CN104789496A - Application of five myxobacteria to predation of drug-resistance bacteria and preparation of drug-resistance bacteria suppression drugs - Google Patents
Application of five myxobacteria to predation of drug-resistance bacteria and preparation of drug-resistance bacteria suppression drugs Download PDFInfo
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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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention discloses application of five myxobacteria to predation of drug-resistance bacteria and preparation of drug-resistance bacteria suppression drugs. The five myxobacteria, namely, Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus coralloides GIM1.816, are capable of preying drug-resistance bacteria and generating active natural products which are capable of remarkably suppressing the drug-resistance bacteria; therefore, the five myxobacteria have relatively good practical application value in the aspects of the biological control over the drug-resistance bacteria and the development of antibiotic drugs which are capable of effectively suppressing the drug-resistance bacteria.
Description
Technical field:
The invention belongs to microorganism field, be specifically related to five strain slime bacterias and suppress the application in resistant organism medicine at predation resistant organism with in preparation.
Background technology:
In the last few years, widely using and even abusing along with old microbiotic especially broad-spectrum antibiotics, various resistant organism was increasing and complicated.At present, the resistance of bacterium has become the significant threat of medical industry and human health.The new antibiotic kind of going on the market in recent years is less, and the means can taked these resistant organisms are very limited, and their appearance is almost a kind of disaster.Finding and developing efficiently to suppress the microbiotic of resistant organism very urgent.
Microbial natural products is the important sources of new antibiotic always.But as found streptomycete, subtilis, pseudomonas etc., new microbiotic is more and more difficult from traditional antibiotics generated bacterium.Meanwhile, the research finding new antibiotic producing strains is made slow progress again.In the last few years, cyanobacteria and this two quasi-microorganism of slime bacteria is only had to be developed to new antibiotics generated bacterium.
Slime bacteria is gram-negative soil original inhabitants bacterium, is also distributed widely in the various environment of occurring in nature simultaneously.Slime bacteria classification position is special, although belong to prokaryotic organism, but its a lot of feature is more similar to eukaryote, there is complicated social behavior and form occurs, as the formation etc. of multicellular signal conduction and induction, common coordinated movement, wolf pack formula predation, sporophore structure.
Slime bacteria is the important macromolecules degradation person of occurring in nature and microorganism predator, in the material cycle of microecological balance and global biosphere, play important role.Slime bacteria can produce the outer lyase of multiple born of the same parents, as lysis enzyme, nuclease, esterase, proteolytic enzyme, polysaccharidase, amylase and chitinase etc.The microorganisms such as the diversified bacterium of most of slime bacteria energy cracking, fungi, yeast and algae.Find that slime bacteria can the multiple pathogenic bacteria of cracking, there are important biological control potentiality.
Slime bacteria has genome maximum in prokaryotic organism, makes their secondary metabolite rich and varied.In slime bacteria, the ratio that can produce bioactive natural product is very high.In the slime bacteria of strain more than 2000 of molten bacterium, what can produce biologically active substance reaches 55%; In the slime bacteria of strain more than 700 of molten fiber, what can produce biologically active substance reaches 95%.The features such as the active substance that slime bacteria produces has novel structure, wide variety, active good, mechanism of action is complicated, at drug development, the aspect such as agriculture production and ecological management is widely used potentiality.Slime bacteria has become and has been only second to actinomycetic second largest antibiotics generated bacterium.In slime bacteria, find that at present more than 100 plant the secondary metabolite of brand new and the new structural derivative of kind more than 600, comprised heterocycle, aromatic nucleus, polyenoid, large ring, polyethers, alkaloid and peptide class.The antibacterium class medicine found in slime bacteria comprises Corallopyronin A, and Angiolam A, Thuggacins, Carolacton, Myxovirescins, Chondrochlorens etc. are multiple.
Summary of the invention:
The object of this invention is to provide five strain slime bacterias and suppress the application in resistant organism medicine at predation resistant organism with in preparation.
The present invention found through experiments, Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811, Corallococcusexiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus coralloides GIM1.816 has predation to resistance Salmonellas and antibiotic-resistance E. coli.Except Myxococcus sp.GIM1.815 to resistant Staphylococcus aureus without predation, other are as Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811, CorallococcusexiguusGIM1.813 and Corallococcus coralloides GIM1.816 has predation to resistant Staphylococcus aureus.
Therefore, first object of the present invention is to provide Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815 or the Corallococcus coralloidesGIM1.816 application in predation resistant organism.
