CN107574130A - 珊瑚球菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 - Google Patents
珊瑚球菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 Download PDFInfo
- Publication number
- CN107574130A CN107574130A CN201710613426.8A CN201710613426A CN107574130A CN 107574130 A CN107574130 A CN 107574130A CN 201710613426 A CN201710613426 A CN 201710613426A CN 107574130 A CN107574130 A CN 107574130A
- Authority
- CN
- China
- Prior art keywords
- drug
- myxococcus
- coli
- aureus
- salmonella
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 241000894006 Bacteria Species 0.000 title claims abstract description 71
- 239000003814 drug Substances 0.000 title claims abstract description 28
- 241001478240 Coccus Species 0.000 title abstract description 5
- 235000014653 Carica parviflora Nutrition 0.000 title abstract description 4
- 244000132059 Carica parviflora Species 0.000 title description 3
- 241001085197 Corallococcus coralloides Species 0.000 claims abstract description 17
- 241000588724 Escherichia coli Species 0.000 claims description 48
- 241000607142 Salmonella Species 0.000 claims description 42
- 241000191967 Staphylococcus aureus Species 0.000 claims description 22
- 230000000844 anti-bacterial effect Effects 0.000 claims description 5
- 229940079593 drug Drugs 0.000 claims description 3
- 241001357467 Myxococcus sp. Species 0.000 abstract description 42
- 241001647000 Corallococcus exiguus Species 0.000 abstract description 14
- 230000003115 biocidal effect Effects 0.000 abstract description 9
- 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
- 241000243321 Cnidaria Species 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 14
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 241000191070 Escherichia coli ATCC 8739 Species 0.000 description 5
- 239000003242 anti bacterial agent Substances 0.000 description 5
- 229940088710 antibiotic agent Drugs 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- -1 polysaccharase Proteins 0.000 description 5
- 239000000287 crude extract Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 3
- 239000007993 MOPS buffer Substances 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical group C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 102000004317 Lyases Human genes 0.000 description 2
- 108090000856 Lyases Proteins 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 241000191940 Staphylococcus Species 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 229940066779 peptones Drugs 0.000 description 2
