CN105087440B - Pseudomonas mendocina NX-1 and its application in n-hexane degradation - Google Patents
Pseudomonas mendocina NX-1 and its application in n-hexane degradation Download PDFInfo
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Abstract
The present invention provides one plant of new high-efficiency n-hexane degradation bacterias --- pseudomonas mendocina (Pseudomonas mendocina) NX 1 and its application in microbial degradation n-hexane；Pseudomonas mendocina NX 1 is preserved in China typical culture collection center, address：Wuhan, China university, 430072, preservation date：On March 15th, 2015, deposit number：CCTCC NO：M2015114；N-hexane degradation bacteria provided by the invention is aerobic non-fermented type Gram-negative bacterium, can by the use of n-hexane as sole carbon source breeding and by the substrate permineralization into CO2And H2O；Under the conditions of pure culture, can degrade n-hexane in the range of the bacterium pH 4.0~10.0；The bacterial strain has stronger adaptive capacity to environment, and the engineer application for biological method purification waste water containing n-hexane and exhaust gas is laid a good foundation.
Description
(1) technical field
The present invention relates to one plant of new high-efficiency n-hexane degradation bacterias --- pseudomonas mendocina (Pseudomonas
Mendocina) NX-1 and its application in microbial degradation n-hexane.
(2) background technology
N-hexane (n-hexane, C6H6) it is a kind of colourless liquid that is less toxic, having faint special odor.It is a kind ofization
Learn solvent, be mainly used for solvent of the propylene when olefinic polymerizations, the extractant of edible vegetable oil, the solvent of rubber and coating and
The diluent of pigment has certain toxicity, human body can be entered by approach such as respiratory tract, skins, Long Term Contact can cause people
There is headache, the slow poisonings symptom such as dizzy, weak, numb limb in body, serious to cause to fall in a swoon, is unconsciousness, even extremely
It dies.
Extensive use and long-term sucking n-hexane due to n-hexane can cause slow poisoning, develop economical and have reality
The method of the processing n-hexane of effect has become the research hotspot of field of environment pollution control.The biodegradation of n-hexane is demonstrate,proved
It is bright be it is feasible, such as using bio-trickling filter handle high load capacity n-hexane exhaust gas, reached the degradation rate of 70%-90%.Although
In this way, it is domestic at present less to the biodegradation research of n-hexane, it seldom has been reported that and filters out using n-hexane as sole carbon source
Efficient degrading bacterial strain.The present invention screens the bacterial strain of one plant of energy efficient degradation n-hexane from environment, for containing the pollutant
The biological cleaning engineer application of waste water and exhaust gas lays the foundation.
Microorganism is that a kind of species is more, and breeding is fast, adaptable, the strong organism of metabolic capability.If it can screen and can divide
The microorganism of efficient degradation n-hexane is separated out, n-hexane is degraded into the substance to human body and environment nonhazardous, this is to maintaining people
Anti- and ecological safety is had profound significance by class.
(3) content of the invention
It is an object of the invention to provide one plant of new and effective n-hexane degradation bacterias --- pseudomonas mendocina NX-1 and
Its application in microbial degradation n-hexane.
The technical solution adopted by the present invention is：
Pseudomonas mendocina (Pseudomonas mendocina) NX-1, is preserved in China typical culture collection
The heart, preservation date be on March 15th, 2015, deposit number：CCTCC NO:M2015114, address：China, Wuhan, Wuhan are big
It learns, postcode 430072.
The Genebank accession number of the 16s rDNA of the pseudomonas mendocina NX-1 be KP938962, the master of the bacterial strain
The biological property is wanted to be：Thalline be in ellipticity, size be (0.3~0.6) μ m (0.8~2.0) μm, no gemma, amphitrichous；Bacterium
It is in ellipticity to fall, transparent, form is full, smooth moistening, is easily provoked, and lawn is grown along line；Aerobic, oxydase reaction is sun
Property, indole reaction, contact enzyme reaction, citrate are the positive；M.R reactions, V.P. reactions are feminine gender；Sugared fermenting experiment is negative,
Gram-negative.
