Purification and Characterization of Phospholipase D from Actionmycetes KF923

방선균 KF923이 생산하는 Phospholipase D의 정제 및 특성

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

Abstract

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.

Keywords

Purification;characteristics;phospholipase D;Actinomycetes

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