链霉菌Streptomyces sp. FJS31-2次级代谢产物UK-78623的分离鉴定及抑菌活性研究
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摘要
该研究对卤化II型聚酮类抗生素zunyimycins产生菌链霉菌(Streptomyces sp.)FJS31-2的次级代谢产物进行挖掘。次级代谢产物经乙酸乙酯萃取浓缩后,依次采用硅胶柱层析法、薄层层析(TLC)法、高效液相色谱(HPLC)法等方法进行分离和纯化,通过质谱(MS)法和核磁共振(NMR)等分析单体化合物的结构,并采用滤纸片法对其抗菌活性进行测试。结果表明,初次从该菌发酵物中分离得到多环内酯类化合物UK-78623,其对小肠结肠炎耶尔森氏菌(Yersinia enterocolitica)具有明显抑制作用,其抑菌圈直径为25.2 mm。
The secondary metabolites of Streptomyces sp. FJS31-2 producing halogenated type II polyketide antibiotics zunyimycins were further excavated. The secondary metabolite was extracted and concentrated by ethyl acetate, then isolated and purified by silica gel column chromatography,thin layer chromatography(TLC), HPLC and other methods. The structure of monomeric compound was analyzed by mass spectrometry and nuclear magnetic resonance method, and the antibacterial activity of the monomeric compound was determined by paper filtering method. The results showed that the polycyclic lactone compound UK-78623 was first isolated from the fermentation of Streptomyces sp. FJS31-2, which had a significant inhibiting effect on Yersinia enterocolitica, and the diameter of antibacterial circle was 25.2 mm.
The secondary metabolites of Streptomyces sp. FJS31-2 producing halogenated type II polyketide antibiotics zunyimycins were further excavated. The secondary metabolite was extracted and concentrated by ethyl acetate, then isolated and purified by silica gel column chromatography,thin layer chromatography(TLC), HPLC and other methods. The structure of monomeric compound was analyzed by mass spectrometry and nuclear magnetic resonance method, and the antibacterial activity of the monomeric compound was determined by paper filtering method. The results showed that the polycyclic lactone compound UK-78623 was first isolated from the fermentation of Streptomyces sp. FJS31-2, which had a significant inhibiting effect on Yersinia enterocolitica, and the diameter of antibacterial circle was 25.2 mm.
引文
[1] OCHOA J L, SANCHEZ L M, KOO B M, et al. Marine mammal microbiota yields novel antibiotic with potent activity against Clostridium difficile[J]. ACS Infect Dis, 2018, 4(1):59-67.
[2] KWON Y J, SOHN M J, KOSHINO H, et al. AN483, a new anti-MRSA compound from Streptomyces sp.[J]. J Antibiot, 2016, 69(10):762-764.
[3]王荫荫,王苗,钱声艳,等.链霉菌Streptomyces sp. FJS31-2产卤化二型聚酮类化合物的发酵条件优化[J].中国酿造,2017,36(1):66-69.
[4] LU Y H, SHAO M Y, WANG Y Y, et al. Zunyimycins B and C, New chloroanthrabenzoxocinones antibiotics against methicillin-resistant Staphylococcus aureus and Enterococci from Streptomyces sp. FJS31-2[J]. Molecules, 2017, 22(2):251.
[5] LU Y H, YUE C W, SHAO M Y, et al. Molecular genetic characterization of an anthrabenzoxocinones gene cluster in Streptomyces sp. FJS31-2 for the biosynthesis of BE-24566B and zunyimycin ale[J]. Molecules, 2016,21(6):711.
[6] BECERRIL A, ALVAREZ S, BRANA A F, et al. Uncovering production of specialized metabolites by Streptomyces argillaceus:Activation of cryptic biosynthesis gene clusters using nutritional and genetic approaches[J]. Plos one, 2018, 13(5):e0198145.
[7]江冰娅,赵薇,李书芬,等.链霉菌Streptomyces sp. CPCC 200497次级代谢产物Bohemamines的分离和鉴定[J].中国医药生物技术,2016,11(5):394-399.