The invention further relates to applications of the pseudomonas mendocina NX-1 in microbial degradation n-hexane, specifically
, the application is：By pseudomonas mendocina NX-1 through seed culture medium (preferably containing 100~400mg/L of final concentration just oneself
The seed culture medium of alkane) culture gained bacterium solution as enzyme source, using n-hexane as substrate, in minimal medium, in pH=4
~10,20 DEG C~40 DEG C, under the conditions of 100~200rpm (preferably pH=6~9,25 DEG C~35 DEG C, 100~200rpm) dropped
Solution reaction, realizes the degradation to n-hexane, and preferred seed culture medium is minimal medium.
The substrate n-hexane in minimal medium initial concentration for 20~800mg/L minimal mediums (preferably
100~400mg/L), enzyme source dosage is with OD in minimal medium600It is calculated as 0.01~0.03.
The minimal medium final concentration composition is as follows：Na2HPO4 4.5g/L、KH2PO4 1.0g/L、(NH4)2SO42.5g/L、MgSO4·7H2O 0.2g/L、CaCl20.023g/L, micro- mother liquor 1mL/L, pH 7.0, solvent are to go
Ionized water；The micro- mother liquid concentration composition：FeSO4·7H2O 1.0g/L、CuSO4·5H2O 0.02g/L、H3BO3
0.014g/L、MnSO4·4H2O 0.10g/L、ZnSO4·7H2O 0.10g/L、Na2MoO4·2H2O 0.02g/L、CoCl2·
6H2O 0.02g/L, solvent are deionized water.
Enzyme source of the present invention prepares as follows：
1) inclined-plane culture：Pseudomonas mendocina NX-1 activation is inoculated in R2A solid slope culture mediums, 30~32 DEG C of trainings
36~48h is supported, obtains inclined-plane thalline, the R2A solid slope culture mediums final concentration forms：Dusty yeast 0.50g/L, casein
0.50g/L, soluble starch 0.50g/L, MgCl2·7H2O 0.05g/L, tryptone 0.50g/L, glucose 0.50g/L, third
Ketone acid sodium 0.30g/L, KH2PO40.45g/L, agar 15~18g/L, pH 7.2, solvent is water；
2) seed culture：The inclined-plane thalline obtained with oese picking step (1) is seeded to containing 100~400mg/ of final concentration
The seed culture medium culture of L n-hexanes 30~32 DEG C, cultivates 36~38h, obtains bacterium solution；The seed culture medium is inorganic salts
Culture medium.
The beneficial effects are mainly as follows：The present invention provides the efficient degrading bacterias of one plant of n-hexane -- Mendoza
Pseudomonad NX-1, the bacterial strain are aerobic non-fermented type Gram-negative bacterium, can be using n-hexane as sole carbon source and energy
The source growth efficient degradation substrate simultaneously, n-hexane concentration that can be within degradable up to 700mg/L, and the bacterial strain
Optimum growth temperature and pH scopes are wide, and at 30~37 DEG C, pH can preferably degrade n-hexane for 6.0~9.0, and degradation rate can
Reach more than 80%.Based on bacterial strain NX-1 for the efficient degradation rate of n-hexane so that small investment, operating cost are low, do not generate
Secondary pollution, the method for easy to operate, stable Biochemical method high load capacity n-hexane exhaust gas are than costly physics
The degradation effect of method and chemical method is even better.The present invention establishes for the engineer application of biological method purification waste water containing n-hexane and exhaust gas
Basis is determined.
(4) illustrate
Fig. 1 is the transmission electron microscope photo of pseudomonas mendocina NX-1；
Fig. 2 is phylogenetic tree；
Fig. 3 is the thalli growth of pseudomonas mendocina NX-1, n-hexane degradation change curve；
Fig. 4 is influence column diagram of the different pH value culture solutions to the n-hexane degradation rate of pseudomonas mendocina NX-1；
Fig. 5 is influence column diagram of the different temperatures to the n-hexane degradation rate of pseudomonas mendocina NX-1；
Fig. 6 is degradation curve figures of the pseudomonas mendocina NX-1 to different initial concentration n-hexanes；
Fig. 7 is growth curves of the pseudomonas mendocina NX-1 under different initial n-hexane concentration；
Fig. 8 purifies n-hexane waste water efficiency curve diagram for pseudomonas mendocina NX-1；
Fig. 9 purifies n-hexane exhaust gas efficiency curve diagram for pseudomonas mendocina NX-1.
