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Myxococcus xanthus의 protoporphyrin IX의 합성과 세포 성장에 대한 succinylacetone의 영향

The effects of succinylacetone on synthesis of protoporphyrin IX and cell growth of Myxococcus xanthus

  • 이병욱 (고신대학교 생명과학과)
  • 발행 : 2003.12.01

초록

Heme 합성의 중간체이며 또한 광수용체로도 작용하는 protoporphyrin IX의 세포 내 농도 및 성장 배지에 존재하는 농도가 야생형 M. xanthus DK1622 균주로부터 측정되었다. Protoporphyrin IX의 세포 내 농도는 배양 시간이 경과함에 따라 계속 증가하여, 안정기에 최고치에 이르는 것으로 나타났다 안정기에 도달한 세포 내에는 6.4 picomoles/mg of protein의 protoporphrin IX이 존재하는 것으로 밝혀졌다. Protoporphyrin IX은 대수기 중간 시기부터 세포외로 분비가 시작되어, 안정기에 도달한 세포의 배양액에서는 세포의 단백질 대비하여 3.0 picomoles/mg of protein이 존재하는 것으로 측정되었다. 영양분의 고갈에 기인하여 형성된 포자에서도 protoporphyrin IX의 농도는 6.5 picomoles/mg of protein이 존재하는 것으로 관찰되었다. Succinylacetone을 $500\muM$ 농도로 성장 배지에 첨가하였을 경우에 protoporphyrin IX의 생산은 검출이 불가능할 정도로 방해를 받았으며, 세포성장이 저해되고 세포 성장은 정상의 절반 수준인 약 100 Klett unit에서 정지하는 것으로 나타났다. 하지만 포자의 형성은 succinylacetone의 첨가에 관계없이 89-100%의 생성율을 보였음으로 정상 농도의 protoporphyrin IX가 M. xanthus의 성장을 위해서는 중요하지만, 포자 형성 과정에 필수적인 것으로 보이지는 않는다. 안정기 세포에서 나타나는 photolysis 현상도 succinylacetone의 첨가 여부에 관계없이 유사한 수준으로 관찰되었다.

Protoporphyrin IX is an intermediate molecule in the heme biosynthetic pathway. Intra- and extracellular concentrations of protoporphyrin IX in the wild type strain, Myxococcus xanthus DK1622 were measured by reverse phase HPLC. The amount of intracellular protoporphyrin IX continuously increased and reached 6.4 picomoles/mg of protein at the stationary phase. Extracellular protoporphyrin IX began to be detected from the mid-exponential phase. The culture supernatant that was collected in the stationary phase contained approximately 3.0 picomoles of proto-porphyrin IX per mg of protein. Spores formed by nutrient depletion contained about 6.5 picomole protoporphyrin IX/mg of protein. The synthesis of protoporphyrin IX and cell growth were strongly inhibited by addition of succinylacetone to a final concentration of $500\muM$. Succinylacetone, however did not appear to interfere developmental processes. Normal developmental behaviors including aggregation and spore formation was achieved even if succinylacetone was added in a medium. Photolysis among cells grown on a starvation medium supplemented with succinylacetone was also observed. These results indicate that protoporphyrin IX may be important to M. ,xanthus vegetative growth, but not critical to development processes.

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