Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Purification and characterization of the extracellular alginate lyase from Streptomyces sp. MET 0515

Streptomces sp. MET 0515의 균체외 Alginate lyase의 정제 및 특성

  • Kim, Hyun-Kyoung (School of Biological Sciences and Technology, Chonnam National University) ;
  • Lee, Jae-Chang (Department of Dental Hygiene, Naju College) ;
  • Kang, Nam-Hyun (School of Biological Sciences and Technology, Chonnam National University) ;
  • Kim, Song-Hee (School of Biological Sciences and Technology, Chonnam National University) ;
  • Kim, Jong-Guk (Department of Microbiology, College of Nature Science, Kyungpook National University) ;
  • Chung, Ki-Chul (School of Biological Sciences and Technology, Chonnam National University)
  • 김현경 (전남대학교 생명과학기술학부) ;
  • 이재창 (나주대학 치위생과) ;
  • 강남현 (전남대학교 생명과학기술학부) ;
  • 김송희 (전남대학교 생명과학기술학부) ;
  • 김종국 (경북대학교 미생물학과) ;
  • 정기철 (전남대학교 생명과학기술학부)
  • Published : 2007.05.25
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Abstract

We isolated a new extracellular alginate lyase-producing microorganism, which displayed alginate-depolymerizing activity in plate assays, from coastal soils in Wando, Jeollanam-do, Korea. This alginate-depolymerizing bacterium belonged to the genus Streptomyces and it was named Streptomyces sp. MET 0515. An extracellular alginate lyase(ALY1) secreted by Streptomyces sp. MET 0515, was purified to homogeneity by a combination of acetone precipitation, anion-exchange chromatography (Q-Sepharose and DEAE-Sepharose) and Sephacryl S-200 HR gel filtration chromatography. Its molecular mass was 26 kDa as determined by SDS-PACE analysis. The enzyme had an optimal temperature of $70^{\circ}C$ for its activity, and was most active at pH 7.5. The thermal and pH stability were $0-50^{\circ}C$, and pH 6.0-9.0, respectively. The enzyme activity was stimulated by 1mM $Mn^{2+}$, and inhibited by 1mM $Fe^{3+}$, 1mM EDTA and 1mM $Zn^{2+}$. Preliminary analysis of substrate specificity showed that this alginate lyase had activity on both poly-alpha 1,4-L-guluronate and poly-beta 1,4-D-mannuronate in the alginate molecule.

알긴산분해효소(EC 4. 2. 2. 3)를 균체외로 생산하는 새로운 방선균을 전라남도 완도의 연안토양으로부터 분리하고 이 균주를 Streptomyces sp. MET 0515라 명명하였다. 이 균주가 생산하는 균체외 알긴산분해효소를 아세톤 침전, 음이온 교환 크로마토그래피(Q-Sepharose and DEAE-Sepharose), 젤 크로마토그래피(Sephacryl S-200 HR gel filtration chromatography)를 이용하여 정제하였다. 이 효소의 최적 활성 온도과 pH는 각각 $70^{\circ}C$와 pH 7.5이고, 온도 안정성은 $0-50^{\circ}C$, pH 안정성은 pH 6.0-9.0였다. 이 효소는 $Mn^{2+}$ 이온 첨가시에는 활성이 증가하였으며 1mM $Fe^{3+}$, 1mM EDTA, 1mM $Zn^{2+}$ 이온 첨가시에는 활성이 억제되었다. 또한 이 효소는 poly-alpha 1,4-L-guluronate와 poly-beta 1,4-D-mannuronate두 종류의 기질에 대하여 활성을 나타냈다. 따라서 본 효소는 다른 미생물 기원의 효소와 비교해 볼 때 Streptomyces sp.가 생산하는 첫번째 효소로 인정되었다.

Keywords

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