- 229930000044 secondary metabolite Natural products 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- GRCRWFPQJFBHLG-ZXXIUXGKSA-N (17e)-15-hydroxy-2-[(1e,7z)-6-hydroxy-3,7-dimethyldeca-1,7,9-trienyl]-5,10,14,16,18-pentamethyl-1-oxa-9-azacyclononadec-17-ene-6,8,13,19-tetrone Chemical compound C=C\C=C(\C)C(O)CCC(C)\C=C\C1CCC(C)C(=O)CC(=O)NC(C)CCC(=O)C(C)C(O)C(C)\C=C(C)\C(=O)O1 GRCRWFPQJFBHLG-ZXXIUXGKSA-N 0.000 description 1
- 241000186046 Actinomyces Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 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
- 102000012286 Chitinases Human genes 0.000 description 1
- 108010022172 Chitinases Proteins 0.000 description 1
- 229930189264 Chondrochloren Natural products 0.000 description 1
- 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
- 241000195493 Cryptophyta Species 0.000 description 1
- 241000192700 Cyanobacteria Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 0.000 description 1
- 229930189983 Myxovirescin Natural products 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 241000607149 Salmonella sp. Species 0.000 description 1
- 241000617098 Salmonella sp. 31 Species 0.000 description 1
- 241000807481 Salmonella sp. 56 Species 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 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
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 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
- WCKOGWVWLFJJJX-ZCXGUVEESA-N carolacton Chemical compound OC(=O)C[C@@H](OC)[C@@H](C)C(=O)[C@H](C)\C=C(/C)[C@H]1OC(=O)[C@H](O)[C@H](O)\C=C\[C@H](C)CCC[C@@H]1C WCKOGWVWLFJJJX-ZCXGUVEESA-N 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- FPBHSTHTCPCNBS-RAQHVQIZSA-N chembl1688851 Chemical compound COC(=O)N\C=C\CC[C@@H](C)C1=CC(O)=C(C(=O)C(\C)=C\C=C(/C)CCC(O)C(\C)=C\C\C=C\C)C(=O)O1 FPBHSTHTCPCNBS-RAQHVQIZSA-N 0.000 description 1
- 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
- 238000009509 drug development Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000000976 ink Substances 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 229920000570 polyether Polymers 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 244000062645 predators Species 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- KCXFHTAICRTXLI-UHFFFAOYSA-N propane-1-sulfonic acid Chemical compound CCCS(O)(=O)=O KCXFHTAICRTXLI-UHFFFAOYSA-N 0.000 description 1
- 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
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 229930187897 thuggacin Natural products 0.000 description 1
