基因组信息指导下茂源链霉菌代谢产物研究
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摘要
目的在基因组信息指导下,研究茂源链霉菌Streptomyces mobaraensis US-43的代谢产物。方法采用16S r RNA基因序列分析对菌株US-43进行初步分类鉴定,应用anti SMASH分析菌株US-43的基因组序列并预测其可能含有的代谢产物;采用正向硅胶柱色谱、凝胶柱色谱以及高效液相色谱等分离技术对该菌株的代谢产物进行分离纯化,利用核磁共振和质谱鉴定化合物结构;通过改变培养条件对其发酵稳定性进行考察,通过HPLC-UV方法建立标准曲线,研究代谢产物的产量。结果在基因组信息指导下从茂源链霉菌Streptomyces mobaraensis US-43中分离鉴定了一个α-吡啶酮类化合物杀粉蝶菌素A1,且该菌株的发酵稳定性良好,最高产量为144mg/L。结论茂源链霉菌Streptomyces mobaraensis US-43能够稳定产生杀粉蝶菌素A1。
Objective To research the metabolites of the strain Streptomyces mobaraensis US-43 guided by genome informatics. Methods Phylogenetic analyses of the strain US-43 were based on 16 S r RNA gene sequence. Antismash analysis was carried out to research the genome sequence of the strain US-43 and predict its potential metabolites. Then the target compound was isolated and purified with silica gel column chromatography, LH20 column chromatography, TLC and HPLC. The structure of the obtained compound was elucidated by NMR and mass spectrometry. The fermentation stability was investigated by changing the culture conditions, and the standard curve was established by the HPLC-UV method to study the yield of metabolites. Results Finally, an α-pyridone compound piericidin A1 was isolated and identified from the strain Streptomyces mobaraensis US-43 guided by genome informatics. The fermentation stability of the strain US-43 was good and its maximum yield was 144 mg/L. Conclusion The strain Streptomyces mobaraensis US-43 was able to stably produce piericidin A1.
Objective To research the metabolites of the strain Streptomyces mobaraensis US-43 guided by genome informatics. Methods Phylogenetic analyses of the strain US-43 were based on 16 S r RNA gene sequence. Antismash analysis was carried out to research the genome sequence of the strain US-43 and predict its potential metabolites. Then the target compound was isolated and purified with silica gel column chromatography, LH20 column chromatography, TLC and HPLC. The structure of the obtained compound was elucidated by NMR and mass spectrometry. The fermentation stability was investigated by changing the culture conditions, and the standard curve was established by the HPLC-UV method to study the yield of metabolites. Results Finally, an α-pyridone compound piericidin A1 was isolated and identified from the strain Streptomyces mobaraensis US-43 guided by genome informatics. The fermentation stability of the strain US-43 was good and its maximum yield was 144 mg/L. Conclusion The strain Streptomyces mobaraensis US-43 was able to stably produce piericidin A1.
引文
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[15]Iwasaki H,Kamisango K,Kuboniwa H,et al.3'-Deoxytalopiericidin-A1,a novel analog of antitumor antibiotics from oligotroph[J].J Antibiot,1991,44(4):451-452.
[16]Hall C,Wu M,Crane F L,et al.Piericidin A:A new inhibitor of mitochondrial electron transport[J].Biochem Biophys Res Commun,1966,25(4):373-377.
[17]Schnermann M J,Romero F A,Hwang I,et al.Total synthesis of piericidin A1 and B1 and key analogues[J].J Am Chem Soc,2006,128(36):11799-11807.
[18]Jeso V,Yang C,Cameron M D,et al.Synthesis and SAR of Lehualide B:A marine–derived natural product with potent anti–multiple myeloma activity[J].ACS Chem Biol,2013,8(6):1241-1252.
[19]Liu Q,Yao F,Chooi Y H,et al.Elucidation of piericidin A1 biosynthetic locus revealed a thioesterase-dependent mechanism ofα-pyridone ring formation[J].Chem Biol,2012,19(2):243-253.
[20]Singh B,Parshad R,Khajuria R K,et al.Saccharonol B,a new cytotoxic methylated isocoum arin from Saccharomonospora azurea[J].Tetrahed Lett,2013,54(49):6695-6699.
