CN102965309B - Rhodococcus sp. and application thereof to micro-biologically degrading 4-fluorocinnamic acid - Google Patents
Rhodococcus sp. and application thereof to micro-biologically degrading 4-fluorocinnamic acid Download PDFInfo
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- CN102965309B CN102965309B CN201210460643.5A CN201210460643A CN102965309B CN 102965309 B CN102965309 B CN 102965309B CN 201210460643 A CN201210460643 A CN 201210460643A CN 102965309 B CN102965309 B CN 102965309B
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- VFQOFJQVKVEXIY-UHFFFAOYSA-N 3-(phenylmethoxycarbonylamino)-3-piperidin-3-ylpropanoic acid Chemical compound C1CCNCC1C(CC(=O)O)NC(=O)OCC1=CC=CC=C1 VFQOFJQVKVEXIY-UHFFFAOYSA-N 0.000 title claims abstract description 66
- 241000187562 Rhodococcus sp. Species 0.000 title claims abstract description 7
- 230000000593 degrading effect Effects 0.000 title abstract description 6
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Images
Abstract
The invention provides a novel efficient 4-fluorocinnamic acid degrading bacterium-Rhodococcus sp. HZF1 and an application thereof to micro-biologically degrading 4-fluorocinnamic acid. The strain was collected in the China Center for Type Culture Collection (CCTCC), with the collection date being October 15, 2012 and collection number being CCTCC No: M2012404. The address of the CCTCC is Wuhan University, Wuhan, China and the postcode is 430072. The Rhodococcus sp. HZF1 can be applied to degrading 4-fluorocinnamic acid in water and soil by way of direct addition and can safely, efficiently and quickly degrade residual 4-fluorocinnamic acid on such objects as water and soil. The fungicides containing the strain are simple in preparation processes, low in costs and convenient to use and have good application prospects.
Description
(1) technical field
The present invention relates to the new and effective 4-fluoro cinnamic acid of strain degradation bacteria---rhodococcus (Rhodococcus sp.) HZF1, and application in microbiological deterioration 4-fluoro cinnamic acid.
(2) background technology
4-fluoro cinnamic acid (4-Fluorocinnamic acid), also claims fluoro cinnamic acid, is a kind of aromatic fluorine compound, is mainly prepared by fluorine phenylacrolein, propanedioic acid and pyridine, is widely used at present the field such as medicine intermediate, sensitizer.The molecular formula of 4-fluoro cinnamic acid is C
9h
7fO
2, structural formula is as shown in formula I.
Fluorinated organic compound is widely used in industry, agricultural, and is representing the very important environmental pollutant of a class.Fluorinated organic compound is because its dosage is few, toxicity is little, performance good and for agricultural, because its biologically stable, high biological activity, lipophilicity and for the production of medicine.Recently, fluorinated organic compound is widely used in electronic industry, for example polymkeric substance of 4-fluorophenol and 4-fluoro cinnamic acid polymkeric substance, and their stability is high, viscosity is low, in heat, light, strong current, chemical, can keep high voltage.Fluoro organic matter receives less concern than chloro and bromo organism and is generally considered to biologically inert because of them, and people think that they are less on the impact of human health and environment by mistake.But inert molecule has more persistence and Geng Yi accumulation, the environment being polluted by them is just more difficult reparation also.Research shows, some fluoro organic matters can carry out limited bio-transformation under applicable envrionment conditions, and, fluorinated organic compound shows some significant biological effect, as inhibitory enzyme activity, material transfer between block cell, film transport and affect production capacity process etc.Therefore, the ecological migration and conversion of fluorinated organic compound receives publicity just day by day.
A large amount of uses and the biologically inert of 4-fluoro cinnamic acid, make its constantly accumulation in environment, HUMAN HEALTH and environmental safety are caused to very big harm, and main manifestations is inhibitory enzyme activity, affect transmission ofenergy, destroy between cytolemma material transfer etc. brings out the diseases such as cancer, cardiopulmonary infringement, hepatomegaly.
Processing to the sewage containing 4-fluoro cinnamic acid is very important to protection of the environment quality.In all treatment processs, utilize microbial metabolism method to remove 4-fluoro cinnamic acid, cost is low, and harmful byproduct is few, and it can, pollutent permineralization, therefore have broad application prospects.