- 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
- 238000002525 ultrasonication Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
本发明公开了珊瑚球菌在捕食耐药菌和在制备抑制耐药菌药物中的应用。本发明的五株粘细菌Myxococcus sp.GIM1.810,Myxococcus sp.GIM1.811,Corallococcus exiguus GIM1.813,Myxococcus sp.GIM1.815和Corallococcus coralloides GIM1.816能够捕食耐药菌,并且其还能够产生显著抑制耐药菌的活性天然产物,因此在对这些耐药菌进行生物防治或者开发有效抑制这些耐药菌的抗生素药物方面具有较好的实际应用价值。
Description
本申请是专利申请号:201510152005.0,发明名称:五株粘细菌在捕食耐药菌和在制备抑制耐药菌药物中的应用,申请人:广东省微生物研究所的分案申请。
技术领域:
本发明属于微生物领域,具体涉及珊瑚球菌在捕食耐药菌和在制备抑制耐药菌药物中的应用。
背景技术:
近些年来,随着旧有抗生素尤其是广谱抗菌素的广泛使用乃至滥用,各种耐药菌日益增多和复杂化。目前,细菌的耐药性已成为医疗事业和人类健康的重大威胁。近些年上市的新抗生素种类较少,对这些耐药菌可采取的手段很有限,它们的出现几乎是一种灾难。寻找和开发能高效抑制耐药菌的抗生素刻不容缓。
微生物天然产物一直是新抗生素的重要来源。但是,从传统的抗生素产生菌如链霉菌、枯草芽孢杆菌、假单胞菌等中发现新的抗生素越来越困难。同时,寻找新抗生素产生菌的研究又进展缓慢。近些年来,只有蓝细菌和粘细菌这两类微生物被开发为新的抗生素产生菌。
粘细菌是革兰氏阴性的土壤土著菌,同时也广泛分布于自然界中的各种环境。粘细菌分类地位特殊,虽然属于原核生物,但其很多特征与真核生物更为相似,具有复杂的社会性行为和形态发生,如多细胞信号传导和感应、共同协调运动、狼群式捕食行为、子实体结构的形成等。
粘细菌是自然界中重要的大分子降解者和微生物捕食者,在微生态平衡和地球生物圈的物质循环中扮演着重要的角色。粘细菌能够产生多种胞外裂解酶,如细胞裂解酶、核酸酶、酯酶、蛋白酶、多糖酶、淀粉酶和几丁质酶等。大多数粘细菌能裂解多种多样的细菌、真菌、酵母和藻类等微生物。已发现粘细菌能够裂解多种病原菌,具有重要的生物防治潜力。
粘细菌具有原核生物中最大的基因组,使得它们的次级代谢产物丰富多样。粘细菌中,能产生活性天然产物的比例很高。溶细菌的2000多株粘细菌中,可产生生物活性物质的达55%;溶纤维的700多株粘细菌中,可产生生物活性物质的达95%。粘细菌产生的活性物质具有结构新颖、种类多样、活性良好、作用机制复杂等特征,在药物开发,农业生产和生态治理等方面有广泛的应用潜力。粘细菌已经成为仅次于放线菌的第二大抗生素产生菌。目前已在粘细菌中发现了100余种全新结构的次级代谢产物和600多种新的结构衍生物,包括杂环、芳香环、多烯、大环、聚醚、生物碱和肽类。粘细菌中已发现的抗细菌类药物包括Corallopyronin A,Angiolam A,Thuggacins,Carolacton,Myxovirescins,Chondrochlorens等多种。
发明内容:
本发明的目的是提供五株粘细菌在捕食耐药菌和在制备抑制耐药菌药物中的应用。
本发明通过实验发现,Myxococcus sp.GIM1.810,Myxococcus sp.GIM1.811,Corallococcus exiguus GIM1.813,Myxococcus sp.GIM1.815和Corallococcuscoralloides GIM1.816对耐药沙门氏菌和耐药大肠杆菌都具有捕食作用。除Myxococcussp.GIM1.815对耐药金黄色葡萄球菌无捕食作用,其他如Myxococcus sp.GIM1.810,Myxococcus sp.GIM1.811,Corallococcusexiguus GIM1.813和Corallococcuscoralloides GIM1.816对耐药金黄色葡萄球菌都具有捕食作用。
因此,本发明的第一个目的是提供Myxococcus sp.GIM1.810,Myxococcussp.GIM1.811,Corallococcus exiguus GIM1.813,Myxococcus sp.GIM1.815或Corallococcus coralloides GIM1.816在捕食耐药菌中的应用。
优选,Myxococcus sp.GIM1.810,Myxococcus sp.GIM1.811,Corallococcusexiguus GIM1.813和Corallococcus coralloides GIM1.816在捕食耐药沙门氏菌、耐药大肠杆菌和耐药金黄色葡萄球菌中的应用;Myxococcus sp.GIM1.815在捕食耐药沙门氏菌和耐药大肠杆菌中的应用。
本发明通过实验发现:
Myxococcus sp.GIM1.810能够产生抑制S.aureus 11、S.aureus 46、SalmonellaCMCC51005、Salmonella 56、E.coli A16和E.coli D61-1的药物。
Myxococcus sp.GIM1.811能产生抑制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和E.coli D61-1的药物。
Corallococcus exiguus GIM1.813能产生抑制Staphylococcus aureusATCC8739、S.aureus 11、S.aureus 15和S.aureus 46的药物。