Preferably, Myxococcus sp.GIM1.810, the application of Myxococcus sp.GIM1.811, CorallococcusexiguusGIM1.813 and Corallococcus coralloides GIM1.816 in predation resistance Salmonellas, antibiotic-resistance E. coli and resistant Staphylococcus aureus; The application of Myxococcus sp.GIM1.815 in predation resistance Salmonellas and antibiotic-resistance E. coli.
The present invention found through experiments:
Myxococcus sp.GIM1.810 can produce the medicine suppressing S.aureus 11, S.aureus 46, SalmonellaCMCC51005, Salmonella 56, E.coli A16 and E.coli D61-1.
Myxococcus sp.GIM1.811 can produce the medicine suppressing Staphylococcus aureus ATCC8739, S.aureus 15, S.aureus 46, Salmonella CMCC51005, Salmonella 31, Salmonella 56, Escherichia coli ATCC8739, E.coli A16, E.coli A29, E.coli D57 and E.coli D61-1.
Corallococcus exiguus GIM1.813 can produce the medicine suppressing Staphylococcus aureus ATCC8739, S.aureus11, S.aureus 15 and S.aureus 46.
Myxococcus sp.GIM1.815 can produce the medicine suppressing Salmonella CMCC51005, Salmonella 31, Salmonella47, Salmonella 56, Salmonella 59, Escherichia coli ATCC8739, E.coli A16, E.coli A29, E.coliD57 and E.coli D61-1.
Corallococcus coralloides GIM1.816 can produce the medicine suppressing Staphylococcus aureus ATCC8739, S.aureus 11, S.aureus 15, S.aureus 46.
Therefore, second object of the present invention is to provide Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815, Corallococcus coralloidesGIM1.816 are preparing the application in antibacterial medicines.
Further preferably, Myxococcus sp.GIM1.810 suppresses in preparation to apply in the medicine of S.aureus 11, S.aureus 46, SalmonellaCMCC51005, Salmonella 56, E.coli A16 and E.coli D61-1.
Myxococcus sp.GIM1.811 suppresses the application in the medicine of Staphylococcus aureus ATCC8739, S.aureus 15, S.aureus 46, Salmonella CMCC51005, Salmonella 31, Salmonella 56, intestinal bacteria or antibiotic-resistance E. coli in preparation.
Described antibiotic-resistance E. coli is E.coli A16, E.coli A29, E.coli D57 or E.coli D61-1.
Corallococcus exiguus GIM1.813 is preparing the application in the medicine suppressing streptococcus aureus or resistant Staphylococcus aureus.Described resistant Staphylococcus aureus is S.aureus 11, S.aureus 15 or S.aureus 46.
Myxococcus sp.GIM1.815 suppresses the application in the medicine of Salmonellas, resistance Salmonellas, intestinal bacteria or antibiotic-resistance E. coli in preparation.Described resistance Salmonellas is Salmonella 31, Salmonella 47, Salmonella 56, Salmonella 59, described antibiotic-resistance E. coli E.coli A16, E.coli A29, E.coli D57 and E.coli D61-1;
Corallococcus coralloides GIM1.816 is preparing the application in the medicine suppressing streptococcus aureus or resistant Staphylococcus aureus.Described resistant Staphylococcus aureus is S.aureus 11, S.aureus 15 or S.aureus 46.
Five strain slime bacteria Myxococcus sp.GIM1.810 of the present invention, Myxococcus sp.GIM1.811, Corallococcusexiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus coralloides GIM1.816 can prey on resistant organism, and it can also produce the bioactive natural product significantly suppressing resistant organism, therefore in the antibiotic medicine these resistant organisms being carried out to biological control or these resistant organisms of the effective suppression of exploitation, there is good actual application value.
Accompanying drawing illustrates:
Fig. 1 is the predation effects of five strain slime bacterias to resistant organism.
Embodiment:
Following examples further illustrate of the present invention, instead of limitation of the present invention.
Embodiment 1. is studied the predation of resistant organism by slime bacteria
The five strain slime bacterias of the present embodiment are respectively: Myxococcus sp.GIM1.810 (deposit number is GIM1.810), Myxococcus sp.GIM1.811 (deposit number is GIM1.811), Corallococcusexiguus GIM1.813 (deposit number is GIM1.813), Myxococcus sp.GIM1.815 (deposit number is GIM1.815) and Corallococcus coralloides GIM1.816 (deposit number is GIM1.816), above-mentioned five strain slime bacterias are all preserved in Guangdong Province's Culture Collection, its deposit number is shown in bacterial classification name bracket below, the bacterial strain at this preservation center is for sale, therefore anyone can buy this bacterial strain from this preservation center.
