CN106591194B - The variovorax of degradable penicillin, cell fraction and combinations thereof - Google Patents
The variovorax of degradable penicillin, cell fraction and combinations thereof Download PDFInfo
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- CN106591194B CN106591194B CN201611223442.8A CN201611223442A CN106591194B CN 106591194 B CN106591194 B CN 106591194B CN 201611223442 A CN201611223442 A CN 201611223442A CN 106591194 B CN106591194 B CN 106591194B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/26—Organic substances containing nitrogen or phosphorus
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
Abstract
The present invention relates to a kind of variovoraxs of degradable penicillin, cell fraction and combinations thereof, and the variovorax is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center with deposit number CGMCC No.12495.The variovorax of degradable penicillin provided by the invention, cell fraction and combinations thereof, the mycelia that can give up to penicillin carry out harmless treatment, have the advantages that environmentally protective, energy saving, saving cost.
Description
Technical field
The present invention relates to field of microbial culture technology, and in particular to a kind of variovorax of degradable penicillin, cell grade
Point and combinations thereof.
Background technique
Penicillin (Benzylpenicillin) is the general name of a family antibiotic, it is by inhibiting bacteria cell wall tetrapeptide side
Chain and pentapeptide cross-bridge in conjunction with and the synthesis of block cell wall and play bactericidal effect, belong to wide spectrum anti-infectives.
It is raw material that starch, corn pulp, beancake powder etc. are mainly used in the production of industrial penicillin at present, through penicillium chrysogenum
It ferments, extract, purify.For the fermentation liquid after being separated by solid-liquid separation and obtaining benzylpenicillin sodium solution, remaining solid slag is useless
Mycelia.Penicillin gives up mycelia main component as crude protein, cellulose and crude fat etc., wherein also still have a small amount of Penicillin Residues,
Its residual content is about 0.2g/kg-0.5g/kg.Due to remaining the stable in physicochemical property of penicillin in mycelia, there is for a long time residual
The property stayed, bioconcentration easily cause the various security risks such as biology drug resistance after entering biological chain.According to current international and
Domestic law regulation, the useless mycelia of penicillin can only be handled according to solid dangerous waste standard, can not only be made in this way to environment
At secondary pollution, while it will cause the waste of grain resource and the raising of relevant enterprise production cost.What the country generated every year
Penicillin mushroom dregs can reach 100,000 tons or more, it is seen then that there is an urgent need to develop a kind of the practical of the useless mycelia harmless treatment of penicillin
New technology.
However, the method for existing harmless treatment penicillin mycelia mainly has a hot steaming method, soda acid edman degradation Edman, these methods or
Person's energy consumption is big, or causes secondary pollution, and can generate unpleasant exhaust gas.How using environmentally protective means to being remained in bacteria residue
Penicillin carries out appropriate harmless treatment and is efficiently used, and becomes the technical problem of relevant industries.
Summary of the invention
To solve the deficiencies in the prior art, the present invention provides the bacterial strain KDSPL- of one plant of energy degradation selectivity penicillin
04, to lay a good foundation for the useless mycelia harmless biological processing technique of exploitation penicillin.KDSPL-04 bacterial strain is variovorax, should
Variovorax is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms with deposit number CGMCC No.12495
The heart.
Wherein, the colony characteristics that the variovorax is formed on enriched medium are as follows: circle, milk yellow, protrusion, surface light
Sliding, opaque, neat in edge；
Thallus feature are as follows: Gram-negative, thallus is in the shape of a rod, size between 0.9-2.3 μm of 0.4-0.5 μ m, it is single or
Arrangement in pairs.
Wherein, the L- pyrrolidinyl arylamine enzyme, glutamy arylamine enzyme pNA, gamma glutamyltransferase, L- of the variovorax
Proline arylamine enzyme, tyrosine arylamine enzyme, urease, ellman's reagent experiment are positive；Lipase, phosphatase, ornithine decarboxylation
Enzyme, lysine depickling enzyme, histidine assimilation experiment are negative；It is PEARLITOL 25C, D-Glucose, D-MANNOSE, D-Maltose, ancient
Sugared fermenting experiment is negative.
Wherein, the variovorax can be grown by sole carbon source of penicillin.
Wherein, whole base sequences of the variovorax 16S rDNA are as shown in SEQUENCE LISTING.