Myxococcus sp.GIM1.815能产生抑制Salmonella CMCC51005、Salmonella 31、Salmonella 47、Salmonella 56、Salmonella 59、Escherichia coli ATCC8739、E.coliA16、E.coli A29、E.coli D57和E.coli D61-1的药物。
Corallococcus coralloides GIM1.816能产生抑制Staphylococcus aureusATCC8739、S.aureus 11、S.aureus 15、S.aureus 46的药物。
因此,本发明的第二个目的是提供Myxococcus sp.GIM1.810,Myxococcussp.GIM1.811,Corallococcus exiguus GIM1.813,Myxococcus sp.GIM1.815、Corallococcus coralloides GIM1.816在制备抑菌药物中的应用。
进一步优选,Myxococcus sp.GIM1.810在制备抑制S.aureus 11、S.aureus 46、Salmonella CMCC51005、Salmonella 56、E.coli A16和E.coli D61-1的药物中应用。
Myxococcus sp.GIM1.811在制备抑制Staphylococcus aureus ATCC8739、S.aureus 15、S.aureus 46、Salmonella CMCC51005、Salmonella 31、Salmonella 56、大肠杆菌或耐药大肠杆菌的药物中的应用。
所述的耐药大肠杆菌为E.coli A16、E.coli A29、E.coli D57或E.coli D61-1。
Corallococcus exiguus GIM1.813在制备抑制金黄色葡萄球菌或耐药金黄色葡萄球菌的药物中的应用。所述的耐药金黄色葡萄球菌为S.aureus 11、S.aureus 15或S.aureus 46。
Myxococcus sp.GIM1.815在制备抑制沙门氏菌、耐药沙门氏菌、大肠杆菌或耐药大肠杆菌的药物中的应用。所述的耐药沙门氏菌为Salmonella 31、Salmonella 47、Salmonella 56、Salmonella 59,所述的耐药大肠杆菌E.coli A16、E.coli A29、E.coliD57和E.coli D61-1;
Corallococcus coralloides GIM1.816在制备抑制金黄色葡萄球菌或耐药金黄色葡萄球菌的药物中的应用。所述的耐药金黄色葡萄球菌为S.aureus 11、S.aureus 15或S.aureus 46。
本发明的五株粘细菌Myxococcus sp.GIM1.810,Myxococcus sp.GIM1.811,Corallococcus exiguus GIM1.813,Myxococcus sp.GIM1.815和Corallococcuscoralloides GIM1.816能够捕食耐药菌,并且其还能够产生显著抑制耐药菌的活性天然产物,因此在对这些耐药菌进行生物防治或者开发有效抑制这些耐药菌的抗生素药物方面具有较好的实际应用价值。
附图说明:
图1是五株粘细菌对耐药菌的捕食效果。
具体实施方式:
以下实施例是对本发明的进一步说明,而不是对本发明的限制。
实施例1.粘细菌对耐药菌的捕食作用研究
本实施例的五株粘细菌分别为:Myxococcus sp.GIM1.810(保藏编号为GIM1.810),Myxococcus sp.GIM1.811(保藏编号为GIM1.811),CorallococcusexiguusGIM1.813(保藏编号为GIM1.813),Myxococcus sp.GIM1.815(保藏编号为GIM1.815)和Corallococcus coralloides GIM1.816(保藏编号为GIM1.816),上述五株粘细菌都保藏于广东省微生物菌种保藏中心,其保藏编号见菌种名后面的括号内,该保藏中心的菌株是对外销售的,因此任何人都可以从该保藏中心购买到该菌株。
本实施例中所用耐药菌株分别为:耐药金黄色葡萄球菌三株(S.aureus 11、S.aureus 15、S.aureus 46),沙门氏菌四株(Salmonella sp.31、Salmonella sp.47、Salmonella sp.56、Salmonellasp.59),大肠杆菌四株(E.coli A16、E.coli A29、E.coliD57、E.coli D61-1)。均由华南农业大学兽医学院药理与毒理学实验室提供,具体耐药信息详见表1。
表1耐药菌菌株MIC值
注:μg/L;MIC临界值表示大于临界值即为耐相应抗生素菌株;大肠杆菌及沙门氏菌共用一个临界值;高于临界值菌株,即菌株抗相应抗生素,均用加粗标示。
1、分别接种上述五种粘细菌到CYE培养液(10mM MOPS(3-(N-吗啡啉)丙磺酸),10g/L酪蛋白胨,5g/L酵母提取物,8mM MgSO4,PH 7.6,溶剂为水,灭菌消毒备用)中,150rpm、30℃培养3d,而后用MMC缓冲液(10mM MOPS,4mM MgSO4,2mM CaCl2,PH 7.6,溶剂为水,灭菌消毒备用)冲洗稀释至1×1011cell/ml。
2、接种耐药菌株到LB培养基中,150rpm、37℃培养到对数期,而后用MMC缓冲液冲洗稀释至1×109cell/mL。
3、将20μL耐药菌菌液滴到CFL固体培养基(10mM MOPS,1mM KH2PO4,8mM MgSO4,0.2g/L(NH4)2SO4,0.2g/L柠檬酸钠,0.2g/L丙酮酸钠,0.1g/L酪胨,15g/L琼脂,pH 7.6,溶剂为水,灭菌消毒备用)上,待干燥后,在其边缘处滴加1μL粘细菌菌液(与墨汁按体积比2:1混匀),两个菌落边缘相距约3mm。将平板置于32℃培养,3、5、7、9d后观察捕食现象。