(5) specific embodiment
With reference to specific example, the present invention is described further, but protection scope of the present invention is not limited in
This：
The minimal medium concentration forms：Na2HPO4·12H2O 4.5g/L、KH2PO4 1.0g/L、(NH4)2SO4
2.5g/L、MgSO4·7H2O 0.2g/L、CaCl20.023g/L, micro- mother liquor 1mL/L, pH 7.0, solvent is water, institute
State micro- mother liquor composition：FeSO4·7H2O 1.0g/L、CuSO4·5H2O 0.02g/L、H3BO3 0.014g/L、
MnSO4·4H2O 0.10g/L、ZnSO4·7H2O 0.10g/L、Na2MoO4·2H2O 0.02g/L、CoCl2·6H2O 0.02g/
L, solvent are deionized water.
The R2A solid mediums final concentration forms：Dusty yeast 0.50g/L, casein 0.50g/L, soluble starch
0.50g/L、MgCl2·7H2O 0.05g/L, tryptone 0.50g/L, glucose 0.50g/L, Sodium Pyruvate 0.30g/L,
KH2PO40.45g/L, agar 15~18g/L, pH 7.2, solvent is water.
Embodiment 1：The separation and identification of pseudomonas mendocina NX-1
(1) sample collection and domestication
Primary dcreening operation：The activated sludge of collection in worksite Zhejiang pharmaceutical chemical industry factory treatment tank after standing sedimentation 2h, removes
Supernatant liquor and suspended impurity leave the tiny sludge of particle, take lower floor's sludge after standing with minimal medium by 1:1
(v/v) mixing accesses to sludge acclimatization tank, total volume 5L, is substrate as sole carbon source and the energy using n-hexane, and (30 ± 1
DEG C) constant temperature domestication culture, sludge is tamed, it is stable aerated to domestication tank progress during domestication experiment, control domestication tank
PH value maintains 7.0, daily when concentration of substrate drops to below 50mg/L, adds a substrate so that concentration of substrate reaches after adding in
To 150mg/L (sampling detection residual substrate concentration before substrate adds), interval 3d replaces mixed culture medium, nearly after two months,
The sludge of domestication can stablize degradation 150mg/L n-hexanes daily, this sample is transferred further to be enriched with into shaking flask, obtain domestication sample
Product.
(2) will domestication sample be seeded to 50mL minimal mediums by the access amount of volumetric concentration 10%, using n-hexane as
Substrate, initial concentration 105mg/L, treat that substrate drops by 30 DEG C, cultivate on 160r/min shaking tables and detect residual substrate concentration afterwards for 24 hours
Solution rate reaches 80% or so, and the fresh inorganic salts that the n-hexane containing equal concentrations is transferred to the access amount of volumetric concentration 10% are trained
It supports in base, after carrying out repeatedly passage enrichment, is diluted coating, picking single bacterium colony, then using n-hexane as substrate, carries out degradation work
Property measure, isolate and purify, obtain one plant have n-hexane degrading activity bacterial strain NX-1.
(2) isolation and identification for strains
Secondary screening and purifying：The mixed bacteria liquid of repeatedly passage enrichment will be passed through in minimal medium, is diluted coating,
According to the otherness of phage populations, picking single bacterium colony.After repeatedly line separation is carried out to single bacterium colony, then it is connected to using n-hexane as only
In the minimal medium of one carbon source and the energy, degrading activity is tested.Single bacterium colony of the selection with n-hexane degradation capability, into one
Step isolates and purifies, and obtains the pure bacterial strain NX-1 for having n-hexane degrading activity.
The morphological feature of bacterial strain NX-1：Cell thalline is in ellipticity, and size is (0.3~0.6) μ m (0.8~2.0) μm,
Without gemma, amphitrichous；Bacterium colony is in small round shape, transparent, form is full, unstressed configuration, smooth moistening, easily provokes, and lawn is given birth to along line
It is long.
The physiological and biochemical property of bacterial strain NX-1 is：Aerobic, oxydase reaction is the positive, and indole reaction contacts enzyme reaction, lemon
Lemon hydrochlorate is the positive；M.R reactions, V.P. reactions are feminine gender；Sugared fermenting experiment is feminine gender, and Gram's staining is feminine gender.