Microorganism is that a class kind is many, breeding is fast, strong adaptability, organism that metabolic capacity is strong.If energy screening and separating goes out the microorganism of energy efficient degradation 4-fluoro cinnamic acid, 4-fluoro cinnamic acid is degraded into the material to human body and environment toxicological harmless such as carbonic acid gas, water, this has profound significance to safeguarding human health.
(3) summary of the invention
The object of the invention is to provide the new and effective Rhod 4-of strain fluoro cinnamic acid degradation bacteria HZF1 and application thereof.
The technical solution used in the present invention is:
Rhodococcus (Rhodococcus sp.) HZF1, is preserved in Chinese Typical Representative culture collection center, address: China, and Wuhan, Wuhan University, postcode 430072, preservation date is on October 15th, 2012, deposit number is CCTCC No:M 2012404.
The Genbank number of logging in of the 16S rDNA of described rhodococcus HZF1 is JX878615.
Rhodococcus HZF1(Rhodococcus sp. HZF1) screening and the qualification of bacterial strain:
1) substratum
Minimal medium final concentration consists of: in every liter of nutrient solution, contain NaCl 1g, K
2hPO
41.5g, KH
2pO
40.5g, (NH4)
2sO
41.5g, MgSO
40.1g, 1ml trace element solution, solvent is water, natural pH value makes after high pressure steam sterilization (121 DEG C, 20min), wherein in every liter of trace element solution, contains MnSO
4﹒ H
2o 0.13
g, ZnCl
20.23g, CuSO
4﹒ H
2o 0.03g, CoCl
2﹒ 6H
2o 0.42g, Na
2moO
4﹒ 2H
2o 0.15g, AlCl
3﹒ 6H
2o 0.05g, solvent is water.
Enrichment culture liquid: add 4-fluoro cinnamic acid in minimal medium, the final concentration that makes 4-fluoro cinnamic acid is 500mg/L.
LB liquid nutrient medium: in every liter of nutrient solution, contain yeast powder 10g, peptone 5.0g, sodium-chlor 10.0g, solvent is water, natural pH value makes after high pressure steam sterilization (121 DEG C, 20min).
LB solid medium: in every liter of cultivation, contain yeast powder 10g, peptone 5.0g, sodium-chlor 10.0g, agar 15.0g, solvent is water, natural pH value makes after high pressure steam sterilization (121 DEG C, 20min).
2) strains separation purifying
Mud sample picks up from Hangzhou insecticide factory, get 5ml mud sample and be placed in 250ml Erlenmeyer flask, add 100ml enrichment culture liquid, dark shaking culture (30 DEG C, 150rpm) 1 week, gets the turbid liquid in 5ml upper strata in fresh enrichment culture liquid 100 ml, continue (30 DEG C of dark shaking culture, 150rpm) 1 week, repeat aforesaid operations process 3 times, each inoculum of cultivating is all taken from the nutrient solution of cultivating gained last time.
Nutrient solution 5 ml that get last cultivation gained carry out gradient dilution (10
-4, 10
-5, 10
-6), the nutrient solution 150 μ l that get after each dilution coat on the LB solid medium flat board containing 500mg/L4-fluoro cinnamic acid, being placed in constant incubator (30 DEG C) cultivates, until growing on flat board after bacterium colony, the each bacterium colony of picking is in containing purifying repeatedly on the LB solid medium flat board of 500mg/L4-fluoro cinnamic acid, until bacterium colony is single, each bacterium colony after purifying is connected to respectively in LB liquid nutrient medium test tube to (30 DEG C of shaking culture, 150rpm) spend the night, by the centrifugal (8000rpm of cultured bacterium liquid, 5min), be connected to 25 ~ 45 DEG C of cultivation 3d in enrichment culture liquid, detect the residual quantity of 4-fluoro cinnamic acid in each enrichment culture liquid by high performance liquid chromatography (HPLC), finally screening obtains the bacterial strain of a strain energy efficient degradation 4-fluoro cinnamic acid, called after HZF1.
3) identification of strains
The bacterial strain of above-mentioned acquisition is carried out to morphological specificity and molecular biology identification, and the electromicroscopic photograph of this bacterial strain as shown in Figure 1.The main biological property of this bacterial strain is: gram positive, and aerobic-type, without gemma, size is about that (m), bacterium colony is smooth for 1.7 ~ 3.2 μ, intermediate projections, edge-diffusion, be light orange, catalase feminine gender, oxidase negative, the methyl red test positive, can utilize beta-cyclodextrin, starch, glucose, polysorbate40, acetate, voges-Proskauer test feminine gender, V.P. reacting positive.The optimum growth conditions of this bacterial strain is pH value 7.0,30 DEG C of temperature.This bacterial strain is accredited as Rhodococcus through 16S rDNA sequential analysis and belongs to, therefore called after rhodococcus (Rhodococcus sp.) HZF1.