具体结果如图1所示,从图1可以看出,本实施例的Myxococcus sp.GIM1.810,Myxococcus sp.GIM1.811,Corallococcus exiguus GIM1.813,Myxococcus sp.GIM1.815和Corallococcus coralloides GIM1.816对耐药沙门氏菌(图中只显示了Salmonellasp.31,其他耐药沙门氏菌是相同的结果)和耐药大肠杆菌(图中只显示了E.coli A16,其他耐药大肠杆菌是相同的结果)都具有捕食作用。除Myxococcus sp.GIM1.815对耐药金黄色葡萄球菌无捕食作用,其他如Myxococcus sp.GIM1.810,Myxococcus sp.GIM1.811,Corallococcus exiguus GIM1.813和Corallococcus coralloides GIM1.816对耐药金黄色葡萄球菌(图中只显示了S.aureus 11,其他耐药金黄色葡萄球菌是相同的结果)都具有捕食作用。
实施例2.粘细菌的代谢产物对耐药菌的抑制作用研究
1、在VY/2培养基(鲜酵母5.0g/L,CaCl2·2H20 1.0g/L,VB120.0005g/L,PH 7.2)中分别接种五株粘细菌Myxococcus sp.GIM1.810,Myxococcus sp.GIM1.811,Corallococcusexiguus GIM1.813,Myxococcus sp.GIM1.815和Corallococcuscoralloides GIM1.816,150rpm,30℃培养7d。
2、4000rpm离心,分别收集菌体和发酵液上清。发酵液用等体积的乙酸乙酯萃取12h。菌体用丙酮浸泡后超声破碎,然后用乙酸乙酯萃取12h。萃取液旋转蒸发后,用甲醇溶解萃取物,发酵液和菌体破碎液萃取物分别溶解成50mg/mL和100mg/mL浓度。
3、接种耐药菌到LB液体培养基中,150rpm,37℃培养到对数期。按1:100比例与50℃左右的LB琼脂培养基(液体状态)混合,摇匀后每个平皿加20mL。在平板上粘附已滴加5μL萃取液的6mm直径滤纸片。37℃培养18~24h后测定抑菌圈直径。
4、五株粘细菌粗提物对耐药菌的抑制效果见表2。
表2.五株粘细菌粗提物的抑菌圈直径
B:发酵液粗提物;C:菌体粗提物;抑菌圈直径单位:mm;—,表示粗提物对相应耐药菌没有活性;
表中所用耐药菌同实施例1中相同,其中Staphylococcus aureus ATCC8739、Salmonella CMCC51005、Escherichia coli ATCC8739分别为金黄色葡萄球菌、沙门氏菌和大肠杆菌的标准菌株。
从表2可以看出,Myxococcus sp.GIM1.810能够产生抑制S.aureus 11、S.aureus46、Salmonella CMCC51005、Salmonella 56、E.coli A16和E.coli D61-1的药物。
Myxococcus sp.GIM1.811能产生抑制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和E.coli D61-1的药物。
Corallococcus exiguus GIM1.813能产生抑制Staphylococcus aureusATCC8739、S.aureus11、S.aureus 15和S.aureus 46的药物。
Myxococcus sp.GIM1.815能产生抑制Salmonella CMCC51005、Salmonella 31、Salmonella47、Salmonella 56、Salmonella 59、Escherichia coli ATCC8739、E.coliA16、E.coli A29、E.coli D57和E.coli D61-1的药物。
Corallococcus coralloides GIM1.816能产生抑制Staphylococcus aureusATCC8739、S.aureus 11、S.aureus 15、S.aureus 46的药物。
Claims (5)
1.Corallococcus coralloides GIM1.816在捕食耐药菌中的应用。
2.根据权利要求1所述的应用,其特征在于,Corallococcus coralloides GIM1.816在捕食耐药沙门氏菌、耐药大肠杆菌和耐药金黄色葡萄球菌中的应用。
3.Corallococcus coralloides GIM1.816在制备抑菌药物中的应用。
4.根据权利要求3所述的应用,其特征在于,Corallococcus coralloides GIM1.816在制备抑制金黄色葡萄球菌或耐药金黄色葡萄球菌的药物中的应用。
5.根据权利要求4所述的应用,其特征在于,所述的耐药金黄色葡萄球菌为S.aureus11、S.aureus 15或S.aureus 46。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710613426.8A CN107574130B (zh) | 2015-04-01 | 2015-04-01 | 珊瑚球菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710613426.8A CN107574130B (zh) | 2015-04-01 | 2015-04-01 | 珊瑚球菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 |
CN201510152005.0A CN104789496B (zh) | 2015-04-01 | 2015-04-01 | 五株粘细菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201510152005.0A Division CN104789496B (zh) | 2015-04-01 | 2015-04-01 | 五株粘细菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN107574130A true CN107574130A (zh) | 2018-01-12 |
