日照海岸线可培养放线菌的抑菌活性及其多样性研究
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摘要
本研究采用不同的筛选培养基及预处理方法对中国日照海岸线放线菌的分布、抗菌活性和多样性进行研究。结果表明,从中国日照海岸线样品中共分离到208株放线菌,其中91株放线菌具有抑菌作用,35株对产气杆菌有抑制作用,25株对变形杆菌有抑制作用,17株对金黄色葡萄球菌有抑制作用,9株对大肠杆菌有抑制作用,2株对铜绿假单胞菌有抑制作用,3株对黄曲霉有抑制作用。经分子生物学鉴定,25株抗菌活性较好的代表菌株可以分为22个种,3个不同的属:链霉菌属(Streptomyces sp.)、拟诺卡氏菌属(Nocardiopsis sp.)、糖单孢菌属(Saccharomonospora sp.),其中链霉菌属是优势菌属。
The distribution, antibacterial activity and diversity of Actinobacteria in the Rizhao coastline of China were studied by different screening medium and pretreatment methods. A total of 208 Actinobacteria were isolated from samples collected from the coastline. Among them, 91 Actinobacteria had antibacterial activity, 35 strains had antibacterial activities against Aerobacter aerogenes, 25 strains had antibacterial activities against Proteus, 17 strains had antibacterial activities against Staphylococcus aureus, 9 strains had antibacterial activities against Escherichia coli, 2 strains had antibacterial activities against Pseudomonas aeruginosa, and 3 strains had antibacterial activities against Aspergillus flavus. Through molecular biology identification, 25 representative strains with higher antibacterial activities were classified as 22 species belonging to 3 different genera: Streptomyces sp., Nocardiopsis sp. and Saccharomonospora sp., among them, Streptomyces sp. was the dominant genus.
The distribution, antibacterial activity and diversity of Actinobacteria in the Rizhao coastline of China were studied by different screening medium and pretreatment methods. A total of 208 Actinobacteria were isolated from samples collected from the coastline. Among them, 91 Actinobacteria had antibacterial activity, 35 strains had antibacterial activities against Aerobacter aerogenes, 25 strains had antibacterial activities against Proteus, 17 strains had antibacterial activities against Staphylococcus aureus, 9 strains had antibacterial activities against Escherichia coli, 2 strains had antibacterial activities against Pseudomonas aeruginosa, and 3 strains had antibacterial activities against Aspergillus flavus. Through molecular biology identification, 25 representative strains with higher antibacterial activities were classified as 22 species belonging to 3 different genera: Streptomyces sp., Nocardiopsis sp. and Saccharomonospora sp., among them, Streptomyces sp. was the dominant genus.
引文
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[23] ATALAN E, MANFIO G P, WARD A C, et al. Biosystematic studies on novel Streptomycetes from soil[J]. Anton Leeuw Int J G, 2000, 77(4):337-353.
[24]佘之蕴,黄宝莹,刘海卿,等.牛津杯法测定食品添加剂对五种益生菌的抑菌活力[J].食品工业,2016,37(1):171-174.
[25] TAMURA K, PETERSON D, PETERSON N, et al. MEGA5:Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Mol Biol Evol, 2011,28(10):2731-2739.
[26] SOLANO G, ROJAS-JIMéNEZ K, JASPARS M, et al. Study of the diversity of culturable Actinomycetes in the North Pacific and Caribbean coasts of Costa Rica[J]. Anton Leeuw Int J G, 2009, 96(1):71-78.
[27] WEBSTER N S, WILSON K J, BLACKALL L L, et al. Phylogenetic diversity of bacteria associated with the marine sponge Rhopaloeides odorabile[J]. Appl Environ Microbiol, 2001, 67(1):434-444.
[28]刘冬梅,李理,杨晓泉,等.用牛津杯法测定益生菌的抑菌活力[J].食品研究与开发,2006,27(3):145-147.
[29] PRIETO-DAVóA, TIAGO D, GOMES S E, et al. The Madeira Archipelago as a significant source of marine-derived Actinomycete diversity with anticancer and antimicrobial potential[J]. Front Microbiol, 2016, 7:1594.
[30] SATHISH K S R, RAO K V B. In-vitro antimicrobial activity of marine Actinobacteria against multidrug resistance Staphylococcus aureus[J].Asian Pac J Trop Med, 2012, 2(10):787-792.
[31] WATVE M G, TICKOO R, JOG M M, et al. How many antibiotics are produced by the genus Streptomyces?[J]. Arch Microbiol, 2001, 176(5):386-390.
[32] BENTLEY S D, CHATER K F, CERDENO-TARRAGA A M, et al.Complete genome sequence of the model Actinomycete Streptomyces coelicolor A3(2)[J]. Nature, 2002, 417(6885):141-147.
[33] BERDY J. Bioactive microbial metabolites[J]. J Antibiot, 2005, 58(1):1-26.