The invention still further relates to the application of described rhodococcus HZF1 in microbiological deterioration 4-fluoro cinnamic acid.
Concrete, described degraded is carried out under 25 ~ 45 DEG C, pH5.0 ~ 9.0, dark condition, generally needs vibration (100 ~ 200 rpm).
Described degraded is carried out under 30 DEG C, pH 7.0,150rpm, dark condition.
Be preferably 200 mg/L of 100 ~ 1500 mg/L(at 4-fluoro cinnamic acid final concentration) minimal medium, add the cell suspension containing rhodococcus HZF1, the dark shaking culture 16 ~ 24h of the condition that is 5.0 ~ 9.0 at 25 ~ 45 DEG C, pH value, can make the quality residual quantity of 4-fluoro cinnamic acid in reaction solution be less than 4%; It is 1 ~ 5 × 10 that the described cell suspension add-on containing rhodococcus HZF1 makes rhodococcus HZF1 final concentration in reaction system
7individual/ml.
In practical application, described bacterial strain conventionally need to be through overactivation and enlarged culturing, and detailed process is as follows:
(1) slant culture: rhodococcus HZF1 is inoculated in to slant medium, cultivates 5 ~ 7 days for 25 ~ 45 DEG C, obtain thalline inclined-plane; The final concentration of described slant medium consists of: yeast powder 10g/L, and peptone 5.0g/L, sodium-chlor 10.0g/L, agar 20.0g/L, solvent is water;
(2) seed culture: picking one transfering loop thalline is seeded in minimal medium from step (1) thalline inclined-plane, cultivates 5 ~ 7 days for 25 ~ 45 DEG C, obtains seed liquor; Described minimal medium final concentration consists of: in every liter of nutrient solution, contain NaCl 1g, K
2hPO
41.5g, KH
2pO
40.5g, (NH4)
2sO
41.5g, MgSO
40.1g, 1ml trace element solution, solvent is water, natural pH value makes after high pressure steam sterilization (121 DEG C, 20min), wherein in every liter of trace element solution, contains MnSO
4﹒ H
2o 0.13
g, ZnCl
20.23g, CuSO
4﹒ H
2o 0.03g, CoCl
2﹒ 6H
2o 0.42g, Na
2moO
4﹒ 2H
2o 0.15g, AlCl
3﹒ 6H
2o 0.05g, solvent is water;
(3) enlarged culturing: the seed liquor that step (2) is obtained is seeded in LB liquid nutrient medium with the inoculum size of volumetric concentration 10 ~ 20%, 30 DEG C, 150rpm vibration are supported to logarithmic phase, obtain bacterium liquid, by centrifugal bacterium liquid, abandon supernatant, the phosphoric acid buffer that precipitation is 7.0 by pH value suspends, and obtains the cell suspension containing rhodococcus HZF1, and this cell suspension can add the degraded for 4-fluoro cinnamic acid in water body or soil; Described LB liquid nutrient medium final concentration consists of: in every liter of cultivation, contain yeast powder 10g, and peptone 5.0g, sodium-chlor 10.0g, solvent is water, natural pH value.
Thalli growth amount of the present invention adopts ultraviolet spectrophotometer to detect, and represents at the absorbance at 600nm place by measuring thalline (being mycetocyte nutrient solution).
The present invention adopts reversed-phased high performace liquid chromatographic to detect the residual quantity of 4-fluoro cinnamic acid in minimal medium.RPLC testing conditions: moving phase is methyl alcohol: 10 mM acetic acid, ammonium acetate buffer solution=50:50(volume ratio), analytical column is Grace Alltima C18 chromatographic column (4.6 × 250mm, 5 μ m), flow velocity is 0.8ml/min, sample size is 10 μ l, and column temperature is 30 DEG C.
Compared with prior art, beneficial effect of the present invention is mainly reflected in:
Rhodococcus HZF1 of the present invention can be applied to by the mode directly adding the degraded of 4-fluoro cinnamic acid in water body and soil, the residual 4-fluoro cinnamic acid on the object such as water body, soil of safely, efficiently, fastly degrading, the microbial inoculum preparation technology who contains this bacterial strain is simple, with low cost, easy to use, there is good application prospect.