[8]亢文佳,吴晟,华会明,等.基于基因组挖掘的一个新的吡嗪酮类天然产物的发现[J].有机化学,2016,36(7):1696-1699.
[9] KELLY C, CHALMERS J D, CROSSINGHAM I, et al. Macrolide antibiotics for bronchiectasis[J]. Cochrane Database Syst Rev, 2018, 3, Cd012406.
[10] GIACANI L, CICCATESE G, SALAZAR C P, et al. Enhanced molecular typing of treponema pallidum subspecies pallidum strains from 4 italian hospitals shows geographical differences in strain type heterogeneity,widespread resistance to macrolides, and lack of mutations associated with doxycycline resistance[J]. Sex Transmit Dis, 2018, 45(4):237-242.
[11] FOIK I P, TUSZYNSKA I, FEDER M, et al. Novel inhibitors of the r RNA ErmC'methyltransferase to block resistance to macrolides, lincosamides,streptogramine B antibiotics[J]. Eur J Med Chem, 2018, 146(2):60-67.
[12] PAWLOWSKI A C, STOGIOS P J, KOTEVA K, et al. The evolution of substrate discrimination in macrolide antibiotic resistance enzymes[J].Nat Commun, 2018, 9(1):112.
[13] THONG K L, TAN L K, OOI P T. Genetic diversity, virulotyping and antimicrobial resistance susceptibility of Yersinia enterocolitica isolated from pigs and porcine products in Malaysia[J]. J Sci Food Agr, 2018,98(1):87-95.
[14] HAXELL M A, BISHOP B F, BRYCE P, et al. C-13 beta-acyloxymilbemycins, a new family of macrolides. Discovery, structural determination and biological properties[J]. J Antibiot, 1992, 45(5):659-670.
[15] DUAN R, LIANG J, ZHANG J, et al. Prevalence of Yersinia enterocolitica bioserotype 3/O:3 among children with diarrhea, China, 2010-2015[J]. Emerg Infect Dis, 2017, 23(9):1502.
[16] LUCIANI M, SCHIRONE M, PORTANTI O, et al. Development of a rapid method for the detection of Yersinia enterocolitica serotype O:8from food[J]. Food Microbiol, 2018, 73(8):85-92.
[17] JUN J W, PARK S C, WICKLUND A, et al. Bacteriophages reduce Yersinia enterocolitica contamination of food and kitchenware[J]. Int J Food Microbiol, 2018, 271(8):33-47.
[18] FRAZAO M R, ANDRADE L N, DARINI A L, et al. Antimicrobial resistance and plasmid replicons in Yersinia enterocolitica strains isolated in Brazil in 30 years[J]. Brazil J Infect Dis, 2017, 21(4):477-480.
[19] LUNEC A. Superbugs and superdrugs-SMi's sixth annual conference. A focus on antibacterials[C]. Idrugs:the investigational drugs journal, 2004,7(4):323-326.
[20] JACKSON S A, CROSSMAN L, ALMEIDA E L, et al. Diverse and abundant secondary metabolism biosynthetic gene clusters in the genomes of marine sponge derived Streptomyces spp. isolates[J]. Marine Drugs,2018, 16(2):67.
[2] KWON Y J, SOHN M J, KOSHINO H, et al. AN483, a new anti-MRSA compound from Streptomyces sp.[J]. J Antibiot, 2016, 69(10):762-764.
[3]王荫荫,王苗,钱声艳,等.链霉菌Streptomyces sp. FJS31-2产卤化二型聚酮类化合物的发酵条件优化[J].中国酿造,2017,36(1):66-69.
[4] LU Y H, SHAO M Y, WANG Y Y, et al. Zunyimycins B and C, New chloroanthrabenzoxocinones antibiotics against methicillin-resistant Staphylococcus aureus and Enterococci from Streptomyces sp. FJS31-2[J]. Molecules, 2017, 22(2):251.