In the present embodiment, Resistant strain used is respectively: resistant Staphylococcus aureus three strain (S.aureus 11, S.aureus 15, S.aureus 46), Salmonellas four strain (Salmonella sp.31, Salmonella sp.47, Salmonella sp.56, Salmonellasp.59), intestinal bacteria four strain (E.coli A16, E.coli A29, E.coli D57, E.coli D61-1).There is provided by College of Veterinary Medicine, South China Agricultural University's pharmacology and Toxicology laboratory, concrete resistance information refers to table 1.
Table 1 resistant organism bacterial strain MIC value
Gram-negative bacteria
Note: μ g/L; MIC threshold value represents that being greater than threshold value is resistance to corresponding antibiotic strain; Intestinal bacteria and Salmonellas share a threshold value; Higher than threshold value bacterial strain, namely bacterial strain resists corresponding microbiotic, all indicates with overstriking.
1, above-mentioned five kinds of slime bacterias are inoculated respectively to CYE nutrient solution (10mM MOPS (3-(N-morpholine) propanesulfonic acid), 10g/L casein peptone, 5g/L yeast extract, 8mM MgSO
4, PH 7.6, solvent is water, and sterilization is for subsequent use) in, 150rpm, 30 DEG C of cultivation 3d, then use MMC damping fluid (10mM MOPS, 4mM MgSO
4, 2mM CaCl
2, PH 7.6, solvent is water, and sterilization is for subsequent use) rinse and be diluted to 1 × 10
11cell/ml.
2, inoculate Resistant strain in LB substratum, 150rpm, 37 DEG C of cultivations, to logarithmic phase, are then diluted to 1 × 10 with MMC wash buffer
9cell/mL.
3, by 20 μ L resistant organism bacterium drops to CFL solid medium (10mM MOPS, 1mM KH
2pO
4, 8mMMgSO
4, 0.2g/L (NH
4)
2sO
40.2g/L Trisodium Citrate, 0.2g/L Sodium.alpha.-ketopropionate, 0.1g/L junket peptone, 15g/L agar, pH 7.6, solvent is water, and sterilization is for subsequent use) on, after to be dried, its edge drip 1 μ L slime bacteria bacterium liquid (with prepared Chinese ink by volume 2:1 mix), two colony edges are at a distance of about 3mm.Flat board is placed in 32 DEG C of cultivations, 3,5,7, observe predation after 9d.
Concrete outcome as shown in Figure 1, as can be seen from Figure 1, the Myxococcus sp.GIM1.810 of the present embodiment, Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus coralloides GIM1.816 (illustrate only Salmonella sp.31 to resistance Salmonellas in figure, other resistance Salmonellass are identical results) and antibiotic-resistance E. coli (illustrate only E.coli A16 in figure, other antibiotic-resistance E. coli are identical results) all there is predation.Except Myxococcus sp.GIM1.815 to resistant Staphylococcus aureus without predation, other are as Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811, Corallococcus exiguusGIM1.813 and Corallococcus coralloides GIM1.816 has predation to resistant Staphylococcus aureus (illustrate only S.aureus 11 in figure, other resistant Staphylococcus aureus are identical results).
The meta-bolites of embodiment 2. slime bacteria is to the restraining effect research of resistant organism
1, at VY/2 substratum (fresh yeast 5.0g/L, CaCl
2 .2H
201.0g/L, VB120.0005g/L, PH 7.2) in inoculate five strain slime bacteria Myxococcus sp.GIM1.810 respectively, Myxococcus sp.GIM1.811, Corallococcusexiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus coralloidesGIM1.816,150rpm, cultivate 7d for 30 DEG C.
2,4000rpm is centrifugal, collects thalline and fermented liquid supernatant respectively.The isopyknic extraction into ethyl acetate 12h of fermented liquid.Ultrasonication after thalline soaks with acetone, is then extracted with ethyl acetate 12h.After extraction liquid rotary evaporation, use dissolve with methanol extract, fermented liquid and bacterial cell disruption liquid extract are dissolved into 50mg/mL and 100mg/mL concentration respectively.
