CN105296389B - One plant of Benzylpenicillin sodium salt degradation bacteria PC-2 and its application - Google Patents
One plant of Benzylpenicillin sodium salt degradation bacteria PC-2 and its application Download PDFInfo
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- CN105296389B CN105296389B CN201510719262.8A CN201510719262A CN105296389B CN 105296389 B CN105296389 B CN 105296389B CN 201510719262 A CN201510719262 A CN 201510719262A CN 105296389 B CN105296389 B CN 105296389B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The invention belongs to biodegradable process fields, provide the degradation application of one plant of Benzylpenicillin sodium salt degradation bacteria and its separation identification and the degradation bacteria on Benzylpenicillin sodium salt.Benzylpenicillin sodium salt degradation Pseudomonas chelates Coccus, and Classification And Nomenclature chelates coccus Chelatococcus sp., and deposit number is CGMCC No.1.15283, and China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on June 11st, 2015.Degradation characteristic is studies have shown that the degradation rate for the Benzylpenicillin sodium salt that shaken cultivation 12d, PC 2 is 200ppm to initial concentration under conditions of 37 DEG C, 150r/min is 53.5%.When glucose is carbon source, peptone is nitrogen source, degradation bacteria inoculum concentration is 14%, Benzylpenicillin sodium salt initial concentration is 100ppm, pH value range 6.0~7.0 when degradation rate highest.The Benzylpenicillin sodium salt degradation bacteria has good degradation characteristic to Benzylpenicillin sodium salt, and certain reference frame is provided for bioanalysis degradation Penicillin Residues.
Description
Technical field
The invention belongs to biodegradable process fields, are related to one plant of Benzylpenicillin sodium salt degradation bacteria and its separation identification, and should
Degradation application of the degradation bacteria on Benzylpenicillin sodium salt.
Background technology
Penicillin mushroom dregs are the main waste materials in China's Fermentation Industry production, according to preresearch estimates, China in 2009
About 1,300,000 tons of all kinds of antibiotic bacterium dregs generated.The departments such as the Ministry of Agriculture, the Ministry of Public Health, National Drug Administration in 2002 handle
Antibiotic bacterium dregs are classified as in the types of drugs catalogue for forbidding using in feed and animal drinking water；2008, State Ministry of Environmental Protection
Antibiotic bacterium dregs are included in again《National Hazard waste register》In.For the reality such as antibiotic bacterium dregs yield is big, intractability is big
Problem, and《Pharmaceuticals industry pollution prevention technique policy》What is proposed in (exposure draft) " encourages exploitation fermentation bacteria residue in life
How reutilization technology, innoxious process for treating, comprehensive utilization technique in production. art " policy and suggestion, realize antibiotic bacterium dregs
Rational and efficient use and safe handling become the problem of China environmental protection and pharmaceutical industries sustainable development.
Antibiotic bacterium dregs composting technology can not only efficiently use a large amount of protein, organic matter, organic acids in bacteria residue, moreover it is possible to drop
Remaining antibiotic in bacteria residue is solved, therefore is the research hotspot of antibiotic bacterium Slag treatment disposal technology.If not in view of compost treatment
Antibiotic residue can be completely removed, certain Ecological Environment Risk can be led to after composting production application, some scholars start
Microbe inoculation microbial inoculum in composting process, to improve the degradation rate of compost material.
Screening, the separation of Degradation of Antibiotics bacterium are the key that Biochemical method antibiotic pollutants.So far, both at home and abroad
Some researchers have screened more plants of Degradation of Antibiotics bacterium, but penicillin degradation bacteria is less, and the degradation bacteria that this research is related to is
Chelating Coccus (Chelatococcus sp.) bacterium of the Benzylpenicillin sodium salt that can degrade found for the first time.
Invention content
In order to overcome antibiotic in current antibiotic bacterium dregs to be difficult to effectively decompose, cause penicillin mushroom dregs efficiently sharp
With, the problems such as generating a large amount of pollution, provide one plant can efficient-decomposition Benzylpenicillin sodium salt degradation bacteria and the degradation bacteria it is green in degradation
Application in terms of mycin sodium.