CN107574130B CN107574130B (zh) | 2020-07-28 |
Family
ID=53554691
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710613426.8A Active CN107574130B (zh) | 2015-04-01 | 2015-04-01 | 珊瑚球菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 |
CN201510152005.0A Active CN104789496B (zh) | 2015-04-01 | 2015-04-01 | 五株粘细菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201510152005.0A Active CN104789496B (zh) | 2015-04-01 | 2015-04-01 | 五株粘细菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (2) | CN107574130B (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113558069A (zh) * | 2021-09-26 | 2021-10-29 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | 一株捕食植物病原菌的黏细菌h56d21及其应用 |
CN116179400A (zh) * | 2022-08-17 | 2023-05-30 | 山东大学 | 一株产重排甾体化合物的黏细菌及其应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0064547B1 (de) * | 1980-11-19 | 1985-11-13 | Gesellschaft für Biotechnologische Forschung mbH (GBF) | Wirkstoffe und verfahren und mikroorganismen zu deren herstellung |
CN101638625A (zh) * | 2009-02-26 | 2010-02-03 | 淮阴工学院 | 橙色粘球菌jch-04及抗菌代谢产物的培养方法 |
CN102132788A (zh) * | 2011-03-22 | 2011-07-27 | 淮阴工学院 | 水产养殖用橙色粘球菌微生态制剂的制备及应用方法 |
CN103805542A (zh) * | 2014-01-23 | 2014-05-21 | 广东省微生物研究所 | 一种用于粘细菌菌体规模制备的液体发酵方法 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101659929B (zh) * | 2009-09-11 | 2010-09-22 | 中国农业大学 | 用于降解黄曲霉毒素b1的粘球菌菌株及其活性蛋白 |
-
2015
- 2015-04-01 CN CN201710613426.8A patent/CN107574130B/zh active Active
- 2015-04-01 CN CN201510152005.0A patent/CN104789496B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0064547B1 (de) * | 1980-11-19 | 1985-11-13 | Gesellschaft für Biotechnologische Forschung mbH (GBF) | Wirkstoffe und verfahren und mikroorganismen zu deren herstellung |
CN101638625A (zh) * | 2009-02-26 | 2010-02-03 | 淮阴工学院 | 橙色粘球菌jch-04及抗菌代谢产物的培养方法 |
CN102132788A (zh) * | 2011-03-22 | 2011-07-27 | 淮阴工学院 | 水产养殖用橙色粘球菌微生态制剂的制备及应用方法 |
CN103805542A (zh) * | 2014-01-23 | 2014-05-21 | 广东省微生物研究所 | 一种用于粘细菌菌体规模制备的液体发酵方法 |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113558069A (zh) * | 2021-09-26 | 2021-10-29 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | 一株捕食植物病原菌的黏细菌h56d21及其应用 |
CN116179400A (zh) * | 2022-08-17 | 2023-05-30 | 山东大学 | 一株产重排甾体化合物的黏细菌及其应用 |
CN116179400B (zh) * | 2022-08-17 | 2024-03-26 | 山东大学 | 一株产重排甾体化合物的黏细菌及其应用 |
Also Published As
Publication number | Publication date |
---|---|
CN107574130B (zh) | 2020-07-28 |
CN104789496A (zh) | 2015-07-22 |
