CN104789496B - Five plants of slime bacterias are in predation drug-fast bacteria and preparing the application in suppressing drug-fast bacteria medicine - Google Patents
Five plants of slime bacterias are in predation drug-fast bacteria and preparing the application in suppressing drug-fast bacteria medicine Download PDFInfo
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- 239000003814 drug Substances 0.000 title claims abstract description 29
- 241001357467 Myxococcus sp. Species 0.000 claims abstract description 45
- 241000588724 Escherichia coli Species 0.000 claims description 51
- 241000607142 Salmonella Species 0.000 claims description 44
- 241000191967 Staphylococcus aureus Species 0.000 claims description 19
- 230000001629 suppression Effects 0.000 claims description 5
- 241001478240 Coccus Species 0.000 claims description 2
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 2
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- 240000006365 Vitis vinifera Species 0.000 claims description 2
- 235000014787 Vitis vinifera Nutrition 0.000 claims description 2
- 241001647000 Corallococcus exiguus Species 0.000 abstract description 14
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- 230000000975 bioactive effect Effects 0.000 abstract description 4
- 230000002401 inhibitory effect Effects 0.000 abstract description 3
- 229930014626 natural product Natural products 0.000 abstract description 3
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- 241000191070 Escherichia coli ATCC 8739 Species 0.000 description 5
- 239000003242 anti bacterial agent Substances 0.000 description 5
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- GRCRWFPQJFBHLG-UHFFFAOYSA-N Angiolam A Natural products C=CC=C(C)C(O)CCC(C)C=CC1CCC(C)C(=O)CC(=O)NC(C)CCC(=O)C(C)C(O)C(C)C=C(C)C(=O)O1 GRCRWFPQJFBHLG-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000282461 Canis lupus Species 0.000 description 1
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- 241001646999 Corallococcus Species 0.000 description 1
- FPBHSTHTCPCNBS-UHFFFAOYSA-N Corallopyronin A Natural products COC(=O)NC=CCCC(C)C1=CC(=C(C(=O)C(=CC=C(/C)CCC(O)C(=C/CC=CC)C)C)C(=O)O1)O FPBHSTHTCPCNBS-UHFFFAOYSA-N 0.000 description 1
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- 238000012271 agricultural production Methods 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
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- WCKOGWVWLFJJJX-UHFFFAOYSA-N carolacton Natural products OC(=O)CC(OC)C(C)C(=O)C(C)C=C(C)C1OC(=O)C(O)C(O)C=CC(C)CCCC1C WCKOGWVWLFJJJX-UHFFFAOYSA-N 0.000 description 1
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- 238000006731 degradation reaction Methods 0.000 description 1
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical class CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 description 1
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- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
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- 238000002390 rotary evaporation Methods 0.000 description 1
- 230000009329 sexual behaviour Effects 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 235000011083 sodium citrates Nutrition 0.000 description 1
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical class [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
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- 231100000027 toxicology Toxicity 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical class [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
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Classifications
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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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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses five plants of slime bacterias in predation drug-fast bacteria and to prepare the application in suppressing drug-fast bacteria medicine.Five plants of slime bacteria Myxococcus sp.GIM1.810 of the present invention, Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus coralloides GIM1.816 can prey on drug-fast bacteria, and it can also produce the bioactive natural product for significantly inhibiting drug-fast bacteria, therefore have preferable actual application value in terms of these drug-fast bacterias are carried out with biological control or develops the antibiotic medicine for effectively suppressing these drug-fast bacterias.
Description
Technical field：
The invention belongs to microorganism field, and in particular to five plants of slime bacterias suppress drug-fast bacteria in predation drug-fast bacteria and preparing
Application in medicine.
Background technology：
In the last few years, widely using or even abusing with old antibiotic especially broad-spectrum antibiotic, various drug-fast bacterias
Increasing and complication.At present, the drug resistance of bacterium has turned into the significant threat of medical industry and human health.On in recent years
The antibiotics species in city is less, and the means that can be taken these drug-fast bacterias are very limited, and their appearance is almost a kind of disaster.
The antibiotic that searching and exploitation can efficiently suppress drug-fast bacteria is very urgent.
Microbial natural products are always the important sources of antibiotics.But from traditional antibiotics generated bacterium such as chain
Find that new antibiotic is more and more difficult in mould, bacillus subtilis, pseudomonad etc..Meanwhile find antibiotics and produce
The research of bacterium is made slow progress again.In the last few years, only this two quasi-microorganism of cyanobacteria and slime bacteria was developed to new antibiotic
Producing strains.