The purpose of the present invention is achieved through the following technical solutions
1. the present invention provides one plant of Benzylpenicillin sodium salt degradation bacteria PC-2, Benzylpenicillin sodium salt degradation Pseudomonas chelates Coccus, classification
Name chelating coccus Chelatococcus sp., deposit number was CGMCC No.1.15283, in preservation on June 11 in 2015
In China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Preservation address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica.
The Benzylpenicillin sodium salt degradation bacteria PC-2 biological properties are as follows：PC-2 is rounded on LB solid mediums, intermediate
Protrusion, milky, surface is smooth, opaque, neat in edge, and colony diameter is about 1~2mm.It is seen under transmission electron microscope
It examines, PC-2 is rod-short, and 1.96 0.92 μm of μ ms hold raw flagellum.
2. the present invention also provides the method for identification Benzylpenicillin sodium salt degradation bacteria PC-2, this method with primers F-primer 27F and
R-primer 1492R treat detection bacterium genome and carry out PCR amplification, obtain the amplified production of 1475bp, illustrate that this is to be detected
Bacterium is Benzylpenicillin sodium salt degradation bacteria PC-2；
Wherein, the sequence of primers F-primer 27F is：5′-AGAGTTTGATCCTGGCTCAG-3′；R-
The sequence of primer1492R is：5′-TACGGCTACCTTGTTACGACTT-3′.
The amplified production of 1475bp is 16S rDNA sequence PCR analysis results.
3. the present invention also provides the methods of separating penicillin sodium degradation bacteria PC-2, including：(1) penicillin mushroom dregs and pig manure are taken
Hybrid composting sample, be added in triangular flask containing distilled water, 36 DEG C -38 DEG C, 130r/min-160r/min shakes on shaking table
Sample is set to be sufficiently mixed with water；Wherein, in the hybrid composting sample of penicillin mushroom dregs and pig manure bacteria residue and compost mass ratio
It is 1:2；The ratio of hybrid composting and distilled water is 1:8-1:10g/mL
(2) it when mixed liquor being diluted to concentration using gradient dilution method and being about 0.1~1.0ppm, is pipetted respectively with liquid-transfering gun
The sample of 200 μ L is applied to Benzylpenicillin sodium salt and adds on the LB solid mediums of a concentration of 1000ppm in 36-37 DEG C of constant temperature incubation
2d-3d, after culture medium grows bacterium colony, the scribing line of picking single bacterium colony obtains single strain after being separately cultured repeatedly.
4. the present invention also provides application of the Benzylpenicillin sodium salt degradation bacteria described in above-mentioned 1 in Penicillin Residues degradation, this is answered
It is 100-1200ppm's with by the way that penicillin degradation bacteria PC-2 is inoculated in Benzylpenicillin sodium salt initial concentration by inoculum concentration 2%-18%
In Penicillin Residues object, carbon source, nitrogen source are added thereto, adjustment pH value is 6-10；Wherein, carbon source includes glucose, maltose
With one or more mixtures of sucrose, a concentration of 0.3%-0.5%；Nitrogen source includes one kind of peptone, yeast extract and urea
Or a variety of mixtures, a concentration of 0.3%-0.5%.
5. application of the above-mentioned 4 Benzylpenicillin sodium salt degradation bacterias provided in Penicillin Residues degradation, wherein the inoculum concentration is
14%, Benzylpenicillin sodium salt initial concentration is 100ppm, the grape that the carbon source being added into Penicillin Residues object is a concentration of 0.4%
Sugar, the peptone that nitrogen source is a concentration of 0.4%, adjustment pH ranges are 6.0~7.0.
6. the application described in above-mentioned 4 or 5, wherein penicillin degradation bacteria PC-2 is derived from exponential phase, is first passed through before inoculation
Centrifugation to be crossed, is then eluted with phosphate buffer solution, the phosphate buffer solution is 0.2mol/L NaH2PO4 39.0mL,
7.0,121 DEG C of sterilizing 20min of 0.2mol/L Na2HPO461.0mL, pH.