Examining order is completed by Sangon Biotech (Shanghai) Co., Ltd..The 16S rDNA sequences of bacterial strain are as follows
(Genebank accession number is KP938962), SEQ ID NO.1：
CCAAATGGCGCCGGACTACCATGCAGTCGAGCGGATGAGAGGGGCTTGCTCCTTGATTTAGCGGCGGACGGGTGAGT
AATGCCTAGGAATCTGCCTGGTAGTGGGGGATAACGTTCCGAAAGGAACGCTAATACCGCATACGTCCTACGGGAGA
AAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAATGGCTCACC
AAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGA
GGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGAT
TGTAAAGCACTTTAAGTTGGGAGGAAGGGCATTAACCTAATACGTTAGTGTTTTGACGTTACCGACAGAATAAGCAC
CGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGC
GTAGGTGGTTCGTTAAGTTGGATGTGAAAGCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGGCGAGCTAGAGT
ACGGTAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGA
CCACCTGGACTGATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCC
GTAAACGATGTCAACTAGCCGTTGGGTTCCTTGAGAACTTAGTGGCGCAGCTAACGCATTAAGTTGACCGCCTGGGG
AGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAA
GCAACGCGAAGAACCTTACCTGGCCTTGACATGCTGAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACTCAGA
CACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTC
CTTAGTTACCAGCACGTTATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTC
AAGTCATCATGGCCCTTACGGCCAGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGT
GGAGCTAATCCCATAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTA
ATCGTGAATCAGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTG
CTCCAGAAGTAGCTAGTCTAACCTTCGGGCGACGGTACCACGGAAGGATCCACGA。
Research and analysis are carried out by the morphological feature to bacterial strain NX-1 and physiological and biochemical property, by features described above with
《Bergry’s Manual of Systematic Bacteriology》The Pseudomonas edited and recorded is compared, with reference to bacterial strain NX-1
16S rRNA homology analysis, phylogenetic tree such as Fig. 2, so that it is determined that bacterial strain NX-1 be Pseudomonas
Mendocina is named as pseudomonas mendocina (Pseudomonas mendocina) NX-1.
It is prepared by the seed liquor of 2 pseudomonas mendocina NX-1 bacterial strains of embodiment
1) inclined-plane culture：Pseudomonas mendocina NX-1 is inoculated in R2A solid slope culture mediums, 30~32 DEG C of cultures 36
~48h obtains inclined-plane thalline, the R2A solid slope culture mediums final concentration forms：Dusty yeast 0.50g/L, casein
0.50g/L, soluble starch 0.50g/L, MgCl2·7H2O 0.05g/L, tryptone 0.50g/L, glucose 0.50g/L, third
Ketone acid sodium 0.30g/L, KH2PO40.45g/L, agar 15~18g/L, pH 7.2, solvent is water.
2) seed culture：The inclined-plane thalline obtained with oese picking step (1) is seeded to seed culture medium culture, adds in
The n-hexane of 100~400mg/L of final concentration 30~32 DEG C, cultivates 36~38h, obtains OD600=0.1~0.3 bacterium solution, it is described
Seed culture medium is minimal medium.
Embodiment 3：Pseudomonas mendocina NX-1 (CCTCC NO：M 2015114) degradation n-hexane performance
Using n-hexane as the sole carbon source of pseudomonas mendocina NX-1, the bacterium solution of 2 method of Example preparation, inoculation
To the 50mL minimal mediums of the fresh n-hexane containing 105mg/L, make initial cell concentration (with OD600Meter) it is 0.02, pH=
7.It is put into the shaking table that temperature is 30 DEG C, revolution is 160r/min and cultivates, n-hexane degradation rate is measured by sampling every 3h~4h, and
A part of bacterium solution is extracted out with 5mL syringes, measures thalline OD values, the result is shown in Fig. 3.In experimentation, 2 Duplicate Samples and one are designed
A blank control group for not connecing bacterial strain.With the extension of time, cell concentration gradually increases, until during 30h, cell concentration reaches most
About 70mg/L (is converted) with OD.This example demonstrates that pseudomonas mendocina NX-1 can be by the use of n-hexane as sole carbon source
Growth and breeding is carried out with the energy, and with the ability of stability and high efficiency degradation n-hexane.