(4) brief description of the drawings
Fig. 1 is the Electronic Speculum figure of rhodococcus HZF1 of the present invention;
Fig. 2 is the canonical plotting of 4-fluoro cinnamic acid;
Fig. 3 is rhodococcus HZF1 of the present invention degradation curve figure to 4-fluoro cinnamic acid under differing temps: square (
) be 30 DEG C, circular (
) be 25 DEG C, equilateral triangle (
) be 37 DEG C, del (
) be 45 DEG C;
Fig. 4 is that under different pH, on 4-fluoro cinnamic acid, degraded affects figure to rhodococcus HZF1 of the present invention;
Fig. 5 is the degradation curve figure of the rhodococcus HZF1 of the present invention 4-fluoro cinnamic acid that is 200mg/L to final concentration under pure culture condition;
Fig. 6 is that rhodococcus HZF1 of the present invention is the growth curve chart under 200mg/L pure culture condition at 4-fluoro cinnamic acid final concentration;
Fig. 7 is the degradation curve figure of rhodococcus HZF1 of the present invention to different concns 4-fluoro cinnamic acid.
(5) embodiment
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in this:
Embodiment 1: the screening of bacterial strain and qualification
1) substratum
The preparation of minimal medium: NaCl 1g, K
2hPO
41.5g, KH
2pO
40.5g, (NH
4)
2sO
41.5g, MgSO
40.1g, 1ml trace element solution, distilled water complements to 1000ml, after mixing, stirs, and natural pH value makes after high pressure steam sterilization (121 DEG C, 20min), wherein in every liter of trace element solution, contains MnSO
4﹒ H
2o 0.13
g, ZnCl
20.23g, CuSO
4﹒ H
2o 0.03g, CoCl
2﹒ 6H
2o 0.42g, Na
2moO
4﹒ 2H
2o 0.15g, AlCl
3﹒ 6H
2o 0.05g, complements to 1000ml with distilled water.
Enrichment culture liquid: add 4-fluoro cinnamic acid in minimal medium, the final concentration that makes 4-fluoro cinnamic acid is 500mg/L.
The preparation of LB liquid nutrient medium: yeast powder 10g, peptone 5.0g, sodium-chlor 10.0g, distilled water complements to 1000ml, after mixing, stirs, and natural pH value makes after high pressure steam sterilization (121 DEG C, 20min).
The preparation of LB solid medium: yeast powder 10g, peptone 5.0g, sodium-chlor 10.0g, agar 15.0g, distilled water complements to 1000ml, after mixing, stirs, and natural pH value makes after high pressure steam sterilization (121 DEG C, 20min).
2) strains separation purifying
Mud sample picks up from Hangzhou insecticide factory, get 5ml mud sample and be placed in 250ml Erlenmeyer flask, add 100ml enrichment culture liquid, dark shaking culture (30 DEG C, 150rpm) 1 week, gets the turbid liquid in 5ml upper strata in fresh enrichment culture liquid, continue (30 DEG C of dark shaking culture, 150rpm) 1 week, repeat aforesaid operations process 3 times, each inoculum of cultivating is all taken from the nutrient solution of cultivating gained last time.
The nutrient solution 5ml that gets last cultivation gained carries out gradient dilution (10
-4, 10
-5, 10
-6), the nutrient solution 150 μ l that get after each dilution coat on the LB solid medium flat board containing 500mg/L4-fluoro cinnamic acid, being placed in constant incubator (30 DEG C) cultivates, until growing on flat board after bacterium colony, the each bacterium colony of picking is in containing purifying repeatedly on the LB solid medium flat board of 500mg/L4-fluoro cinnamic acid, until bacterium colony is single, each bacterium colony after purifying is connected to respectively in LB liquid nutrient medium test tube to (30 DEG C of shaking culture, 150rpm) spend the night, cultivate 3d by being connected in enrichment culture liquid after centrifugal cultured bacterium liquid, detect the residual quantity of 4-fluoro cinnamic acid in each enrichment culture liquid by reversed-phased high performace liquid chromatographic, finally screening obtains the bacterial strain of a strain energy efficient degradation 4-fluoro cinnamic acid, called after HZF1.