7. the present invention also provides a kind of Benzylpenicillin sodium salt degradation agent, which includes Benzylpenicillin sodium salt degradation bacteria PC-2.
8. application of the above-mentioned 7 Benzylpenicillin sodium salt degradation agents provided in Benzylpenicillin sodium salt degradation, in use, being dropped according to penicillin
Solution bacterium PC-2 inoculum concentrations 2%-18% is inoculated in the Penicillin Residues object that Benzylpenicillin sodium salt initial concentration is 100-1200ppm, to
Carbon source, nitrogen source is wherein added, adjustment pH value is 6-10；Wherein, carbon source includes the one or more of glucose, maltose and sucrose
Mixture, a concentration of 0.3%-0.5%；Nitrogen source includes one or more mixtures of peptone, yeast extract and urea, a concentration of
0.3%-0.5%.
9. the application described in above-mentioned 8, wherein the inoculum concentration is 14%, and Benzylpenicillin sodium salt initial concentration is 100ppm, Xiang Qing
The glucose that the carbon source being added in mycin residue is a concentration of 0.4%, the peptone that nitrogen source is a concentration of 0.4% adjust pH
Range is 6.0~7.0.
Advantageous effect：
1. the degradation bacteria that research is related to is the chelating Coccus of the Benzylpenicillin sodium salt that can degrade found for the first time
(Chelatococcus sp.) bacterium, has filled up the blank of current research.
2. by response parameter during optimization degradation bacteria degradation Benzylpenicillin sodium salt, Benzylpenicillin sodium salt degradation bacteria PC-2 degradation moulds
The degradation rate of plain sodium can reach 99.32%, and the degradation rate compared to general 10% is significantly improved, and illustrate that the present invention carries
The penicillin degradation bacteria PC-2 of confession is a kind of high efficiency degradation bacteria.It can be a wide range of using Benzylpenicillin sodium salt degradation agent made of this bacterium
In being produced applied to Fermentation Industry, environmental pollution can be substantially reduced, improves business efficiency.
Description of the drawings
Fig. 1 is colonial morphologies of the PC-2 on LB tablets
Fig. 2 is PC-2 transmission electron microscope pictures
Fig. 3 is PC-2 scanning electron microscopic picture
Fig. 4 is PC-2 phylogenetic tree pictures
Fig. 5 is Benzylpenicillin sodium salt solution canonical plotting
Fig. 6 is PC-2 growth curve pictures
Fig. 7 is PC-2 degradation curve pictures
Fig. 8 is PC-2 degradation kinetics equations of linear regression
Fig. 9 is degradation effect influence diagram of the carbon source to PC-2 degradation Benzylpenicillin sodium salts
Figure 10 is degradation effect influence diagram of the nitrogen source to PC-2 degradation Benzylpenicillin sodium salts
Figure 11 is degradation effect influence diagram of the inoculum concentration to PC-2 degradation Benzylpenicillin sodium salts
Figure 12 is degradation effect influence diagram of the concentration of substrate to PC-2 degradation Benzylpenicillin sodium salts
Figure 13 is degradation effect influence diagram of the pH value to PC-2 degradation Benzylpenicillin sodium salts
Specific implementation mode
With reference to embodiment, the present invention will be further described, and the experiment side of actual conditions is not specified in the following example
Method, usually according to the known approaches of this field.
LB culture mediums：Tryptone 10g, yeast extract 5g, NaCl 10g, distilled water 1000mL, 121 DEG C of sterilizings
20min.LB solid mediums add 15g agar powders.
Basal medium：(NH4)2SO42g, NaH2PO40.5g, K2HPO40.5g, MgSO40.2g, distilled water are settled to
1000mL, 121 DEG C of sterilizing 20min.
Above two culture medium is respectively used to the drafting of degradation bacteria PC-2 growth curves and degradation curve.