Slime bacteria is gram-negative soil original inhabitants bacterium, while is also distributed widely in the various environment in nature.It is viscous
Division bacteria status is special, although belonging to prokaryotes, its many feature and eucaryote are increasingly similar, has complicated society
Meeting sexual behaviour and form generation, such as multicellular signal conduction and sensing, the common coordinated movement of various economic factors, wolf pack formula predation, fructification
Formation of structure etc..
Slime bacteria is macromolecules degradation person and microorganism predator important in nature, is given birth in microecological balance and the earth
Important role is play in the material circulation of thing circle.Slime bacteria can produce a variety of extracellular lyases, such as cell lyases, core
Sour enzyme, esterase, protease, polysaccharase, amylase and chitinase etc..Most of slime bacterias can crack diversified bacterium,
The microorganisms such as fungi, yeast and algae.It has been found that slime bacteria can crack a variety of pathogens, there is important biological control to dive
Power.
Slime bacteria has genome maximum in prokaryotes so that their secondary metabolite is rich and varied.It is viscous thin
In bacterium, the ratio that can produce bioactive natural product is very high.In more than the 2000 plants of slime bacteria of molten bacterium, bioactive substance can be produced
Up to 55%；In more than 700 plants of slime bacteria of molten fiber, can produce bioactive substance up to 95%.It is active caused by slime bacteria
Material has the features such as structure novelty, wide variety, good, the mechanism of action complexity of activity, in drug development, agricultural production and life
State improvement etc. the potentiality that are widely used.Slime bacteria has become the second largest antibiotics generated bacterium for being only second to actinomyces.
It is found that the secondary metabolite of more than 100 kind brand news and the structural derivative that kind more than 600 is new, bag in slime bacteria at present
Include heterocycle, aromatic rings, polyenoid, big ring, polyethers, alkaloid and peptides.The antibacterium class medicine being had found in slime bacteria includes
Corallopyronin A,Angiolam A,Thuggacins,Carolacton,Myxovirescins,
Chondrochlorens etc. is a variety of.
The content of the invention：
It is an object of the invention to provide five plants of slime bacterias in predation drug-fast bacteria and answering in suppression drug-fast bacteria medicine is prepared
With.
The present invention is found through experiments that, Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811,
Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus
Coralloides GIM1.816 have predation to resistance salmonella and antibiotic-resistance E. coli.Except Myxococcus
Sp.GIM1.815 to resistant Staphylococcus aureus without predation, other such as Myxococcus sp.GIM1.810,
Myxococcus sp.GIM1.811, Corallococcusexiguus GIM1.813 and Corallococcus
Coralloides GIM1.816 have predation to resistant Staphylococcus aureus.
Therefore, first purpose of the invention is to provide Myxococcus sp.GIM1.810, Myxococcus
Sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815 or
Applications of the Corallococcus coralloides GIM1.816 in drug-fast bacteria is preyed on.
It is preferred that Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811,
Corallococcusexiguus GIM1.813 and Corallococcus coralloides GIM1.816 are husky in predation resistance
Application in door Salmonella, antibiotic-resistance E. coli and resistant Staphylococcus aureus；Myxococcus sp.GIM1.815 are being preyed on
Application in resistance salmonella and antibiotic-resistance E. coli.
The present invention is found through experiments that：
Myxococcus sp.GIM1.810, which can be produced, suppresses S.aureus 11, S.aureus 46, Salmonella
CMCC51005, Salmonella 56, E.coli A16 and E.coli D61-1 medicine.
Myxococcus sp.GIM1.811 can produce suppress 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 medicine
Thing.
Corallococcus exiguus GIM1.813, which can be produced, suppresses Staphylococcus aureus
ATCC8739, S.aureus11, S.aureus 15 and S.aureus 46 medicine.
Myxococcus sp.GIM1.815 can produce suppress Salmonella CMCC51005, Salmonella 31,
Salmonella47、Salmonella 56、Salmonella 59、Escherichia coli ATCC8739、E.coli
A16, E.coli A29, E.coli D57 and E.coli D61-1 medicine.
Corallococcus coralloides GIM1.816, which can be produced, suppresses Staphylococcus aureus
ATCC8739, S.aureus 11, S.aureus 15, S.aureus 46 medicine.
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,
Applications of the Corallococcus coralloides GIM1.816 in antibacterial medicines are prepared.
Further preferably, Myxococcus sp.GIM1.810 prepare suppress S.aureus 11, S.aureus 46,
Salmonella CMCC51005, Salmonella 56, E.coli A16 and E.coli D61-1 medicine in apply.
Myxococcus sp.GIM1.811 prepare suppress Staphylococcus aureus ATCC8739,
S.aureus 15, S.aureus 46, Salmonella CMCC51005, Salmonella 31, Salmonella 56, large intestine
Application in the medicine of bacillus or antibiotic-resistance E. coli.
