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Fractionation of Exopeptidase from Viscera of Argentina Shortfin Squid, Illex argentinus

원양산 오징어(Illex argentinus) 내장으로부터 Exopeptidase의 분획

  • Kim, Hye-Suk (Division of Marine Life Science/ Institute of Marine Industry, Gyeongsang National University) ;
  • Kim, Jin-Soo (Division of Marine Life Science/ Institute of Marine Industry, Gyeongsang National University) ;
  • Heu, Min-Soo (Division of Marine Life Science/ Institute of Marine Industry, Gyeongsang National University)
  • 김혜숙 (경상대학교 해양생명과학부/해양산업연구소) ;
  • 김진수 (경상대학교 해양생명과학부/해양산업연구소) ;
  • 허민수 (경상대학교 해양생명과학부/해양산업연구소)
  • Published : 2008.08.30

Abstract

For the effective use of exopeptidase from squid viscera as food processing aids, the viscera of Argentina shortfin squid (Illex argentinus) were fractionated by various methods such as acetone treatment, ammonium sulfate treatment, anion exchange chromatography, and gel filtration. The positive exopeptidase fractions were obtained from the fraction II treated by cold acetone ($30{\sim}40%$, w/w), the fraction V by ammonium sulfate ($60{\sim}70%$ saturation), the fraction II (0.2 M NaCl) by anion exchange chromatography, and the fraction I ($30{\sim}50\;kDa$) by gel filtration. The specific activities of positive fractions from viscera of I llex argentinus against substrates were higher to LeuPNA than to ArgPNA. Total activity and recovery against LeuPNA of positive fraction by gel filtration were 1,867 U and 30.69%, respectively, which were the highest among those of positive fraction. The results suggested that the gel filtration chromatography method was the most efficient method for the fractionation of exopeptidase from viscera of Illex argentinus.

원양산 오징어 내장 조효소로부터 아세톤법, 황산암모늄법, 음이온교환 크로마토그래피법 및 겔 여과법을 이용하여 exopeptidase를 분획한 다음, 각 분획방법별 fraction들의 효소 활성(총활성 및 비활성), 정제도 및 회수율 등을 검토하여 최적 분획조건을 구명하고자 하였다. 아세톤 분획법의 경우 아세톤의 최종농도가 $30{\sim}40%$가 되도록 분획하는 것이, 황산암모늄 분획법의 경우 황산암모늄을 $60{\sim}70%$의 포화농도가 되도록 분획하는 것이, 음이온교환 크로마토그래피의 경우 0.2 M NaCl을 이용하여 분획하는 것이, 그리고, 겔 여과의 경우 분자량 $30{\sim}50\;kDa$ 범위의 것을 분획하는 것이 효율적이었다. 분획방법별 최적 활성 fraction간의 상호비교에서 LeuPNA 및 ArgPNA에 대한 총활성 및 회수율은 겔 여과가, 비활성 및 정제도는 황산암모늄에 의한 분획법이 가장 우수한 것으로 나타났다. 원양산 오징어 내장으로부터 탈 이온수로 조효소를 추출한 다음, 겔 여과로 분획한 exopeptidase의 식품가공소재로서 산업적 이용을 위해서는 겔 여과법의 특성을 기초로 하여 추출 조효소로부터 연속 및 대량처리를 위한 공정개발이 필요할 것으로 사료된다.

Keywords

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