[21]Hwang J H,Kim J Y,Cha M R,et al.Etoposide-resistant HT-29 human colon carcinoma cells during glucose deprivation are sensitive to piericidin A,a GRP78 downregulator[J].J Cell Physiol,2008,215(1):243-250.
[2]Sidebottom A M,Carlson E E.A reinvigorated era of bacterial secondary metabolite discovery[J].Curr Opin Chem Biol,2015,24C:104-111.
[3]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.
[4]Zhou X F,Fenical W.The unique chemistry and biology of the piericidins[J].J Antibiot,2016,69(8):582-593.
[5]Hall C,Wu M,Crane F L,et al.Piericidin A:A new inhibitor of mitochondrial electron transport[J].Biochem Biophys Res Commun,1966,25(4):373-377.
[6]Takahashi N,Suzuki A,Kimura Y,et al.Structure of piericidin B and stereochemistry of piericidins[J]. Tetrahedron Lett,1967,21(21):1961-1964.
[7]Kubota N K,Ohta E,Ohta S,et al.Piericidins C5 and C6:New 4-pyridinol compounds produced by Streptomyces sp.and Nocardioides sp.[J].Bioorg Med Chem,2003,11(21):4569-4575.
[8]Hayakawa Y,Shirasaki S,Shiba S.Piericidins C7 and C8,new cytotoxic antibiotics produced by a marine Streptomyces sp.[J].J Antibiot,2007,60(3):196-200.
[9]Hayakawa Y,Shirasaki S,Kawasaki T,et al.Structures of new cytotoxic antibiotics,piericidins C7 and C8[J].J Antibiot,2007,60(35):201-203.
[10]Urakawa A,et al.IT-143-A and B,novel piericidin group antibiotics produced by Streptomyces sp.[J].J Antibiot,1996,49(10):1052-1055.
[11]Ueda J Y,Togashi T,Matukura S,et al.A novel nuclear export inhibitor JBIR-02,a new piericidin discovered from Streptomyces sp.ML55[J].J Antibiot,2007,60(7):459-462.
[12]Kimura K,Takahashi H,Miyata N,et al.New piericidin antibiotics,7-demethylpiericidin A1 and 7-demethyl-3'-rhamnopiericidin A1[J].J Antibiot,1996,27(49):697-699.
[13]Matsumoto M,Mogi K,Nagaoka K,et al.New piericidin glucosides,glucopiericidins A and B[J].J Antibiot,1987,40(2):149-156.
[14]Shaaban K A,Helmke E,Kelter G,et al.Glucopiericidin C:A cytotoxic piericidin glucoside antibiotic produced by a marine Streptomyces isolate[J].J Antibiot,2011,64(25):205-209.
[15]Iwasaki H,Kamisango K,Kuboniwa H,et al.3'-Deoxytalopiericidin-A1,a novel analog of antitumor antibiotics from oligotroph[J].J Antibiot,1991,44(4):451-452.
[16]Hall C,Wu M,Crane F L,et al.Piericidin A:A new inhibitor of mitochondrial electron transport[J].Biochem Biophys Res Commun,1966,25(4):373-377.
[17]Schnermann M J,Romero F A,Hwang I,et al.Total synthesis of piericidin A1 and B1 and key analogues[J].J Am Chem Soc,2006,128(36):11799-11807.
[18]Jeso V,Yang C,Cameron M D,et al.Synthesis and SAR of Lehualide B:A marine–derived natural product with potent anti–multiple myeloma activity[J].ACS Chem Biol,2013,8(6):1241-1252.
[19]Liu Q,Yao F,Chooi Y H,et al.Elucidation of piericidin A1 biosynthetic locus revealed a thioesterase-dependent mechanism ofα-pyridone ring formation[J].Chem Biol,2012,19(2):243-253.
[20]Singh B,Parshad R,Khajuria R K,et al.Saccharonol B,a new cytotoxic methylated isocoum arin from Saccharomonospora azurea[J].Tetrahed Lett,2013,54(49):6695-6699.
[21]Hwang J H,Kim J Y,Cha M R,et al.Etoposide-resistant HT-29 human colon carcinoma cells during glucose deprivation are sensitive to piericidin A,a GRP78 downregulator[J].J Cell Physiol,2008,215(1):243-250.