Phosphate buffer solution：0.2mol/L NaH2PO439.0mL, 0.2mol/L Na2HPO461.0mL, pH7.0,
121 DEG C of sterilizing 20min.
Major experimental instrument and reagent：Bio-incubator (Thermo)；Centrifuge (eppendrof, Centrifuge
5415R)；Constant-temperature table (Thermo, MAXQ 4000)；High performance liquid chromatograph Waters 2695 (Waters, US).
Acetonitrile (chromatographically pure, German Merck companies)；Benzylpenicillin sodium salt standard items (>98%, Sigma Co., USA)；Formic acid
(analyzing pure, Shanghai Ling Feng chemical reagent Co., Ltd)；Experimental water is Milli Q water (conductivity 18.2M Ω cm).PCR
Reagent is purchased from TaKaRa companies.
Embodiment 1
The screening and identification of Benzylpenicillin sodium salt degradation bacteria PC-2
(1) separation screening of Benzylpenicillin sodium salt degradation bacteria PC-2
Take the hybrid reactor of 10g penicillin mushroom dregs (the Hebei pharmacy of Hebei Shijiazhuang) and pig manure (the Nanjing towns Heng Xi)
Fertile sample (compost place is Nanjing Agricultural University Of Nanjing decorated archway), the mass ratio of bacteria residue and pig manure is 1:2), add
It is added in the triangular flask of 250mL of the 90mL containing distilled water, 37 DEG C, 150r/min shakes on shaking table keeps sample fully mixed with water
It closes.When mixed liquor being diluted to concentration using gradient dilution method being about 0.1~1.0ppm, pipette 200 μ L's respectively with liquid-transfering gun
Sample is applied on the LB solid mediums of 1000ppm Benzylpenicillin sodium salt additive amounts, in 37 DEG C of constant temperature incubation 2d, waits for that culture medium is grown
After bacterium colony, the scribing line of picking single bacterium colony obtains single strain after being separately cultured about 4 times.By obtain it is purebred be transferred on the test tube slants LB,
37 DEG C of constant temperature incubation 48h are put into 4 DEG C of refrigerators and preserve.
(2) morphology of Benzylpenicillin sodium salt degradation bacteria PC-2 and molecules identification
Cell morphological characteristic is observed with transmission electron microscope (TEM), bacterial strain identification uses morphologic observation and 16S rDNA
Sequence PCR analysis methods.
Bacterial strain PC-2 is rounded on LB solid mediums, and intermediate projections, milky, surface is smooth, opaque, and edge is whole
Together, colony diameter is about 1~2mm (as shown in Figure 1).It being observed under transmission and scanning electron microscope, PC-2 is rod-short,
1.96 0.92 μm of μ ms hold raw flagellum (as shown in Figure 2,3).
The primer of 16S rDNA amplifications is F-primer 27F：5′-AGAGTTTGATCCTGGCTCAG-3′；R-primer
1492R：5′-TACGGCTACCTTGTTACGACTT-3′.Amplified production is sequenced by Sangon Biotech (Shanghai) Co., Ltd.,
It is compared using blast program and the known array in NCBI.
PC-2 and Chelatococcus daeguensis (T) K106 and Chelatococcus sambhunathii (T)
HT4 clusters according to morphological observation and combine document report in a systematic growth branch, sequence homology 99% or more,
Preliminary Identification PC-2 belongs to chelating Coccus (Chelatococcus sp.).
Embodiment 2
Benzylpenicillin sodium salt degradation bacteria PC-2 degradation characteristics
(1) drafting of Benzylpenicillin sodium salt solution standard curve
Benzylpenicillin sodium salt concentration is quantitative determined using Ultra Performance Liquid Chromatography instrument (Waters 2695, the U.S.).Chromatographic condition：
Mobile phase：0.1% aqueous formic acid-acetonitrile (V:V=50:50)；Flow velocity：1mL/min；Detector is PDA UV detector；
Wave-length coverage 190nm~400nm；Column temperature：30℃；20 μ L of sample size；Appearance time is set as 6min.