Invention additionally provides a kind of variovorax of degradable penicillin, greedy bitten by the way that mutagenesis or genetic transformation are above-mentioned
Bacterium obtains.
Wherein, whole base sequences of the variovorax 16S rDNA are as shown in SEQUENCE LISTING.
Invention also provides a kind of cell fractions of degradable penicillin, comprising: from the culture of above-mentioned variovorax
Obtained in DNA prepared product or cell wall preparation, the culture solution obtained in the above-mentioned variovorax, the supernatant of these culture solutions
The culture solution of the cell grade of liquid, the fraction of the supernatant and above-mentioned variovorax.
The present invention also provides a kind of compositions, and it includes above-mentioned variovorax and/or above-mentioned cell fractions.
The variovorax of degradable penicillin provided by the invention, cell fraction and combinations thereof, can give up to penicillin mycelia
Harmless treatment is carried out, has the advantages that environmentally protective, energy saving, saving cost.
Specific embodiment
In order to have further understanding to technical solution of the present invention and beneficial effect, the following detailed description of of the invention
Technical solution and its beneficial effect of generation.
It is variovorax (Variovorax sp.) the present invention provides a kind of classification naming of degradable penicillin, this, which is coveted, bites
It is general that bacterium with deposit number CGMCC No.12495 is preserved in China Committee for Culture Collection of Microorganisms on May 24th, 2016
Logical microorganism center, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, which is KDSPL-04.
Preferably, the colony characteristics that the variovorax is formed on enriched medium are as follows: circle, milk yellow, protrusion, surface
Smooth, opaque, neat in edge；
Thallus feature are as follows: Gram-negative, thallus is in the shape of a rod, size between 0.9-2.3 μm of 0.4-0.5 μ m, it is single or
Arrangement in pairs.
Preferably, the L- pyrrolidinyl arylamine enzyme, glutamy arylamine enzyme pNA, gamma glutamyltransferase of the variovorax,
L-PROLINE arylamine enzyme, tyrosine arylamine enzyme, urease, ELLMAN reagent are positive；Lipase, ornithine decarboxylase, relies phosphatase
Propylhomoserin depickling enzyme, histidine assimilation are negative；PEARLITOL 25C, D-Glucose, D-MANNOSE, D-Maltose, ancient sugar fermentation are negative.
Preferably, the variovorax can be grown by sole carbon source of penicillin.
Preferably, whole base sequences of the variovorax 16S rDNA are as shown in SEQUENCE LISTING.
The variovorax (Variovorax sp.) of degradable penicillin provided by the invention, with efficient degradation penicillin
Ability, the penicillin in mycelium that can be generated to fermentation industry are effectively degraded, to eliminate in the useless mycelia of penicillin
Antibiotic residue, the harmless treatment for the bacteria residue that can be used in penicillin industrial production fermenting, provides for the harmless treatment of mycelia
Effective approach.
Invention additionally provides a kind of variovorax of degradable penicillin, greedy bitten by the way that mutagenesis or genetic transformation are above-mentioned
Bacterium obtains.Specifically, the present invention is by obtaining another degradation mould to the progress mutagenesis of above-mentioned variovorax or genetic transformation screening
The mutation variovorax of element, the bacterial strain at least remain the characteristic of the degradable penicillin of opportunistic pathogen strain, the variovorax bacterial strain packet of the mutation
Include the mutation variovorax bacterial strain obtained by former variovorax bacterial strain by insertion mutation, integration mutation.
Preferably, whole base sequences of the variovorax 16S rDNA are as shown in SEQUENCE LISTING.
Invention also provides a kind of cell fractions of degradable penicillin, comprising: (including former greedy from above-mentioned variovorax
Phagocytosis and the mutation variovorax obtained through mutagenesis or genetic transformation) culture obtained in DNA prepared product or cell wall system
Standby object, the culture solution obtained in the above-mentioned variovorax, the supernatant of these culture solutions and above-mentioned greedy are bitten at the fraction of the supernatant
The culture solution of the cell grade of bacterium.
The present invention also provides a kind of compositions, and it includes above-mentioned variovorax and/or above-mentioned cell fractions.