Influence of the embodiment 4pH values to pseudomonas mendocina NX-1 degradation n-hexanes
With 1mol/L NaOH aqueous solutions or 1mol/L H2SO4Aqueous solution adjust minimal medium for different pH value (4.0,
5.0th, 6.0,6.5,7.0,7.5,8.0,9.0,10.0), under conditions of initial n-hexane concentration is 105mg/L, access is implemented
Bacterium solution prepared by 2 method of example, it is 0.02 to make the initial cell concentration (in terms of OD) in each Duplicate Samples.By sample in 30 DEG C, 160r/
Shaken cultivation in min constant-temperature tables samples after cultivating 30h, n-hexane degradation rate in survey reaction solution, in experimentation, designs 2
Duplicate Samples and a blank control group for not connecing bacterial strain, the result is shown in Fig. 4.As it can be seen that in pH4.0~10.0, microorganism can drop
Solve n-hexane；As pH from 4.0 increases to 10.0, the degradation rate first increases and then decreases of n-hexane, pseudomonas mendocina NX-1
The convenient pH value of n-hexane of degrading is 6.0~9.0.Show that pseudomonas mendocina NX-1 can different journeys in pH6.0~9.0
Degree ground degradation n-hexane, guarantee is provided for its application in different pH environment.
Influence of 5 temperature of embodiment to pseudomonas mendocina NX-1 degradation n-hexanes
In the minimal medium for being 105mg/L in initial n-hexane concentration, the bacterium solution of access embodiment 2 method preparation,
It is 0.02 (in terms of OD) to make the initial cell concentration in each Duplicate Samples.By each sample be respectively placed in temperature for 20 DEG C, 25 DEG C, 30
DEG C, 35 DEG C, 37 DEG C, constant-temperature shaking culture (shaking table revolution is 160r/min) in 40 DEG C of shaking tables, sample, survey anti-after cultivating 30h
It answers n-hexane degradation rate in liquid, in experimentation, designs 2 Duplicate Samples and a blank control group for not connecing bacterial strain.It can by Fig. 5
Know, in 20 DEG C~40 DEG C temperature ranges, as temperature increases to 40 DEG C from 20 DEG C, the degradation of n-hexane takes the lead in subtracting after increase
Small, the pseudomonas mendocina NX-1 convenient temperature of degradation n-hexane is 30 DEG C or so, with temperature further rise or
Decline, the degradation capability of bacterial strain is begun to decline.
Influence of 6 concentration of substrate of embodiment to pseudomonas mendocina NX-1 degradation n-hexanes
Under convenient condition of culture (pH7.0,30 DEG C of temperature), pseudomonas mendocina NX-1 is to various concentration for research
The degradation of n-hexane.The substrate n-hexane of various concentration is added in fresh minimal medium, distinguishes initial substrate concentration
For 50,100,150,200,250,300,400,500,600,700,800mg/L, be inoculated with bacterium prepared by 2 method of embodiment respectively
Liquid, it is 0.02 (in terms of OD) to make the initial cell concentration in each Duplicate Samples.It is trained in 30 DEG C, the shaking table that revolution is 160r/min
It supports, the growth OD and n-hexane degradation rate of timing sampling measure pseudomonas mendocina NX-1, in experimentation, designs 2
Duplicate Samples and a blank control group for not connecing bacterial strain.As a result as knowable to Fig. 6 and Fig. 7, pseudomonas mendocina NX-1 degrades just
The degradation rate of hexane is influenced smaller by initial substrate concentration, it is seen that and n-hexane is smaller to the toxicity of bacterial strain, and in higher concentration
Under still can be by its degradable mineralising, but after the concentration of n-hexane is 800mg/L, just cannot degrade again complete.
Embodiment 7：Pseudomonas mendocina NX-1 purifies n-hexane waste water
Be inoculated with pseudomonas mendocina NX-1 bacterium solutions prepared by embodiment 2 method to certain pharmaceutical factory containing 100mg/L just oneself
The waste water of alkane makes initial cell concentration (with OD600Meter) it is 0.02, pH=7；Concrete operation method is with embodiment 3, by 30h's
After processing, the purification efficiency of n-hexane is up to 100%.The results are shown in Figure 8, by pseudomonas mendocina NX-1 for purify just oneself
Alkane waste water has good application effect.
Embodiment 8：Pseudomonas mendocina NX-1 purifies n-hexane exhaust gas
Be inoculated with the bacterium solution of pseudomonas mendocina NX-1 prepared by embodiment 2 method to bio-trickling filter (referring to Zhang Dingfeng,
Research [J] environmental sciences of the bio-trickling filters such as Fang Junyi, leaf outstanding person's rising sun purification multicomponent exhaust gas, 2013.) in, for purifying
Exhaust gas containing n-hexane, it is 200mg/m to set n-hexane inlet gas concentration3, after the start-up of 20 days, it is in the residence time
Under conditions of 36s, n-hexane removal rate can stable operation always up to more than 92%, hereafter system.The results are shown in Figure 9, Men Duo
The n-hexane exhaust gas performance of Sa pseudomonad NX-1 continuous processing high concentrations in bio-trickling filter is stablized, and purification efficiency is high.