Compound concentration is respectively the Benzylpenicillin sodium salt working solution of 0.5ppm, 1ppm, 2ppm, 4ppm, 8ppm, then accurate respectively
It draws standard working solution 1mL sample introductions under the chromatographic condition of setting to measure, with the mass concentration of Benzylpenicillin sodium salt solution for horizontal seat
Mark draws standard curve (as shown in Figure 5) by ordinate of its peak area.
(2) degradation bacteria PC-2 growth curves measure
The degradation bacteria PC-2 placements being stored in 4 DEG C of refrigerators are restored a few minutes at room temperature, then picking mycelium turns
It is connected on new slant medium and is cultivated.It is linked into again from picking thalline in new slant strains equipped with 100mL liquid
In the triangular flask (250mL) of LB culture mediums, at 37 DEG C, seed liquor is made in shaken cultivation 36h under the conditions of 150r/min.By seed
Liquid is inoculated into according to 10% inoculum concentration in the triangular flask equipped with 100mL LB liquid mediums (500mL), in 37 DEG C, 150r/
Shaken cultivation 72h under the conditions of min takes a sample every 3h, and microplate reader is used in combination to measure OD600Value determines the increment of thalline.If
The culture medium not being inoculated with is blank control.
As can be seen that 0-6h is the growth adaptation phase from the growth curve of PC-2,6-36h is logarithmic phase, is entered after 36h steady
Periodically (as shown in Figure 6).
(3) the degradation kinetics experiment of degradation bacteria PC-2 degradation efficiencies and residual Benzylpenicillin sodium salt
Exponential phase bacterium solution is centrifuged into (rotating speed should not be too large, about 3000r/min) 10min, phosphate buffer solution leaching
After washing about 3 times, it is inoculated in by 10% in the basal medium that Benzylpenicillin sodium salt initial concentration is 200ppm.37 DEG C, 150r/min shakes
Culture is swung, is sampled every 3h, Benzylpenicillin sodium salt concentration is measured.Experiment sets three repetitions, and the culture medium to be not added with Benzylpenicillin sodium salt is opposed
According to.
The result shows that the degradation rate of Benzylpenicillin sodium salt increases with the extension of incubation time, when to 12d, Benzylpenicillin sodium salt is dense
It is 53.5% (as shown in Figure 7) that degree is reduced to 93ppm, degradation rate by initial 200ppm.At any time according to Benzylpenicillin sodium salt concentration
Variation, obtain the degradation kinetics equation of linear regression of Benzylpenicillin sodium salt：Y=-9.008x+203.6, R2=0.993, see Fig. 8.
(4) influence of the medium component to Benzylpenicillin sodium salt degradation efficiency
(1) influence of the carbon source to Benzylpenicillin sodium salt degradation efficiency
0.4% glucose, maltose and sucrose are separately added into basal medium.After accessing 10% seed liquor, 37
DEG C, 150r/min shaken cultivation 48h measure Benzylpenicillin sodium salt content, calculate Benzylpenicillin sodium salt degradation rate, determine best carbon source.It is real
It tests and sets 3 repetitions, as a result take its average value.
When PC-2 is cultivated using Benzylpenicillin sodium salt as sole carbon source (control group), Benzylpenicillin sodium salt degradation rate is only
11.56%.Carbon source is added, the degradation rate of Benzylpenicillin sodium salt can be improved.Glucose is added in basal medium, Benzylpenicillin sodium salt
Sucrose and maltose is followed by added in degradation rate highest.Therefore, optimum carbon source is glucose.(as shown in Figure 9)
(2) influence of the nitrogen source to Benzylpenicillin sodium salt degradation efficiency
0.4% peptone, yeast extract and urea are separately added into basal medium.After accessing 10% seed liquor, 37
DEG C, 150r/min shaken cultivation 48h measure Benzylpenicillin sodium salt content, calculate Benzylpenicillin sodium salt degradation rate, determine best nitrogen source.It is real
It tests and sets 3 repetitions, as a result take its average value.