Described antibiotic-resistance E. coli is E.coli A16, E.coli A29, E.coli D57 or E.coli D61-1.
Corallococcus exiguus GIM1.813 are preparing suppression staphylococcus aureus or resistant S Portugal
Application in the medicine of grape coccus.Described resistant Staphylococcus aureus be S.aureus 11, S.aureus 15 or
S.aureus 46。
Myxococcus sp.GIM1.815 are preparing suppression salmonella, resistance salmonella, Escherichia coli or resistance
Application in the medicine of Escherichia coli.Described resistance salmonella be 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 suppress staphylococcus aureus in preparation or resistance is golden yellow
Application in the staphylococcic medicine of color.Described resistant Staphylococcus aureus be S.aureus 11, S.aureus 15 or
S.aureus 46。
Five plants of slime bacteria Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811 of the present invention,
Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus
Coralloides GIM1.816 can prey on drug-fast bacteria, and it can also produce the natural production of activity for significantly inhibiting drug-fast bacteria
Thing, therefore have in terms of these drug-fast bacterias are carried out with biological control or develops the antibiotic medicine for effectively suppressing these drug-fast bacterias
There is preferable actual application value.
Brief description of the drawings：
Fig. 1 is predation effect of five plants of slime bacterias to drug-fast bacteria.
Embodiment：
Following examples are to further explanation of the invention, rather than limitation of the present invention.
The slime bacteria of embodiment 1. is studied the predation of drug-fast bacteria
Five plants of slime bacterias of the present embodiment are respectively：(deposit number is Myxococcus sp.GIM1.810
GIM1.810), My xococcus sp.GIM1.811 (deposit number GIM1.811), Corallococcusexiguus
GIM1.813 (deposit number GIM1.813), Myxococcus sp.GIM1.815 (deposit number GIM1.815) and
Corallococcus corallo ides GIM1.816 (deposit number GIM1.816), above-mentioned five plants of slime bacterias all preservations
In Guangdong Province's Culture Collection, its deposit number is shown in the bracket behind strain name, and the bacterial strain of the collection is
For sale, therefore anyone can buy the bacterial strain from the collection.
Antibody-resistant bacterium used is respectively in the present embodiment：Three plants of resistant Staphylococcus aureus (S.aureus 11,
S.aureus 15, S.aureus 46), four plants of salmonella (Salmonella sp.31, Salmonella sp.47,
Salmonella sp.56, Salmonella sp.59), Escherichia coli four plants of (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, specific resistance information
Refer to table 1.
The resistance bacteria strain MIC value of table 1
Gram-negative bacteria
Note：μg/L；It is the bacterial strain of resistance to corresponding antibiotic that MIC critical values, which are represented more than critical value,；Escherichia coli and Salmonella
Bacterium shares a critical value；Higher than critical value bacterial strain, the i.e. anti-corresponding antibiotic of bacterial strain, indicated with overstriking.
1st, be inoculated with respectively above-mentioned five kinds of slime bacterias to CYE nutrient solutions (10mM MOPS (3- (N- morpholines) propane sulfonic acid),
10g/L casein peptones, 5g/L yeast extracts, 8mM MgSO4, PH 7.6, solvent is water, and sterilization is standby) in,
150rpm, 30 DEG C of culture 3d, then with MMC buffer solutions (10mM MOPS, 4mM MgSO4, 2mM CaCl2, PH 7.6, solvent is
Water, sterilization are standby) rinse be diluted to 1 × 1011cell/ml。
2nd, antibody-resistant bacterium is inoculated with into LB culture mediums, and logarithmic phase is arrived in 150rpm, 37 DEG C of cultures, is then rushed with MMC buffer solutions
Wash and be diluted to 1 × 109cell/mL。
3rd, 20 μ L drug-fast bacteria bacterium solutions are dripped into CFL solid mediums (10mM MOPS, 1mM KH2PO4, 8mMMgSO4,
0.2g/L(NH4)2SO4, 0.2g/L sodium citrates, 0.2g/L Sodium Pyruvates, 0.1g/L junket peptones, 15g/L agar, pH 7.6, solvent
It is standby for water, sterilization) on, after to be dried, 1 μ L slime bacterias bacterium solution is added dropwise in its edge (with prepared Chinese ink by volume 2：1 is mixed
It is even), two colony edges are at a distance of about 3mm.Flat board is placed in 32 DEG C of cultures, 3,5,7, observe predation after 9d.
