 | 南京农业大学 | Triethylamine degrading bacterium and microbial agent produced therefrom |
| CN103103167A (en) * | 2013-01-29 | 2013-05-15 | 河北师范大学 | Mutant zymoprotein of D-amino acid oxidase and preparation method of mutant zymoprotein |
| CN104195065A (en) * | 2014-07-03 | 2014-12-10 | 杭州电子科技大学 | Orange II degrading bacterium L-15 and inocula produced from same |
| CN104195066A (en) * | 2014-07-03 | 2014-12-10 | 杭州电子科技大学 | Azo dye orange II degrading bacterium and inocula produced from same |
| CN104328068A (en) * | 2014-09-30 | 2015-02-04 | 中南民族大学 | Arthrobacter protophormiae strain, screening method and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| ASHVINI CHAUHAN 等: "Degradation of o-Nitrobenzoate via Anthranilic Acid (o-Aminobenzoate) by Arthrobacter protophormiae:A Plasmid-Encoded New Pathway", 《BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115141782A (en) * | 2022-08-16 | 2022-10-04 | 浙江省农业科学院 | Sulfamethazine degrading microbial inoculum and preparation method and application thereof |
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