• 中国中文核心期刊
  • 中国科学引文数据库核心期刊
  • 中国科技核心期刊
  • 中国高校百佳科技期刊
高级检索

庆大霉素生物合成基因genD2的研究

阙新桥, 陈泽斌, 洪文荣

阙新桥, 陈泽斌, 洪文荣. 庆大霉素生物合成基因genD2的研究[J]. 中国药科大学学报, 2014, 45(2): 237-241. DOI: 10.11665/j.issn.1000-5048.20140219
引用本文: 阙新桥, 陈泽斌, 洪文荣. 庆大霉素生物合成基因genD2的研究[J]. 中国药科大学学报, 2014, 45(2): 237-241. DOI: 10.11665/j.issn.1000-5048.20140219
QUE Xinqiao, CHEN Zebin, HONG Wenrong. Research on genD2 in gentamicin biosynthesis gene cluster[J]. Journal of China Pharmaceutical University, 2014, 45(2): 237-241. DOI: 10.11665/j.issn.1000-5048.20140219
Citation: QUE Xinqiao, CHEN Zebin, HONG Wenrong. Research on genD2 in gentamicin biosynthesis gene cluster[J]. Journal of China Pharmaceutical University, 2014, 45(2): 237-241. DOI: 10.11665/j.issn.1000-5048.20140219

庆大霉素生物合成基因genD2的研究

基金项目: 国家“重大新药创制”科技重大专项资助项目(No.2012ZX09201101-008);国家自然科学基金资助项目(No.31070093)

Research on genD2 in gentamicin biosynthesis gene cluster

  • 摘要: 以绛红小单孢菌G1008基因组为模板,PCR扩增genD2的上下游序列作为同源交换臂,在温敏型穿梭质粒pKC1139的基础上,构建同源重组质粒pDB303;通过接合转移,将质粒pDB303导入绛红小单孢菌G1008,经影印筛选得到一株genD2框内缺失的工程菌GD238。发酵并提取代谢产物,质谱分析表明,工程菌GD238只积累庆大霉素A2和A2e,证明genD2参与了庆大霉素加洛糖胺上C-3″位氨甲基化,可能是编码脱氢酶基因。
    Abstract: In order to verify the function of genD2, a recombinant plasmid pDB303 derivated from pKC1139 was constructed for blocking-up genD2. pDB303 was introduced into Micromonospora purpurea G1008 by conjugation. A desired mutant strain GD238 was obtained by PCR analysis. Antibiotics were isolated from the fermentation broth of GD238, the parent strain G1008 as the control. The result of mass spactral analysis indicated that the mutant strain GD238 only produce gentamicinA2 and A2e instead of gentamicinC complex, and that genD2 might be responsible for dehydrogenation at C-3″of garosamine.
  • [1] Kumar C,Himabindu M,Jetty A.Microbial biosynthesis and applications of gentamicin:a critical appraisal[J].Crit Rev Biotech,2008,28(3):173-212.
    [2] Yoshizawa S,Fourmy D,Puglisi JD.Structural origins of gentamicin antibiotic action[J].EMBO J,1998,17(22):6 437-6 448.
    [3] Dozzo P,Moser HE.New aminoglycoside antibiotics[J].Expert Opin Ther Pat,2010,20(10):1 321-1 341.
    [4] Endimiani A,Hujer KM,Hujer A M,et al.ACHN-490,a neoglycoside with potent in vitro activity against multidrug-resistant Klebsiella pneumoniae isolates[J].Antimicrob Agents Chemother,2009,53(10):4 504-4 507.
    [5] Nepal KK,Yoo JC,Sohng JK.Biosynthetic approach for the production of new aminoglycoside derivative[J].J Biosci Bioeng,2010,110(1):109-112.
    [6] Distler J,Braun C,Ebert A,et al.Gene cluster for streptomycin biosynthesis in Streptomyces griseus:analysis of a central region including the major resistance gene[J].Mol Gen Genet,1987,208(1/2):204-210.
    [7] Unwin J,Standage S,Alexander D,et al.Gene cluster in Micromonospora echinospora ATCC15835 for the biosynthesis of the gentamicin C complex[J].J.Antibiot,2004,57(7):436-445.
    [8] GenBank Database.icromonospora echinospora genomic region of the gentamicin biosynthesis gene cluster,strain DSM 43036[EB/OL].(2004-11-15),[2013-12-12] .http://www.ncbi.nlm.nih.gov/nuccore/85813995.
    [9] Kudo F,Eguchi T.Biosynthetic genes for aminoglycoside antibiotics[J].J Antibiot,2009,62(9):471-481.
    [10] Yan LB,Hong WR,Fang Z,et al.Construction of the conjugal transfer system for Micromonospora purpurea G1008[J].Chin J Antibiot(中国抗生素杂志),2011,12,36(12):899-904.
    [11] Hong W,Yan L.Identification of gntK,a gene required for the methylation of purpurosamine C-6′ in gentamicin biosynthesis[J].J Genapppl Microbiol,2011,58(5):349-356.
    [12] Kieser T,Bibb MJ,Mark J,et al.Practical Streptomyces Genetics[M].Norwich,England:The John Innes Foundation,2000:162-170.
    [13] Sambrook JF,Russell DW.Molecular Cloning:A Laboratory Manual[M].3rd ed.Cold Spring Harbor Laboratory Press,2001:27-99.
    [14] Chinese Pharmacopoeia Commission.Chinese Pharmacopoeia:part2(中华人民共和国药典:二部)[M].Beijing:China Medical Science Press,2010:附录 93-98.
    [15] Chen J,Xu L.Industrial analysis of antibiotics (抗生素工业分析)[M].Beijing:China Medical Science Press,1997:108-111.
    [16] Kim JY,Suh JW,Kang SH,et al.Gene inactivation study of gntE reveals its role in the first step of pseudotrisaccharide modifications in gentamicin biosynthesis[J].Biochem Biophys Res Commun,2008,372(4):730-734.
    [17] Park JW,Park SR,Nepal KK,et al.Discovery of parallel pathways of kanamycin biosynthesis allows antibiotic manipulation[J].Nat Chem Biol,2011,7(11):843-852.
    [18] Park SR,Park JW,Ban YH,et al.2-Deoxystreptamine-containing aminoglycoside antibiotics:recent advances in the characterization and manipulation of their biosynthetic pathways[J].Nat Prod Rep,2013,30(1):11-20.
计量
  • 文章访问数:  1624
  • HTML全文浏览量:  0
  • PDF下载量:  1960
  • 被引次数: 0
出版历程
  • 刊出日期:  2014-04-24

目录

    /

    返回文章
    返回
    x 关闭 永久关闭