[34] DEUSHI T, YAMAGUCHI T, KAMIYA K, et al. A new aminoglycoside antibiotic, sannamycin C and its 4-N-glycyl derivative[J]. J Antibiot,1980, 33(11):1274-1280.
[35] ABDELMOHSEN U R, PIMENTEL-ELARDO S M, HANORA A, et al. Isolation, phylogenetic analysis and anti-infective activity screening of marine sponge-associated Actinomycetes[J]. Mar Drugs, 2010, 8(3):399-412.
[36] THIRUMALAIRAJ J, SIVASANKARI K, NATARAJASEENIVASAN K, et al. Potential anti-leptospiral compound, leptomycin B from marine Streptomyces indiaensis MSU5:taxonomy, fermentation, compound isolation, in vitro and in vivo efcacy[J]. World J Microbiol Biotechn,2017, 33(10):187.
[37] JOSEPH F, INIYAN A M, VINCENT S. HR-LC-MS based analysis of two antibacterial metabolites from a marine sponge symbiont Streptomyces pharmamarensis ICN40[J]. Microbial Pathogenesis, 2017, 111:450-457.
[38] LIMA S M A, MELO J G S, MILIT?O G C G, et al. Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon(Brazil)[J]. Appl microbiol Biot, 2017, 101(2):711-723.
[2] LAZZARINI A, CAVALETTI L, TOPPO G, et al. Rare genera of Actinomycetes as potential producers of new antibiotics[J]. Anton Leeuw Int J G, 2000, 78(3-4):399-405.
[3] BULL A T, WARD A C, GOODFELLOW M. Search and discovery strategies for biotechnology:the paradigm shift[J]. Microbiol Mol Biol R,2000, 64(3):573-606.
[4] BULL A T, STACH J E M. Marine Actinobacteria:new opportunities for natural product search and discovery[J]. Trends microbiol, 2007, 15(11):491-499.
[5] BUSARAKAM K, BROWN R, BULL A T, et al. Classification of thermophilic Actinobacteria isolated from arid desert soils, including the description of Amycolatopsis deserti sp. nov[J]. Anton Leeuw Int J G, 2016,109(2):319-334.
[6] LAHOUM A, VERHEECKE-VAESSEN C, BOURAS N, et al. Taxonomy of mycelial Actinobacteria isolated from Saharan soils and their efficiency to reduce aflatoxin B1content in a solid-based medium[J]. Ann Microbiol, 2017, 67(3):231-237.
[7] AZMAN A S, OTHMAN I, FANG C M, et al. Antibacterial, anticancer and neuroprotective activities of rare Actinobacteria from mangrove forest soils[J]. Indian J Microbiol, 2017, 57(2):177-187.
[8] DONIA M, HAMANN M T. Marine natural products and their potential applications as anti-infective agents[J]. Lancet Infect Dis, 2003, 3(6):338-348.
[9] ZHANG L, XI L, RUAN J, et al. Microbacterium marinum sp nov. isolated from deep-sea water[J]. Syst Appl Microbiol, 2012, 35(2):81-85.
[10] BECERRILESPINOSA A, FREEL K C, JENSEN P R, et al. Marine Actinobacteria from the Gulf of California:Diversity, abundance and secondary metabolite biosynthetic potential[J]. Anton Leeuw Int J G,2013, 103(4):809-819.
[11] FIEDLER H P, BRUNTNER C, BULL A T, et al. Marine Actinomycetes as a source of novel secondary metabolites[J]. Anton Leeuw Int J G, 2005, 87(1):37-42.
[12] LAM K S. Discovery of novel metabolites from marine Actinomycetes[J]. Curr Opin Microbiol, 2006, 9(3):245-251.
[13] RANJANI A, GOPINATH P M, RAJESH K, et al. Diversity of silver nanoparticle synthesizing Actinobacteria isolated from marine soil, Tamil Nadu[J]. Arabian J Sci Eng, 2016, 41(3):25-32.
[14] SARMIENTO-VIZCA?NO A, GONZáLEZ V, BRA?A A F, et al. Pharmacological potential of phylogenetically diverse Actinobacteria isolated from deep-sea coral ecosystems of the submarine Avilés Canyon in the Cantabrian sea[J]. Microbial Ecol, 2017, 73(2):338-352.
[15] MALDONADO L A, FRAGOSO-YA?EZ D, PéREZ-GARC?A A, et al.Actinobacterial diversity from marine sediments collected in Mexico[J].Anton Leeuw Int J G, 2009, 95(2):111-120.
[16] MAGARVEY N A, KELLER J M, BERNAN V, et al. Isolation and characterization of novel marine-derived Actinomycete taxa rich in bioactive metabolites[J]. Appl Environ Microbiol, 2004, 70(12):7520-7529.