3) identification of strains
The bacterial strain of above-mentioned acquisition is carried out to morphological specificity and molecular biology identification, and the electromicroscopic photograph of this bacterial strain as shown in Figure 1.The main biological property of this bacterial strain is: thalline is spherical, and atrichia, without gemma, size is about that (m), bacterium colony is smooth for 1.7 ~ 3.2 μ, intermediate projections, edge-diffusion, be light orange, catalase feminine gender, oxidase negative, the methyl red test positive, can utilize beta-cyclodextrin, starch, glucose, polysorbate40, acetate, voges-Proskauer test feminine gender, V.P. reacting positive.The optimum growth conditions of this bacterial strain is pH value 7.0,30 DEG C of temperature.This bacterial strain is accredited as Rhodococcus through 16S rDNA sequential analysis and belongs to, therefore called after rhodococcus (Rhodococcus sp.) HZF1(CCTCC No:M 2012404).
Embodiment 2: the preparation of mycetocyte suspension
(1) slant culture: rhodococcus HZF1 is inoculated in to slant medium, cultivates 6 days for 30 DEG C, obtain thalline inclined-plane; The final concentration of described slant medium consists of: yeast powder 10g, peptone 5.0g, sodium-chlor 10.0g, agar 20.0g, water 1000ml;
(2) seed culture: picking one transfering loop thalline is seeded in minimal medium from step (1) thalline inclined-plane, cultivates 6 days for 30 DEG C, obtains seed liquor; Described minimal medium final concentration forms with embodiment 1;
(3) enlarged culturing: the seed liquor that step (2) is obtained is seeded in LB liquid nutrient medium (100 mL) with the inoculum size of volumetric concentration 10 ~ 20%, 30 DEG C, 150rpm shaking culture are to logarithmic phase, obtain bacterium liquid, by centrifugal bacterium liquid (8000rpm, 5min), abandon supernatant, the phosphoric acid buffer that precipitation is 7.0 by pH value suspends, obtain containing rhodococcus HZF1 cell suspension 100 mL, wherein the rhodococcus HZF1 concentration in cell suspension is 5 × 10
8individual/ml;
Described LB liquid nutrient medium final concentration forms with embodiment 1;
PH is that the formula of the phosphoric acid buffer of 7.0 0.2 mol/L is: get the SODIUM PHOSPHATE, MONOBASIC 39ml of 0.2 mol/L and the Sodium phosphate dibasic 61ml of 0.2mol/L, be settled to 1000ml with ultrapure water, after high pressure steam sterilization (121 DEG C, 20min) and get final product.
Embodiment 3:4-fluoro cinnamic acid degradation experiment
1) detection of cell concentration and 4-fluoro cinnamic acid content in minimal medium:
Thalli growth amount adopts ultraviolet spectrophotometer to detect, and represents at the absorbance at 600nm place by measuring thalline in nutrient solution.
This experiment adopts reversed-phased high performace liquid chromatographic to detect the residual quantity of 4-fluoro cinnamic acid in minimal medium.RPLC testing conditions: moving phase is methyl alcohol: 10 mM acetic acid, ammonium acetate buffer solution=50:50(volume ratio), analytical column is Grace Alltima C18 chromatographic column (4.6 × 250mm, 5 μ m), flow velocity is 0.8ml/min, sample size is 10 μ l, and column temperature is 30 DEG C.
2) 4-fluoro cinnamic acid degradation experiment:
4-fluoro cinnamic acid standard substance are dissolved to the reference liquid that is mixed with 100 mg/L with sterilized water, at reversed-phase liquid chromatography testing standard curve, as shown in Figure 2, typical curve equation is y=4.541x+11.737 to 4-fluoro cinnamic acid typical curve, R
2=0.9951, y is peak area, and x is 4-fluoro cinnamic acid concentration.
A, the impact of differing temps on degraded: get 4 250ml Erlenmeyer flasks, add respectively 100ml minimal medium, (121 DEG C of high pressure steam sterilizations, 20min), add 4-fluoro cinnamic acid, make 4-fluoro cinnamic acid final concentration be 200 mg/L, respectively get mycetocyte suspension 5ml(5 × 10 that embodiment 2 methods obtain
8individual/ml), be inoculated in respectively in this minimal medium, make rhodococcus HZF1 final concentration be about 2.5 × 10
7individual/ml, respectively at 25,30,37,45 DEG C of cultivation shaking tables (pH 7.0,150rpm), timing sampling per hour is measured residual 4-fluoro cinnamic acid concentration, and as shown in Figure 3, show 30 DEG C is the suitableeest degradation temperature to result.