Variovorax provided by the invention can specifically be screened by following methods, be identified, and measure it to penicillin
Degradation capability and gene order:
Embodiment 1: screening, identification, degradation capability and the sequencing of variovorax bacterial strain
Step 1: sludge 10g is acquired from certain antibiotic enterprise periphery, it is inorganic to be inoculated into the 100mL containing 30mg penicillin
In salt culture medium, in 30 DEG C, 160rpm shaking table shake culture.It is primary that culture starts switching in latter every 7 days, with 10% (V/ when switching
V inoculum concentration) is transferred in the culture medium containing penicillin of higher concentration, penicillin containing concentration be followed successively by 10mg/L,
30mg/L, 60mg/L, 90mg/L, 120mg/L, and so on, until 900mg/L, obtains the enrichment containing thallus of various concentration
Culture solution.
Wherein, the composition and ratio of the minimal medium in the present invention are (g/L): K2HPO41.6g, KH2PO4
0.4g, MgSO4·7H2O 0.2g, CaCl2·2H2O 0.025g, FeCl3·6H2O 0.0023g, NH4NO30.5g, agar
20.0g adjusts pH value to 7.0,121 DEG C of high pressure sterilization 30min with 1mol/L HCl or NaOH.
The composition and ratio of the enriched medium of a certain concentration in the present invention are (g/L): K2HPO41.6g, KH2PO4
0.4g, MgSO4·7H2O 0.2g, CaCl2·2H2O 0.025g, FeCl3·6H2O 0.0023, NH4NO30.5g, yeast extract
0.5g, glucose 1g, peptone 1g, agar 20.0g adjust pH value to 7.0,121 DEG C of high pressures with 1mol/L HCl or NaOH and go out
Bacterium 30min.
Step 2: take in 0.1mL step 1 that be applied to the accordingly selection containing concentration containing germy enrichment culture liquid flat
On plate, cultivated 72 hours at 30 DEG C.Obtain that one plant of speed of growth is most fast and the bacterial strain of obvious transparent circle occurs in periphery of bacterial colonies, by this
In strain inoculated to the minimal medium containing 30mg/L penicillin, periphery of bacterial colonies may occur in which clearly transparent circle, by the bacterium
Strain number is KDSPL-04, variovorax as provided by the invention.
The Main Biological of KDSPL-04 bacterial strain are as follows: the colony characteristics that the bacterial strain is formed on enriched medium are as follows:
Circle, milk yellow, protrusion, surface be smooth, opaque, neat in edge；
The thallus feature of the bacterial strain are as follows: Gram-negative, thallus is in the shape of a rod, and size is about 0.4-0.5 μ m 0.9-2.3 μ
M, single or arrangement in pairs.
It is measured through overtesting, L- pyrrolidinyl arylamine enzyme, the glutamy arylamine enzyme pNA, gamma-glutamyl of KDSPL-04 bacterial strain
Transferase, L-PROLINE arylamine enzyme, tyrosine arylamine enzyme, urease, ELLMAN reagent are positive；Lipase, phosphatase, ornithine are de-
Carboxylic acid, lysine depickling enzyme, histidine assimilation are negative；PEARLITOL 25C, D-Glucose, D-MANNOSE, D-Maltose, ancient sugar hair
Ferment is negative.
Step 3: through liquid chromatography for measuring in aseptic water, degradation capability of the bacterial strain to penicillin.
The content of thallus is 10 in every milliliter of culture solution in culture solution7A, the concentration of penicillin is 30mg/L, in 30 DEG C,
160rpm shaking table concussion degradation 6h, after measured, the degradation rate of penicillin is 100%.Meanwhile the bacterial strain and fermentation bacteria residue co-culture
One day, using liquid chromatography for measuring, which can reach 100% to the degradation rate of penicillin.It is demonstrated experimentally that degraded solutions body
System is tap water, KDSPL-04；For bacterial strain in pH 7.0-8.0,30 to 40 DEG C of well-growns of temperature can be using penicillin as sole carbon
Source growth, is one plant of very promising degradation bacteria.
Step 4: measurement KDSPL-02 bacterial strain 16S rDNA sequence, by the 16S rDNA sequence of the measurement with
The gene order that Genebank accession number is the 16S rDNA of D88006 carries out homology analysis.
Firstly, extracting the total DNA of KDSPL-04 bacterial strain using a small amount of extracting methods of DNA.Secondly, general using 16S rRNA
Primer 2 7F-1492R expands its 16S rRNA gene order, and (sequencing result is directly sequenced in the PCR product that amplification obtains
See SEQUENCE LISTING in sequence table), the 16SrRNA gene order of acquisition is inputted into GeneBank, passes through Blastn journey
All sequences in ordered pair database are compared analysis.
