Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Intra- and Extra-cellular Mechanisms of Saccharomyces cerevisiae Inactivation by High Voltage Pulsed Electric Fields Treatment

고전압 펄스 전기장에 의한 Saccharomyces cerevisiae의 세포내·외적 사멸 기작 연구

  • 이상재 (신라대학교 바이오식품소재학과) ;
  • 신정규 (전주대학교 한식조리학과)
  • Received : 2014.12.05
  • Accepted : 2014.12.24
  • Published : 2015.02.28
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Abstract

High voltage pulsed electric fields (PEF) treatment is one of the more promising nonthermal technologies to fully or partially replace thermal processing. The objective of this research was to investigate the microbial inactivation mechanisms of PEF treatment in terms of intra- and extracellular changes in the cells. Saccharomyces cerevisae cells treated with PEF showed cellular membrane damage. This resulted in the leakage of UV-absorbing materials and intracelluar ions, which increased with increasing treatment time and electric fields strength. This indicates that PEF treatment causes cell death via membrane damage and physical rupture of cell walls. We further confirmed this by Phloxine B staining, a dye that accumulates in dead cells. Using scanning and transmission electron microscopy, we observed morphological changes as well as disrupted cytoplasmic membranes in PEF treated S. cerevisae cells. In addition, PEF treatment led to damaged chromosomal DNA in S. cerevisiae.

비가열 살균 기술 중 본격적인 상업적 실용화를 눈앞에 두고 있는 고전압 펄스 전기장에 의한 미생물의 사멸 기작에 대해 살펴보았다. 세포 현탁액을 고전압 펄스 전기장 처리하였을 경우 처리 시간이나 전기장의 세기가 증가할수록 세포 외액으로 세포내 물질의 유출이 증가하였으며, 세포막 투과성의 변화로 인하여 $K^+$, $Na^+$등이 이온 성분의 유출도 나타났다. 염색시약에 의한 세포의 염색에서 처리시간이 증가함에 따라 염색되는 세포의 수가 증가하였으며, 전자현미경에 의한 세포의 관찰 결과 고전압 펄스 전기장 처리를 받은 세포의 경우 처리 받지 않은 것에 비해 표면이 거칠고 굴곡이 있었으며, 세포막이 터져 세포내 물질이 외부로 유출되고 형태가 일그러진 것이 관찰되었다. 항생물질 첨가에 따른 회복 실험에서 고전압 펄스 전기장 처리에 의해 세포의 단백질 합성 체계에 손상을 입었으며, chromosomal DNA의 분리를 통한 DNA의 손상여부 관찰 결과 약 27.3%의 DNA의 손상이 발생했음을 알 수 있었다. 따라서 고전압 펄스 전기장 처리가 세포벽이나 세포막의 손상뿐만 아니라 대사 체계와 DNA에도 손상을 주는 것을 확인할 수 있었다.

Keywords

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