B, the different pH impact on degraded: get 5 250ml Erlenmeyer flasks, add respectively 100ml minimal medium, (121 DEG C of high pressure steam sterilizations, 20min), add 4-fluoro cinnamic acid, make 4-fluoro cinnamic acid final concentration be 200 mg/L, respectively get the mycetocyte suspension 5ml that embodiment 2 methods obtain, be inoculated in respectively in this minimal medium, make rhodococcus HZF1 final concentration be about 2.5 × 10
7individual/ml, regulates respectively pH to 5.0,6.0,7.0,8.0 and 9.0, and under 30 DEG C, 150rpm, shaking table is cultivated, and timing sampling per hour is measured residual 4-fluoro cinnamic acid concentration, and as shown in Figure 4, demonstration pH7.0 is the suitableeest degraded pH to result.
C, the impact of different time on degraded: get 250ml Erlenmeyer flask, add 100ml minimal medium, after high pressure steam sterilization (121 DEG C, 20min), add 4-fluoro cinnamic acid, making 4-fluoro cinnamic acid final concentration is 200 mg/L, and 3 Duplicate Samples are set altogether.Respectively get the mycetocyte suspension 5ml that embodiment 2 methods obtain, be inoculated in respectively in this minimal medium, make rhodococcus HZF1 final concentration be about 2.5 × 10
7individual/ml, arranges 3 parallel laboratory tests that do not contain this bacterial classification accordingly as blank, is then together placed in dark shaking culture in shaking table (30 DEG C, pH7.0,150rpm).Be 0,3,6,9,12,24,36,48,60 at incubation time, timing sampling when 72h, detect the increment of thalline in minimal medium and the residual quantity of 4-fluoro cinnamic acid according to above-mentioned detection method, the results are shown in Figure shown in 5.
As shown in Figure 5, as shown in Figure 6, Fig. 6 can find out OD to the growth curve of thalline to the degradation curve of the 4-fluoro cinnamic acid of bacterial strain of the present invention to 200 mg/L concentration
600be increased to 0.42 from 0.18, illustrate that thalli growth is good, observe Fig. 5, can find, cultivate after 24h, the degradation rate of the 4-fluoro cinnamic acid of 4-fluoro cinnamic acid degradation bacteria of the present invention to 200 mg/L is close to 100%, and the percent hydrolysis of all blanks that do not add bacterium after 24h is all less than 5%.
D, the impact that different concns 4-fluoro cinnamic acid is degraded:
Get 4 250ml Erlenmeyer flasks, add respectively 100ml minimal medium, (121 DEG C of high pressure steam sterilizations, 20min), add 4-fluoro cinnamic acid, make 4-fluoro cinnamic acid final concentration be respectively 200,500,1000,1500 mg/L, respectively get the mycetocyte suspension 5ml that embodiment 2 methods obtain, be inoculated in respectively in this minimal medium, make rhodococcus HZF1 final concentration be about 2.5 × 10
7individual/ml, under 7.0,30 DEG C of pH, 150rpm, shaking table is cultivated, and timing sampling is measured residual 4-fluoro cinnamic acid concentration, and result is as shown in Figure 7.
Bacterial strain of the present invention to concentration be 200 ~ 1500 mg/L 4-fluoro cinnamic acid degradation rate as shown in Figure 7, result shows all have extraordinary degradation capability, and this bacterial classification is novel 4-fluoro cinnamic acid degradation bacteria, therefore, this bacterium has very large promoter action to degradation pathway and the degrading genes of research 4-fluoro cinnamic acid, and the degraded of 4-fluoro cinnamic acid in environment is especially had to certain positive effect to the concentrated reparation of 4-fluoro cinnamic acid.
Claims (4)
1. rhodococcus (Rhodococcus sp.) HZF1, is preserved in Chinese Typical Representative culture collection center, address: China, and Wuhan, Wuhan University, postcode 430072, preservation date is on October 15th, 2012, deposit number is CCTCC No:M 2012404.
2. the application of rhodococcus HZF1 as claimed in claim 1 in microbiological deterioration 4-fluoro cinnamic acid.
3. application as claimed in claim 2, is characterized in that described degraded carries out under 25 ~ 45 DEG C, pH5.0 ~ 9.0, dark condition.
4. application as claimed in claim 3, is characterized in that described degraded carries out under 30 DEG C, pH 7.0,150rpm, dark condition.
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| 4-fluorocinnamic acid biodegradation by a rhodococcus strain;Amorim, C. L. et al.;《5th European Bioremediation Conference》;20110707;1-20 * |
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