The 16SrRNA gene order of bacterial strain KDSPL-04 of the invention has higher with the existing type strain that variovorax belongs to
Similitude.Itself and Variovorax paradoxusLAM12373TSequence similarity be 99.42%.The bacterial strain belongs to
Variovorax sp. identifies that the bacterial strain is Variovorax sp., i.e. variovorax bacterium in combination with its physiological and biochemical property
Strain.
Embodiment 2: screening, identification and the degradation capability measurement to penicillin of variovorax bacterial strain
Step 1: taking 10g sludge to be contained in the 250mL conical flask of sterilizing, and the penicillin that 100mL concentration is 0.3g/L is added
In solution, in 30 DEG C of revolving speeds be 160r/min shaking table culture.After culture starts, switching in every 7 days is primary, with 10% (V/ when switching
V inoculum concentration) is inoculated into fresh penicillin solution domestication culture medium, and steps up the content of penicillin.Detect mould
The degradation rate of element, picks out best one group so as to used in isolated strains.
Step 2: by the bacteria suspension (1 × 10 of above-mentioned bacterial strains in 1mL7A/mL) it is linked into 100mL penicillin containing 30mg/L
Tap water in, take it is parallel three times, and with not inoculated bacteria but the tap water of the penicillin containing same concentrations is control.30℃,
160rpm shaking table culture is cultivated 2,4,6 hours respectively, and liquid chromatography detects the content of penicillin, and experimental result is shown in Table one.Drop
Solution 6 hours, selected bacterial strain can almost degrade penicillin.It is KDSPL-04 by obtained strain number.
One KDSPL-04 bacterial strain of table is in tap water to the degradation rate of penicillin
Step 3: measurement KDSPL-04 bacterial strain 16S rDNA sequence, by the 16S rDNA sequence of measurement with
The gene order that Genebank accession number is the 16S rDNA of D88006 carries out homology analysis, it is believed that KDSPL-04 bacterial strain belongs to
Variovorax sp. identifies that the bacterial strain is Variovorax sp., i.e. variovorax in combination with its physiological and biochemical property.
Embodiment 3: variovorax bacterial strain screening and the degradation capability measurement to penicillin in bacteria residue
Penicillin wet mycelia of giving up is crossed into 0.2mm sieve, the useless mycelia after weighing 10g sieving, and it is put into 100mL tap water
In, the variovorax thallus 6g (weight in wet base) obtained through having cultivated culture in 48 hours is added into the wet mycelia solution that gives up in 30 DEG C or so,
If 3 groups of parallel tests, to add penicillin mycelia but variovorax body is not added as control, acts on 0,2,4,6 hour and sampling respectively, efficiently
Liquid phase method detects the residual quantity of penicillin in mycelia.The result shows that variovorax thallus and penicillin mycelia under the conditions of 30 DEG C, make
After 6 hours, 100% can reach to the degradation rate of penicillin.
Degradation effect of the two KDSPL-04 bacterial strain of table to penicillin in bacteria residue
The beneficial effects of the present invention are:
1, the variovorax of degradable penicillin provided by the invention, the ability with efficient degradation penicillin, can be to hair
The penicillin in mycelium that ferment industry generates effectively is degraded, and to eliminate antibiotic residue in the useless mycelia of penicillin, can be used
The harmless treatment of fermentation bacteria residue, provides effective approach for the harmless treatment of mycelia in penicillin industrial production.
2, the variovorax of degradable penicillin provided by the invention can use penicillin as sole carbon source growth, be somebody's turn to do
Bacterial strain can reach 100% within 6 hours to the degradation capability of 1g/L penicillin.The bacterial strain can be used in penicillin industrial production
Ferment the harmless treatment of bacteria residue, the processing method is efficient, it is environmentally protective, economize on resources, processing cost it is low, completely eliminate blueness
Adverse effect of the useless mycelia of mycin to environment.
3, variovorax of the invention gene relevant to degradation and key enzyme are in biological prosthetic and purification and industry and biology
Field of medicaments has a wide range of applications.
Although the present invention is illustrated using above-mentioned preferred embodiment, the protection model that however, it is not to limit the invention
It encloses, anyone skilled in the art are not departing within the spirit and scope of the present invention, and opposite above-described embodiment carries out various changes
It is dynamic still to belong to the range that the present invention is protected with modification, therefore protection scope of the present invention subjects to the definition of the claims.