From the point of view of degradation effect, peptone>Yeast extract>Urea>Control.Therefore, select peptone for best nitrogen
Source.(as shown in Figure 10)
(5) influence of the chemical factors to Benzylpenicillin sodium salt degradation efficiency
(1) influence of the inoculum concentration of degradation bacteria PC-2 to Benzylpenicillin sodium salt degradation efficiency
Exponential phase bacterium solution is centrifuged, after phosphate buffer solution elution, presses 2%, 6%, 10%, 14% and respectively
18% inoculum concentration accesses in the culture medium optimized that Benzylpenicillin sodium salt initial concentration is 200ppm.37 DEG C, 150r/min shakes
Culture 6h is swung, Benzylpenicillin sodium salt content is measured, Benzylpenicillin sodium salt degradation rate is calculated, determines best inoculum concentration.Experiment sets 3 repetitions,
As a result its average value is taken.
The result shows that when inoculum concentration is relatively low, bacterial strain PC-2 is to the degradation effect of Benzylpenicillin sodium salt with the increase of inoculum concentration one
Determine to increase in range, reach highest when inoculum concentration is 14%, is 93.78%.After inoculum concentration is more than 14%, PC-2 pairs of bacterial strain
The degradation effect of Benzylpenicillin sodium salt declines, and therefore, selects 14% as optimum inoculation amount.(as shown in figure 11)
(2) influence of the substrate Benzylpenicillin sodium salt concentration to Benzylpenicillin sodium salt degradation efficiency
Exponential phase bacterium solution is centrifuged, after phosphate buffer solution elution, penicillin is accessed to by 10% inoculum concentration
Sodium initial concentration is respectively in the culture medium of 100,200,400,800 and 1200ppm optimized.37 DEG C, 150r/min oscillations
6h is cultivated, bacterium solution is taken to centrifuge, measures Benzylpenicillin sodium salt content, Benzylpenicillin sodium salt degradation rate is calculated, determines best concentration of substrate.Experiment
If 3 repetitions, as a result take its average value.
The result shows that with the increase of concentration of substrate, the degradation rate of Benzylpenicillin sodium salt continuously decreases, and is reduced to from 99.32%
52.52%, therefore best concentration of substrate is 100ppm.(as shown in figure 12)
(3) influence of the degradation pH value to Benzylpenicillin sodium salt degradation efficiency
The initial pH value that Optimal Medium is adjusted with 1mol/L hydrochloric acid and NaOH is 6.0,7.0,8.0,9.0,10.0, respectively
After accessing 10% seed liquor, 37 DEG C, 150r/min shaken cultivation 6h, Benzylpenicillin sodium salt content is measured, calculates Benzylpenicillin sodium salt degradation rate,
Determine best pH value.Experiment sets 3 repetitions, as a result takes its average value.
The result shows that Benzylpenicillin sodium salt is more stable when pH is 6.0 and 7.0, it is not inoculated with degradation bacteria, the degradation after 6h
Rate is respectively 19.93% and 26.72%.After being inoculated with degradation bacteria PC-2, degradation rate difference of the Benzylpenicillin sodium salt in pH 6.0 and 7.0
Reach 74.05% and 75.9%, shows in 6.0~7.0 ranges of pH, PC-2 plays an important role to the degradation of Benzylpenicillin sodium salt.
(as shown in figure 13)
It is recognised that the illustrative embodiments that above-described embodiment uses only for illustrating inventive principle, however this hair
Bright to be not limited only to this, those skilled in the art can make various improvement and change in the case where not departing from real situation of the present invention, this
A little improvement and change also belong to protection scope of the present invention.
Claims (8)
1. one plant of Benzylpenicillin sodium salt degradation bacteria PC-2, it is characterised in that：Benzylpenicillin sodium salt degradation Pseudomonas chelates Coccus, Classification And Nomenclature
PC-2, deposit number CGMCC1.15283 were preserved in China Committee for Culture Collection of Microorganisms on May 28th, 2015
Common micro-organisms center.