3, inoculate resistant organism in LB liquid nutrient medium, 150rpm, cultivate logarithmic phase for 37 DEG C.Mix in the LB nutrient agar (liquid state) of 1:100 ratio with about 50 DEG C, shake up rear each plate and add 20mL.Flat board adheres to the 6mm diameter filter paper having dripped 5 μ L extraction liquids.Antibacterial circle diameter is measured after 37 DEG C of cultivation 18 ~ 24h.
4, five strain slime bacteria crude extracts to the inhibition of resistant organism in table 2.
The antibacterial circle diameter of table 2. five strain slime bacteria crude extract
B: fermentation broth coarse extract; C: thalline crude extract; Antibacterial circle diameter unit: mm;-, represent that crude extract does not have activity to corresponding resistant organism; In table, resistant organism used is with identical in embodiment 1, and wherein Staphylococcus aureus ATCC8739, Salmonella CMCC51005, Escherichia coli ATCC8739 are respectively streptococcus aureus, Salmonellas and colibacillary reference culture.
As can be seen from Table 2, Myxococcus sp.GIM1.810 can produce the medicine suppressing S.aureus 11, S.aureus 46, Salmonella CMCC51005, Salmonella 56, E.coli A16 and E.coli D61-1.
Myxococcus sp.GIM1.811 can produce the medicine suppressing Staphylococcus aureus ATCC8739, S.aureus 15, S.aureus 46, Salmonella CMCC51005, Salmonella 31, Salmonella 56, Escherichia coli ATCC8739, E.coli A16, E.coli A29, E.coli D57 and E.coli D61-1.
Corallococcus exiguus GIM1.813 can produce the medicine suppressing Staphylococcus aureus ATCC8739, S.aureus11, S.aureus 15 and S.aureus 46.
Myxococcus sp.GIM1.815 can produce the medicine suppressing Salmonella CMCC51005, Salmonella 31, Salmonella47, Salmonella 56, Salmonella 59, Escherichia coli ATCC8739, E.coli A16, E.coli A29, E.coliD57 and E.coli D61-1.
Corallococcus coralloides GIM1.816 can produce the medicine suppressing Staphylococcus aureus ATCC8739, S.aureus11, S.aureus 15, S.aureus 46.
Claims (10)
1.Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815 or the Corallococcus coralloides GIM1.816 application in predation resistant organism.
2. application according to claim 1, it is characterized in that, Myxococcus sp.GIM1.810, the application of Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813 and Corallococcus coralloides GIM1.816 in predation resistance Salmonellas, antibiotic-resistance E. coli and resistant Staphylococcus aureus; The application of Myxococcus sp.GIM1.815 in predation resistance Salmonellas and antibiotic-resistance E. coli.
3.Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815, Corallococcus coralloides GIM1.816 are preparing the application in antibacterial medicines.
4. application according to claim 3, it is characterized in that, Myxococcus sp.GIM1.810 suppresses the application in the medicine of S.aureus11, S.aureus 46, Salmonella CMCC51005, Salmonella 56, E.coli A16 or E.coli D61-1 in preparation.
5. application according to claim 3, it is characterized in that, Myxococcus sp.GIM1.811 suppresses the application in the medicine of Staphylococcus aureus ATCC8739, S.aureus 15, S.aureus 46, Salmonella CMCC51005, Salmonella 31, Salmonella 56, intestinal bacteria or antibiotic-resistance E. coli in preparation.
6. application according to claim 5, is characterized in that, described antibiotic-resistance E. coli is E.coli A16, E.coli A29, E.coli D57 or E.coli D61-1.
7. application according to claim 3, it is characterized in that, Corallococcus exiguus GIM1.813 or Corallococcus coralloides GIM1.816 is preparing the application in the medicine suppressing streptococcus aureus or resistant Staphylococcus aureus.
8. application according to claim 7, is characterized in that, described resistant Staphylococcus aureus is S.aureus 11, S.aureus 15 or S.aureus 46.
9. application according to claim 3, is characterized in that, Myxococcus sp.GIM1.815 suppresses the application in the medicine of Salmonellas, resistance Salmonellas, intestinal bacteria or antibiotic-resistance E. coli in preparation.
10. application according to claim 9, it is characterized in that, described resistance Salmonellas is Salmonella 31, Salmonella 47, Salmonella 56 or Salmonella 59, described antibiotic-resistance E. coli E.coli A16, E.coliA29, E.coli D57 or E.coli D61-1.
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