Claims (6)
- Pseudomonas mendocina 1. (Pseudomonas mendocina) NX-1, is preserved in China typical culture collection center, Preservation date be on March 15th, 2015, deposit number：CCTCC NO:M2015114, address：China, Wuhan, Wuhan University, postal Compile 430072.
- 2. a kind of applications of pseudomonas mendocina NX-1 described in claim 1 in microbial degradation n-hexane.
- 3. application as claimed in claim 2, it is characterised in that the application is：By pseudomonas mendocina NX-1 through seed The bacterium solution that medium culture is obtained is as enzyme source, using n-hexane as substrate, in minimal medium, in pH=4~10,20 DEG C~40 DEG C, carry out degradation reaction under the conditions of 100~200rpm, realize the degradation to n-hexane；The minimal medium is whole Concentration composition is as follows：Na2HPO4 4.5g/L、KH2PO4 1.0g/L、(NH4)2SO4 2.5g/L、MgSO4·7H2O 0.2g/L、 CaCl20.023g/L, micro- mother liquor 1mL/L, pH 7.0, solvent is deionized water；The micro- mother liquid concentration Composition：FeSO4·7H2O 1.0g/L、CuSO4·5H2O 0.02g/L、H3BO3 0.014g/L、MnSO4·4H2O 0.10g/L、 ZnSO4·7H2O 0.10g/L、Na2MoO4·2H2O 0.02g/L、CoCl2·6H2O 0.02g/L, solvent are deionized water.
- 4. application as claimed in claim 3, it is characterised in that n-hexane initial concentration in minimal medium is 20 ~800mg/L.
- 5. application as claimed in claim 3, it is characterised in that enzyme source dosage is with OD in minimal medium600It is calculated as 0.01~0.03.
- 6. application as claimed in claim 3, it is characterised in that the enzyme source prepares as follows：1) inclined-plane culture：Pseudomonas mendocina NX-1 is inoculated in R2A solid slope culture mediums, 30~32 DEG C of cultures 36~ 48h obtains inclined-plane thalline, the R2A solid slope culture mediums final concentration forms：Dusty yeast 0.50g/L, casein 0.50g/ L, soluble starch 0.50g/L, MgCl2·7H2O 0.05g/L, tryptone 0.50g/L, glucose 0.50g/L, Sodium Pyruvate 0.30g/L、KH2PO40.45g/L, 15~18g/L of agar；PH is 7.2, and solvent is water；2) seed culture：The inclined-plane thalline obtained with oese picking step (1) is seeded to containing 100~400mg/L of final concentration just In the seed culture medium of hexane, 30~32 DEG C, 36~38h is cultivated, obtains OD600=0.1~0.3 bacterium solution；The seed culture Base is minimal medium.
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| CN111073835B (en) * | 2019-12-28 | 2022-07-12 | 河南省农业科学院植物保护研究所 | Pseudomonas mendocina capable of effectively degrading atrazine and application thereof |
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| CN102533595A (en) * | 2011-12-17 | 2012-07-04 | 浙江工业大学 | Starkeya sp. T-2 and application thereof |
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2015
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101787353A (en) * | 2009-12-24 | 2010-07-28 | 中国水产科学研究院长江水产研究所 | Pseudomonas mendocina CY004 for efficiently removing nitrite nitrogen, nitrate nitrogen and ammonia nitrogen in water body and application thereof |
| CN102168052A (en) * | 2011-01-28 | 2011-08-31 | 黑龙江大学 | Pseudomonas mendocina strain capable of effectively degrading fomesafen |
| CN102533595A (en) * | 2011-12-17 | 2012-07-04 | 浙江工业大学 | Starkeya sp. T-2 and application thereof |
Non-Patent Citations (1)
| Title |
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| 门多萨假单胞菌DS04-T对PHBV降解性能研究;高佳 等;《辽宁石油化工大学学报》;20130930;第33卷(第3期);4-7 * |
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