2. a kind of method of Benzylpenicillin sodium salt degradation bacteria PC-2 described in identification claim 1, it is characterised in that：With primers F-primer
27F and R-primer 1492R treat detection bacterium genome and carry out PCR amplification, obtain the amplified production of 1475bp, illustrate this
Bacterium to be detected is Benzylpenicillin sodium salt degradation bacteria PC-2；
Wherein, the sequence of primers F-primer 27F is：5′-AGAGTTTGATCCTGGCTCAG-3′；R-primer 1492R's
Sequence is：5′-TACGGCTACCTTGTTACGACTT-3′.
3. applications of the Benzylpenicillin sodium salt degradation bacteria PC-2 described in claim 1 in Penicillin Residues degradation, it is characterised in that：It will
Penicillin degradation bacteria PC-2 by inoculum concentration 2%-18% be inoculated in Benzylpenicillin sodium salt initial concentration be 100-1200ppm penicillin it is residual
It stays in object, carbon source, nitrogen source is added thereto, adjustment pH value is 6-10；Wherein, carbon source includes glucose, maltose and sucrose
One or more mixtures, a concentration of 0.3%-0.5%；Nitrogen source includes the one or more mixed of peptone, yeast extract and urea
Close object, a concentration of 0.3%-0.5%.
4. application according to claim 3, it is characterised in that：The inoculum concentration is 14%, and Benzylpenicillin sodium salt initial concentration is
100ppm, the glucose that the carbon source being added into Penicillin Residues object is a concentration of 0.4%, the egg that nitrogen source is a concentration of 0.4%
White peptone, adjustment pH ranges are 6.0~7.0.
5. application according to claim 3 or 4, it is characterised in that：Penicillin degradation bacteria PC-2 is derived from exponential phase,
Centrifugation is first passed through before inoculation, is then eluted with phosphate buffer solution, and the phosphate buffer solution is 0.2mol/L NaH2PO4
39.0mL, 0.2mol/L Na2HPO47.0,121 DEG C of sterilizing 20min of 61.0mL, pH.
6. a kind of Benzylpenicillin sodium salt degradation agent, it is characterised in that：The degradation agent includes Benzylpenicillin sodium salt degradation bacteria described in claim 1
PC-2。
7. application of the Benzylpenicillin sodium salt degradation agent in Benzylpenicillin sodium salt degradation described in claim 6, it is characterised in that：In use,
It is inoculated in the penicillin that Benzylpenicillin sodium salt initial concentration is 100-1200ppm according to penicillin degradation bacteria PC-2 inoculum concentrations 2%-18%
In residue, carbon source, nitrogen source are added thereto, adjustment pH value is 6-10；Wherein, carbon source includes glucose, maltose and sucrose
One or more mixtures, a concentration of 0.3%-0.5%；Nitrogen source includes the one or more of peptone, yeast extract and urea
Mixture, a concentration of 0.3%-0.5%.
8. application according to claim 7, it is characterised in that：The inoculum concentration is 14%, and Benzylpenicillin sodium salt initial concentration is
100ppm, the glucose that the carbon source being added into Penicillin Residues object is a concentration of 0.4%, the egg that nitrogen source is a concentration of 0.4%
White peptone, adjustment pH ranges are 6.0~7.0.
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| CN105779339B (en) * | 2016-03-25 | 2019-06-14 | 福建师范大学 | A kind of preparation and its application of aminoglycoside antibiotics mixed bacteria of degradation |
| CN105733989B (en) * | 2016-03-25 | 2019-06-14 | 福建师范大学 | The separation method of aminoglycoside antibiotics degradation bacteria FN-A3 a kind of and its application |
| CN112760262B (en) * | 2021-01-29 | 2022-05-17 | 浙江农林大学 | Erythromycin degradation composite microbial inoculum and preparation method and application thereof |
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