[17] MALDONADO L A, FENICAL W, JENSEN PR, E T AL. Salinispora arenicola gen nov and Salinispora tropica sp. nov. obligate marine Actinomycetes belonging to the family Micromonosporaceae[J]. Int J Syst Evol Micr, 2005, 55(5):1759-1766.
[18] BULL A T, STACH J E M, WARD A C, et al. Marine Actinobacteria:perspectives, challenges, future directions[J]. Anton Leeuw Int J G,2005, 87(1):65-79.
[19] JENSEN P R, GONTANG E, MAFNAS C, et al. Culturable marine Actinomycete diversity from tropical Pacific Ocean sediments[J]. Environ Microbiol, 2005, 7(7):1039-1048.
[20] YUAN M, YU Y, LI R, et al. Phylogenetic diversity and biological activity of Actinobacteria isolated from the Chukchi Shelf marine sediments in the Arctic Ocean[J]. Mar Drugs, 2014, 12(3):1281-1297.
[21] PISANO A M, SOMMER J M, LOPEZ M M. Application of pretreatments for the isolation of bioactive Actinomycetes from marine sediments[J]. Appl Microbiol Biot, 1986, 25(3):285-288.
[22] TAKAHASHI Y, MATSUMOTO A, SEINO A, et al. Rare Actinomycetes isolated from desert soils[J]. Actinomycetologica, 1996, 10(2):91-97.
[23] ATALAN E, MANFIO G P, WARD A C, et al. Biosystematic studies on novel Streptomycetes from soil[J]. Anton Leeuw Int J G, 2000, 77(4):337-353.
[24]佘之蕴,黄宝莹,刘海卿,等.牛津杯法测定食品添加剂对五种益生菌的抑菌活力[J].食品工业,2016,37(1):171-174.
[25] TAMURA K, PETERSON D, PETERSON N, et al. MEGA5:Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Mol Biol Evol, 2011,28(10):2731-2739.
[26] SOLANO G, ROJAS-JIMéNEZ K, JASPARS M, et al. Study of the diversity of culturable Actinomycetes in the North Pacific and Caribbean coasts of Costa Rica[J]. Anton Leeuw Int J G, 2009, 96(1):71-78.
[27] WEBSTER N S, WILSON K J, BLACKALL L L, et al. Phylogenetic diversity of bacteria associated with the marine sponge Rhopaloeides odorabile[J]. Appl Environ Microbiol, 2001, 67(1):434-444.
[28]刘冬梅,李理,杨晓泉,等.用牛津杯法测定益生菌的抑菌活力[J].食品研究与开发,2006,27(3):145-147.
[29] PRIETO-DAVóA, TIAGO D, GOMES S E, et al. The Madeira Archipelago as a significant source of marine-derived Actinomycete diversity with anticancer and antimicrobial potential[J]. Front Microbiol, 2016, 7:1594.
[30] SATHISH K S R, RAO K V B. In-vitro antimicrobial activity of marine Actinobacteria against multidrug resistance Staphylococcus aureus[J].Asian Pac J Trop Med, 2012, 2(10):787-792.
[31] WATVE M G, TICKOO R, JOG M M, et al. How many antibiotics are produced by the genus Streptomyces?[J]. Arch Microbiol, 2001, 176(5):386-390.
[32] BENTLEY S D, CHATER K F, CERDENO-TARRAGA A M, et al.Complete genome sequence of the model Actinomycete Streptomyces coelicolor A3(2)[J]. Nature, 2002, 417(6885):141-147.
[33] BERDY J. Bioactive microbial metabolites[J]. J Antibiot, 2005, 58(1):1-26.
[34] DEUSHI T, YAMAGUCHI T, KAMIYA K, et al. A new aminoglycoside antibiotic, sannamycin C and its 4-N-glycyl derivative[J]. J Antibiot,1980, 33(11):1274-1280.
[35] ABDELMOHSEN U R, PIMENTEL-ELARDO S M, HANORA A, et al. Isolation, phylogenetic analysis and anti-infective activity screening of marine sponge-associated Actinomycetes[J]. Mar Drugs, 2010, 8(3):399-412.
[36] THIRUMALAIRAJ J, SIVASANKARI K, NATARAJASEENIVASAN K, et al. Potential anti-leptospiral compound, leptomycin B from marine Streptomyces indiaensis MSU5:taxonomy, fermentation, compound isolation, in vitro and in vivo efcacy[J]. World J Microbiol Biotechn,2017, 33(10):187.
[37] JOSEPH F, INIYAN A M, VINCENT S. HR-LC-MS based analysis of two antibacterial metabolites from a marine sponge symbiont Streptomyces pharmamarensis ICN40[J]. Microbial Pathogenesis, 2017, 111:450-457.
[38] LIMA S M A, MELO J G S, MILIT?O G C G, et al. Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon(Brazil)[J]. Appl microbiol Biot, 2017, 101(2):711-723.