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Immuno-Enhancing Effect of Enzymatic Extract of Sargassum coreanum Using Crude Enzyme from Shewanella oneidensis PKA 1008

Shewanella oneidensis PKA 1008 유래 조효소 처리에 의한 큰잎모자반(Sargassum coreanum) 추출 분해물의 면역증진 효과

  • Park, Sun-Hee (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Kim, Min-Ji (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Kim, Go-Eun (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Park, So-Yeong (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Kim, Koth-Bong-Woo-Ri (Department of Food Science & Technology/Institute of Food Science, Pukyong National University) ;
  • Kim, Yeon-Ji (Institute of Fisheries Sciences, Pukyong National University) ;
  • Cho, Young-Je (School of Food Science of Biotechnology, Kyungpook National University) ;
  • Ahn, Dong-Hyun (Department of Food Science & Technology/Institute of Food Science, Pukyong National University)
  • 박선희 (부경대학교 식품공학과/식품연구소) ;
  • 김민지 (부경대학교 식품공학과/식품연구소) ;
  • 김고은 (부경대학교 식품공학과/식품연구소) ;
  • 박소영 (부경대학교 식품공학과/식품연구소) ;
  • 김꽃봉우리 (부경대학교 식품공학과/식품연구소) ;
  • 김연지 (부경대학교 수산과학연구소) ;
  • 조영제 (경북대학교 식품공학부) ;
  • 안동현 (부경대학교 식품공학과/식품연구소)
  • Received : 2017.05.22
  • Accepted : 2017.07.18
  • Published : 2017.08.31

Abstract

The immuno-enhancing effects of alginate oligosaccharides from Sargassum coreanum were investigated. The alginate oligosaccharides were produced by an alginate-degrading enzyme from S. oneidensis PKA 1008. The degraded alginate oligosaccharides were visualized by thin-layer chromatography developed using a solvent system of 1-butanol/methanol/water, 4:1:2 (v/v/v). Alginate was degraded into dimmers at 60 h. As a result, the levels of Th1 cytokine [interferon $(IFN)-{\gamma}$ and interleukin (IL)-2] and Th2 cytokine (IL-6 and IL-10) increased with increasing incubation time compared to the control in vitro. Enzymatic extract treatment promoted proliferation of splenocytes at concentrations of 100 and 200 mg/kg at 24 h in vivo. Secretion of $IFN-{\gamma}$ and IL-2 significantly increased in a dose-dependent manner at 24 h as well as induced higher production of IgG2a in serum. Natural killer cell activity was measured and tended to increase. In addition, complete blood cell counts increased in a dose-dependent manner. These results indicate that alginate oligosaccharides produced by crude enzyme from S. oneidensis PKA 1008 may have significant immune activities.

큰잎모자반에 함유되어 있는 다당류를 저분자화하기 위해 알긴산 분해 효소를 생산하는 Shewanella oneidensis PKA 1008의 조효소액을 첨가하여 저분자화하고 이의 면역증진능을 확인하였다. 먼저 저분자화됨을 확인하기 위하여 TLC를 실시한 결과 분해 24 h 이후부터 분해되기 시작하여 최종 60 h에서 dimers로 분해되었다. 그 후 비장세포에 큰잎모자반의 효소적 추출 분해물을 처리하여 in vitro에서 면역증진능을 확인한 결과 $IFN-{\gamma}$, IL-2, IL-6, IL-10의 경우 분해가 진행될수록 분비량이 많아졌다. 또한, 큰잎모자반의 효소적 추출 분해물을 2주 동안 마우스에 경구투여 한 결과, MTT assay 및 비장세포에서 분비되는 $IFN-{\gamma}$ 및 IL-2의 분비량의 경우 24 h 처리구에서 대조군보다 높은 수치로 농도 의존적으로 증가함을 나타내었다. IgG2a 분비량의 경우 24 h와 48 h 처리구에서 다소 증가하였으며, NK 세포의 경우 24 h 처리구에서 농도 의존적으로 활성이 증가하여 면역력을 증진시키는 결과를 보여주고 있다. 전혈을 이용하여 일반혈액검사의 경우 24 h 및 48 h 처리군에서 대조군보다 높은 수치를 나타내었다. 이상의 결과를 종합해볼 때 S. oneidensis PKA 1008 유래 조효소에 의한 큰잎모자반 효소적 추출 분해물이 면역 관련 세포증식률을 증가시키고 이에 따른 사이토카인의 분비량을 증가시키며 혈액 내 다양한 면역세포들의 수치를 증가시킴으로써 면역증진능에 탁월한 효과를 나타냄을 확인하였으나, 면역증진 효과에 기여하는 활성물질에 대해서는 밝혀지지 않아 그에 대한 추가적인 연구가 요구된다.

Keywords

References

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