방선균 KF923이 생산하는 Phospholipase D의 정제 및 특성

Purification and Characterization of Phospholipase D from Actionmycetes KF923

  • 곽보연 (한국식품개발연구원 식품기능연구본부) ;
  • 윤석후 (한국식품개발연구원 식품기능연구본부) ;
  • 김창진 (한국생명공학연구원) ;
  • 손동화 (한국식품개발연구원 식품기능연구본부)
  • 발행 : 2004.03.01

초록

국내 6개 지역에 걸쳐 채취한 토양 63점으로부터 1,000여 종의 방선균주를 분리하였다. 이들 균주의 배양 상징액 중 PLD의 생산이 양호한 균주를 탐색한 결과, 분해활성이 0.3U/$m\ell$이상인 131 균주를 일차로 선발하였다. 이중 전이 활성이 20% 이상인 균주는 23개였으며, 최종적으로 분해 및 전이활성이 가장 우수한 균주(방선균 KF923)를 선발하였다. 방선균 KF923 균주는 P 배지하에서 발효조 배양시 배양 48시간 후에 최고의 분해활성을 나타내었고(13U/$m\ell$) 전이활성은 95%로 나타났다. 방선균 KF923 균주가 생산한 PLD를 정제하여 비활성이 567U/mg의 PLD를 얻었다. 정제된 PLD는 분자량이 약 55kD으로 나타났고, 효소반응의 최적 pH는 6.0, 최적온도는 $60^{\circ}C$였으며, 효소의 안정성에 미지는 pH 및 온도의 영향은 pH 8.0부근 및 $40^{\circ}C$ 이하에서 가장 안정하였다. 또한, 특별한 금속이온의 요구성은 없는 것으로 나타났다. 방선균 KF923 균주가 생산한 PLD는 산업화에 충분히 활용 가능하며, 이를 위하여 효소의 대량생산 및 효소반응을 위한 공정개발 등의 연구가 필요하다고 생각한다.

In order to screen microorganisms producing phopholipase D (PLD) had high transphosphatidylation activity, about 1,000 Actinomycetes strains were isolated from the 63 soil samples, collected over 6 local area in Korea. When the hydrolytic activity in the supernatant was determined, 131 strains produced PLD more than 0.3U/$m\ell$. Among 131 culture broths tested, 23 ones had transphosphatidylation activity higher than 20% and finally one strain (Actinomycetes KF923), which had highest hydrolytic and transphophadylation activity, was selected. Actinomycetes KF923 showed the highest hydrolytic activity (13U/$m\ell$) and phosphatidylation activity (95%) after 48 h fermentation using the P medium (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, $CaCO_3$ 0.4%, pH 7.2). PLD was purified from the culture broth of Actinomycetes KF923 and the specific activity of purified PLD was 567U/mg. The molecular weight of PLD was about 55kD and the optimum pH and temperature were pH 6.0 and $60^{\circ}C$, respectively. The stability of PLD toward pH and temperature were high around pH 8.0 and below $40^{\circ}C$ Special metal ions were not necessary to the PLD activity.

키워드

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