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东方拟无枝酸菌中ECO-0501生物合成基因簇的筛选及其相关调控基因的初步分析 被引量：2

Screening ECO-0501 biosynthesis cluster from Amycolatopsis orientalis and preliminary analysis of related regulatory gene
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摘要 ECO-0501是从东方拟无枝酸菌代谢物中分离到的一种新型线性多烯类化合物,其不但具有良好的抗菌活性,且体内毒性较低,目前处于临床前的开发。本文通过对东方拟无枝酸菌的菌落原位杂交,筛选到4个粘粒克隆NL5B,NL9-3-1,NL3-1C,NL3-3A,可以覆盖整个ECO-0501生物合成簇,并对其中的基因ORF4进行了生物信息学分析,推测其为ECO-0501合成途径中的正调控基因。并通过该基因的过表达,成功地将ECO-0501的产量提高了6倍,为下一步ECO-0501的开发及其调控机制的研究打下基础。 ECO-0501 is a new linear polyene found from Amycolatopsis oriental＆. Because of the strong bacteridicial activity of ECO-0501 and its low toxity, preclinical development has been started on this antibiotic. Four positive cosmid clones were identified by colony hybridization that cover the whole ECO-0501 locus, including NL5B, NL9-3-1, NL3-1C and NL3-3A. Sequence analysis of the gene ORF4 revealed that its deduced protein might be a positive regulator controlling ECO-0501 biosynthesis. Overexpression of ORF4 showed that the yield of ECO- 0501 increased about six fold than that of wild-type strain. The present results provide us a tool for further study of ECO-0501 development and its regulatory mechanism.
作者 申洋 黄鹤 朱丽 杨晟 戈梅 陈代杰
机构地区 华东理工大学生物工程学院 中国科学院上海植物生理生态研究所 上海来益生物药物研究开发中心
出处 《中国抗生素杂志》 CAS CSCD 北大核心 2013年第5期327-331,共5页 Chinese Journal of Antibiotics
基金 国家自然科学基金(NFSC30672558) 国家重大新药创制项目(2012ZX090301002-003)
关键词 ECO-0501 东方拟无枝酸菌 菌落原位杂交 调控蛋白 ECO-0501 Amycolatopsis orientalis Colony hybridization Regulatory protein
分类号 Q784 [生物学—分子生物学] TQ465.7 [化学工程—制药化工]
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参考文献13

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共引文献9

同被引文献11

  • 1Banskota AH, Mcalpine JB, Sorensen D, et al. Genomic analyses lead to novel secondary metabolites. Part 3. ECO-0501, a novel antibacterial of a new class. J Antibiot (Tokyo), 2006, 59(9):533-542.
  • 2Alarco A, Dimitriadou V, McAlpine J, et al. ECO-0501, a novel anti-gram positive compound defining a new antibacterial structural class: preclinical evaluations. In: Proceedings of the 44th Annual ICAAC conference. Washington DC, 2004: Poster F-719. http://www. the freelibrary.com/Ecopia+Present s+Three+Posters+on+its+Antibacterial+Agent+ECO-0501+at...-a0123845408.
  • 3Shen Y, Huang H, Zhu L, et al. Type II thioesterase gene (ECO-orf27) from Amycolatopsis orientalis influences production of the polyketide antibiotic, ECO-0501 (LW01). Biotechnol Lett, 2012, 34(11):2087- 2091.
  • 4Shen Y, Huang H, Zhu L, et al. A regulatory gene (ECO-orf4) required for ECO-0501 biosynthesis in Amycolatopsis orientalis. J Basic Microbiol, 2014, 54(2): 104-110.
  • 5Stegmann E, Pelzer S, Wilken K, et al. Development of three different gene cloning systems for genetic investigation of the new species Amycolatopsis japonicum MG417-CF 17, the ethylacid producer. J Biotechnol, 2001, 92(2): 195-204.
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  • 7Rui Z, Petrickovi K, Skanta F, et al. Biochemical and genetic insights into asukamycin biosynthesis. J Biol Chem, 2010, 285(32):24915- 24924.
  • 8郦伟翔,阮林高,盛下放,戈梅,陈代杰.ECO-0501产生菌的发酵条件优化[J].中国医药工业杂志,2008,39(10):736-738. 被引量：4
  • 9Zhiqun Lu Pengfei Xie Zhongjun Qin.Promotion of markerless deletion of the actinorhodin biosynthetic gene cluster in Streptomyces coelicolor[J].Acta Biochimica et Biophysica Sinica,2010,42(10):717-721. 被引量：7
  • 10申洋,朱丽,阮林高,陈代杰.产N-demethyl ECO-0501工程菌的构建[J].中国抗生素杂志,2013,38(4):251-254. 被引量：1

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中国抗生素杂志
2013年 第5期

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