Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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In Vivo Characterization of Phosphotransferase-Encoding Genes istP and forP as Interchangeable Launchers of the C3',4'-Dideoxygenation Biosynthetic Pathway of 1,4-Diaminocyclitol Antibiotics

  • Nguyen, Lan Huong (Department of Biotechnology Convergent Pharmaceutical Engineering, SunMoon University) ;
  • Lee, Na Joon (Department of Integrated Biomedical and Life Sciences, Graduate School, Korea University) ;
  • Hwang, Hyun Ha (Department of Integrated Biomedical and Life Sciences, Graduate School, Korea University) ;
  • Son, Hye Bin (Department of Integrated Biomedical and Life Sciences, Graduate School, Korea University) ;
  • Kim, Hye Ji (Department of Integrated Biomedical and Life Sciences, Graduate School, Korea University) ;
  • Seo, Eun Gyo (Department of Integrated Biomedical and Life Sciences, Graduate School, Korea University) ;
  • Nguyen, Huu Hoang (Department of Biotechnology Convergent Pharmaceutical Engineering, SunMoon University) ;
  • Park, Je Won (Department of Integrated Biomedical and Life Sciences, Graduate School, Korea University)
  • Received : 2018.09.02
  • Accepted : 2019.01.07
  • Published : 2019.03.28
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Abstract

Deactivation of aminoglycosides by their modifying enzymes, including a number of aminoglycoside O-phosphotransferases, is the most ubiquitous resistance mechanism in aminoglycoside-resistant pathogens. Nonetheless, in a couple of biosynthetic pathways for gentamicins, fortimicins, and istamycins, phosphorylation of aminoglycosides seems to be a unique and initial step for the creation of a natural defensive structural feature such as a 3',4'-dideoxy scaffold. Our aim was to elucidate the biochemical details on the beginning of these C3',4'-dideoxygenation biosynthetic steps for aminoglycosides. The biosynthesis of istamycins must surely involve these 3',4'-didehydroxylation steps, but much less has been reported in terms of characterization of istamycin biosynthetic genes, especially about the phosphotransferase-encoding gene. In the disruption and complementation experiments pointing to a putative gene, istP, in the genome of wild-type Streptomyces tenjimariensis, the function of the istP gene was proved here to be a phosphotransferase. Next, an in-frame deletion of a known phosphotransferase-encoding gene forP from the genome of wild-type Micromonospora olivasterospora resulted in the appearance of a hitherto unidentified fortimicin shunt product, namely 3-O-methyl-FOR-KK1, whereas complementation of forP restored the natural fortimicin metabolite profiles. The bilateral complementation of an istP gene (or forP) in the ${\Delta}forP$ mutant (or ${\Delta}istP$ mutant strain) successfully restored the biosynthesis of 3',4'-dideoxy fortimicins and istamycins, thus clearly indicating that they are interchangeable launchers of the biosynthesis of 3',4'-dideoxy types of 1,4-diaminocyclitol antibiotics.

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