Concrete outcome as shown in figure 1, it will be seen from figure 1 that the Myxococcus sp.GIM1.810 of the present embodiment,
Myxococcus sp.GIM1.811, Corallococcus exiguus GIM1.813, Myxococcus sp.GIM1.815
With Corallococcus coralloides GIM1.816 (Salmonella illustrate only to resistance salmonella in figure
Sp.31, other resistance salmonellas are identical results) and antibiotic-resistance E. coli (E.coli A16 are illustrate only in figure, other
Antibiotic-resistance E. coli is identical result) all there is predation.Except Myxococcus sp.GIM1.815 are to resistant S
Staphylococcus without predation, other such as Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811,
Corallococcus exiguus GIM1.813 and Corallococcus coralloides GIM1.816 are golden yellow to resistance
Color staphylococcus (S.aureus 11 being illustrate only in figure, other resistant Staphylococcus aureus are identical results) all has
Predation.
Inhibitory action research of the metabolite of the slime bacteria of embodiment 2. to drug-fast bacteria
1st, in VY/2 culture mediums (yeast cake 5.0g/L, CaCl2 .2H201.0g/L, VB120.0005g/L, PH 7.2) in point
Not Jie Zhong five plants of slime bacteria Myxococcus sp.GIM1.810, Myxococcus sp.GIM1.811,
Corallococcusexiguus GIM1.813, Myxococcus sp.GIM1.815 and Corallococcus
GIM1.816,150rpm, 30 DEG C of culture 7d of coralloides.
2nd, 4000rpm is centrifuged, and collects thalline and fermented liquid supernatant respectively.Zymotic fluid is extracted with isometric ethyl acetate
12h.Ultrasonication after thalline acetone soak, is then extracted with ethyl acetate 12h.It is molten with methanol after extract rotary evaporation
Extract is solved, zymotic fluid and bacterial cell disruption liquid extract are dissolved into 50mg/mL and 100mg/mL concentration respectively.
3rd, drug-fast bacteria is inoculated with into LB fluid nutrient mediums, and logarithmic phase is arrived in 150rpm, 37 DEG C of cultures.By 1:100 ratios and 50
DEG C or so LB agar mediums (liquid condition) mixing, each plate adds 20mL after shaking up.Adhered on flat board and 5 μ L have been added dropwise
The 6mm diameter filter papers of extract.Antibacterial circle diameter is determined after 37 DEG C of 18~24h of culture.
4th, five plants of slime bacteria crude extracts are shown in Table 2 to the inhibition of drug-fast bacteria.
The antibacterial circle diameter of 2. 5 plants of slime bacteria crude extracts of table
B：Fermentation broth coarse extract；C：Thalline crude extract；Antibacterial circle diameter unit：mm；-, represent crude extract to corresponding resistance
Bacterium is without activity；Drug-fast bacteria used is identical with embodiment 1 in table, wherein Staphylococcus aureus ATCC8739,
Salmonella CMCC51005, Escherichia coli ATCC8739 be respectively staphylococcus aureus, salmonella and
The reference culture of Escherichia coli.
From table 2 it can be seen that Myxococcus sp.GIM1.810, which can be produced, suppresses S.aureus 11, S.aureus
46th, Salmonella CMCC51005, Salmonella 56, E.coli A16 and E.coli D61-1 medicine.
Myxococcus sp.GIM1.811 can produce suppress 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 medicine
Thing.
Corallococcus exiguus GIM1.813, which can be produced, suppresses Staphylococcus aureus
ATCC8739, S.aureus11, S.aureus 15 and S.aureus 46 medicine.
Myxococcus sp.GIM1.815 can produce suppress Salmonella CMCC51005, Salmonella 31,
Salmonella47、Salmonella 56、Salmonella 59、Escherichia coli ATCC8739、E.coli
A16, E.coli A29, E.coli D57 and E.coli D61-1 medicine.
Corallococcus coralloides GIM1.816, which can be produced, suppresses Staphylococcus aureus
ATCC8739, S.aureus11, S.aureus 15, S.aureus 46 medicine.
Claims (4)
1.Myxococcus sp.GIM1.810 are preparing the application in preying on drug-fast bacteria medicine.
2. application according to claim 1, it is characterised in that Myxococcus sp.GIM1.810 are preparing predation resistance
Application in salmonella, antibiotic-resistance E. coli and resistant Staphylococcus aureus medicine.
3.Myxococcus sp.GIM1.810 are preparing suppression resistance salmonella, antibiotic-resistance E. coli or resistant S Portugal
Application in grape coccus medicine.
4. application according to claim 3, it is characterised in that Myxococcus sp.GIM1.810 are preparing suppression
S.aureus 11, S.aureus 46, Salmonella CMCC51005, Salmonella 56, E.coli A16 or E.coli
Application in D61-1 medicine.
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