SEQUENCE LISTING
<110>Hebei University of Science and Technology
<120>a kind of variovorax of degradable penicillin
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1392
<212> DNA
<213> Variovorax sp.
<400> 1
acggcagcgc gggagcaatc ctggcggcga gtggcgaacg ggtgagtaat acatcggaac 60
gtgcccaatca gtgggggata acgcagcgaa agctgtgcta ataccgcata cgatctacgg 120
atgaaagcag gggatcgcaa gaccttgcgc gaatggagcg gccgatggca gattaggtag 180
ttggtgaggt aaaggctcac caagccttcg atctggagct ggtctgagag gacgaccagc 240
cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtggg gaattttgga 300
caatgggcgc aagcctgatc cagccatgcc gcgtgcagga tgaaggcctt cgggttgtaa 360
actgcttttg tacggaacga aacggccttt tctaataaag agggctaatg acggtaccgt 420
aagaataagc accggctaac tacgtgccag cagccgcggt aatacgtagg gtgcaagcgt 480
taatcggaat tactgggcgt aaagcgtgcg caggcggtaa tgtaagacag ttgtgaaatc 540
cccgggctca acctgggaac tgcatctgtg actgcattgc tggagtacgg cagaggggga 600
tggaattccg cgtgtagcag tgaaatgcgt agatatgcgg aggaacaccg atggcgaagg 660
caatcccctg ggcctgtact gacgctcatg cacgaaagcg tggggagcaa acaggattag 720
ataccctggt agtccacgcc ctaaacgatg tcaactggtt gttgggaatt cactttctca 780
gtaacgaagc taacgcgtga agttgaccgc ctggggagta cggccgcaag gttgaaactc 840
aaaggaattg acggggaccc gcacaagcgg tggatgatgt ggtttaattc gatgcaacgc 900
gaaaaacctt acccaccttt gacatgtacg gaattcgcca gagatggctt agtgctcgaa 960
agagaaccgt aacacaggtg ctgcatggct gtcgtcagct cgtgtcgtga gatgttgggt 1020
taagtcccgc aacgagcgca acccttgtca ttagttgcta cattcagttg ggcactctaa 1080
tgagactgcc ggtgacaaac cggaggaagg tggggatgac gtcaaagtcct catggccctt 1140
ataggtgggg ctacacacgt catacaatgg ctggtacaaa gggttgccaa cccgcgaggg 1200
ggagctaatc ccataaaacc agtcgtagtc cggatcgcag tctgcaactc gactgcgtga 1260
agtcggaatc gctagtaatc gtggatcaga atgtcacggt gaatacgttc ccgggtcttg 1320
tacacaccgc ccgtcacacc atgggagcgg gttctgccag aagtagttag cttaaccgca 1380
aggagggcga ta 1392
Claims (1)
1. a kind of variovorax of degradable penicillin (Variovorax sp.) KDSPL-04, which is characterized in that the variovorax with
Deposit number CGMCC No. 12495 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102787129A (en) * | 2012-07-20 | 2012-11-21 | 南京师范大学 | Amidase gene of Variovorax boronicumulans CGMCC 4969, and its application in biological degradation of acrylamide |
| CN105819442A (en) * | 2016-03-23 | 2016-08-03 | 河北科技大学 | Method for preparing activated carbon by means of denitrification of waste penicillin mycelia |
-
2016
- 2016-12-27 CN CN201611223442.8A patent/CN106591194B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102787129A (en) * | 2012-07-20 | 2012-11-21 | 南京师范大学 | Amidase gene of Variovorax boronicumulans CGMCC 4969, and its application in biological degradation of acrylamide |
| CN105819442A (en) * | 2016-03-23 | 2016-08-03 | 河北科技大学 | Method for preparing activated carbon by means of denitrification of waste penicillin mycelia |
Non-Patent Citations (2)
| Title |
|---|
| "茶园土壤抗生素抗性细菌及抗性基因研究";徐晨光;《中国优秀硕士学位论文全文数据库（电子期刊）农业科技辑》;20150115;第3.3.1-3.3.2节 * |
| 土壤中以抗生素为单一碳源的抗性细菌;顾超;《浙江大学学报（农业与生命科学版）》;20160918;第42卷(第5期);第582-588页 * |
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