CN104789496B (zh) | 2018-04-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Seema et al. | In vitro evaluation of biological control agents against Rhizoctonia solani. | |
CN107245457A (zh) | 一种铁皮石斛内生真菌菌株及其产生的胞外多糖以及该胞外多糖的提取方法与应用 | |
CN104195067B (zh) | 一株解淀粉芽孢杆菌及其在水产养殖中的应用 | |
Bartolini et al. | Stress-responsive alternative sigma factor SigB plays a positive role in the antifungal proficiency of Bacillus subtilis | |
CN107151641A (zh) | 一株抑制水稻纹枯病菌的侧孢短芽孢杆菌及其应用 | |
CN113142245B (zh) | 球孢白僵菌sb038和乙基多杀菌素对普通大蓟马的协同防治 | |
Huang et al. | The identification of a bacterial strain BGI-1 isolated from the intestinal flora of Blattella germanica, and its anti-entomopathogenic fungi activity | |
CN106967631B (zh) | 粘细菌在制备捕食和抑制植物病原细菌的药物中的应用 | |
CN107142226A (zh) | 一种枯草芽胞杆菌atr2及制备方法和应用 | |
CN104498443B (zh) | 鲍曼不动杆菌噬菌体及其应用 | |
KR101922410B1 (ko) | 바실러스 오리지콜라 yc7011이 생산하는 식물 병해충 방제 효과를 나타내는 기주 저항성 유도 신규 물질 | |
CN104789496B (zh) | 五株粘细菌在捕食耐药菌和在制备抑制耐药菌药物中的应用 | |
CN103232940B (zh) | 淡色生赤壳菌Bo-1菌株及其培养物和在拮抗病原细菌中的应用 | |
Chen et al. | Antifungal activity of Streptomyces albidoflavus L131 against the leaf mold pathogen Passalora fulva involves membrane leakage and oxidative damage | |
KR101175532B1 (ko) | 신규한 버크홀더리아 균주, 버크홀더리아 균주의 항진균성을 증가시키는 효과가 있는 미생물 대량생산용 배지 및 이 배지를 이용한 대량생산방법 | |
KR20100017997A (ko) | 식물병원균에 대한 항균활성을 갖는 스트렙토마이세스 라벤둘래 cmc0992[kctc18169p] 및 이를 이용한 식물병원균 방제제 | |
CN105018395B (zh) | 一株短小芽孢杆菌及其在防治苹果斑点落叶病中的应用 | |
CN109699683B (zh) | 一种滑石基质爪哇虫草孢子制剂 | |
Lin et al. | Assembly of an active microbial consortium by engineering compatible combinations containing foreign and native biocontrol bacteria of kiwifruit | |
CN108715817A (zh) | 一种微白黄链霉菌菌株及其应用 | |
Abdullah et al. | Improving the efficiency of Bacillus thuringiensis against insects of different feeding habits by plasmid transfer technique | |
CN103222483B (zh) | 盐屋链霉菌代谢产物在防治番茄灰霉病菌中的应用 | |
CN103222481B (zh) | 盐屋链霉菌代谢产物在防治水稻纹枯病菌中的应用 | |
CN114009451B (zh) | 深海放线菌11791在制备防治农牧业鳞翅目害虫药物中的应用 | |
CN103222478B (zh) | 盐屋链霉菌代谢产物在防治西瓜枯萎病菌中的应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CP03 | Change of name, title or address | ||
CP03 | Change of name, title or address |
Address after: No. 56, courtyard, No. 100, Xianlie Middle Road, Guangzhou, Guangdong 510070 Patentee after: Institute of Microbiology, Guangdong Academy of Sciences Country or region after: China Address before: No. 56, courtyard, No. 100, Xianlie Middle Road, Guangzhou, Guangdong 510070 Patentee before: GUANGDONG INSTITUTE OF MICROBIOLOGY (GUANGDONG DETECTION CENTER OF MICROBIOLOGY) Country or region before: China |