[5] LU Y H, YUE C W, SHAO M Y, et al. Molecular genetic characterization of an anthrabenzoxocinones gene cluster in Streptomyces sp. FJS31-2 for the biosynthesis of BE-24566B and zunyimycin ale[J]. Molecules, 2016,21(6):711.
[6] BECERRIL A, ALVAREZ S, BRANA A F, et al. Uncovering production of specialized metabolites by Streptomyces argillaceus:Activation of cryptic biosynthesis gene clusters using nutritional and genetic approaches[J]. Plos one, 2018, 13(5):e0198145.
[7]江冰娅,赵薇,李书芬,等.链霉菌Streptomyces sp. CPCC 200497次级代谢产物Bohemamines的分离和鉴定[J].中国医药生物技术,2016,11(5):394-399.
[8]亢文佳,吴晟,华会明,等.基于基因组挖掘的一个新的吡嗪酮类天然产物的发现[J].有机化学,2016,36(7):1696-1699.
[9] KELLY C, CHALMERS J D, CROSSINGHAM I, et al. Macrolide antibiotics for bronchiectasis[J]. Cochrane Database Syst Rev, 2018, 3, Cd012406.
[10] GIACANI L, CICCATESE G, SALAZAR C P, et al. Enhanced molecular typing of treponema pallidum subspecies pallidum strains from 4 italian hospitals shows geographical differences in strain type heterogeneity,widespread resistance to macrolides, and lack of mutations associated with doxycycline resistance[J]. Sex Transmit Dis, 2018, 45(4):237-242.
[11] FOIK I P, TUSZYNSKA I, FEDER M, et al. Novel inhibitors of the r RNA ErmC'methyltransferase to block resistance to macrolides, lincosamides,streptogramine B antibiotics[J]. Eur J Med Chem, 2018, 146(2):60-67.
[12] PAWLOWSKI A C, STOGIOS P J, KOTEVA K, et al. The evolution of substrate discrimination in macrolide antibiotic resistance enzymes[J].Nat Commun, 2018, 9(1):112.
[13] THONG K L, TAN L K, OOI P T. Genetic diversity, virulotyping and antimicrobial resistance susceptibility of Yersinia enterocolitica isolated from pigs and porcine products in Malaysia[J]. J Sci Food Agr, 2018,98(1):87-95.
[14] HAXELL M A, BISHOP B F, BRYCE P, et al. C-13 beta-acyloxymilbemycins, a new family of macrolides. Discovery, structural determination and biological properties[J]. J Antibiot, 1992, 45(5):659-670.
[15] DUAN R, LIANG J, ZHANG J, et al. Prevalence of Yersinia enterocolitica bioserotype 3/O:3 among children with diarrhea, China, 2010-2015[J]. Emerg Infect Dis, 2017, 23(9):1502.
[16] LUCIANI M, SCHIRONE M, PORTANTI O, et al. Development of a rapid method for the detection of Yersinia enterocolitica serotype O:8from food[J]. Food Microbiol, 2018, 73(8):85-92.
[17] JUN J W, PARK S C, WICKLUND A, et al. Bacteriophages reduce Yersinia enterocolitica contamination of food and kitchenware[J]. Int J Food Microbiol, 2018, 271(8):33-47.
[18] FRAZAO M R, ANDRADE L N, DARINI A L, et al. Antimicrobial resistance and plasmid replicons in Yersinia enterocolitica strains isolated in Brazil in 30 years[J]. Brazil J Infect Dis, 2017, 21(4):477-480.
[19] LUNEC A. Superbugs and superdrugs-SMi's sixth annual conference. A focus on antibacterials[C]. Idrugs:the investigational drugs journal, 2004,7(4):323-326.
[20] JACKSON S A, CROSSMAN L, ALMEIDA E L, et al. Diverse and abundant secondary metabolism biosynthetic gene clusters in the genomes of marine sponge derived Streptomyces spp. isolates[J]. Marine Drugs,2018, 16(2):67.