항산화물질을 생산하는 해양방선균 Streptomyces Sp. ACT-18의 동정 및 Electron Spin Resonance Spectrometry를 이용한 항산화활성

For the research of the natural marine antioxidant, an antioxidant-producing marine actinomycetes was isolated from sea water in Jeju coastal area. The strain was identified based on 16S rDNA sequencing, the morphology by a method of scanning electron microscopy, physiological and biochemical characteristics and cellular fatty acid analysis. The isolated strain ACT-18 was gram positive, aerobic, non-motile spores. Substrate mycelia are dark green and yellow gray aerial mycelia. The cell size of the strain was $0.5{\sim}1.0\;{\mu}m$. 16S rDNA sequence analysis showed that were Gram-positive bacteria grouped on Streptomyces sp. Results of cellular fatty acid analysis showed that major cellular fatty acids were $C_{15:0}$ anteiso (39.33%), $C_{16:1}$ cis 9 (11.96%), $C_{16:0}$ (13.08%) and $C_{17:0}$ anteiso (10.99%). The antioxidant activity of methanol extract from Streptomyce sp. ACT-18 was evaluated by measuring 1,1-diphenyl- 2-picrylhydrazyl (DPPH), hydroxyl, and alkyl radical scavenging activity using an electron spin resonance (ESR) spectrometer. DPPH radical scavenging activity of SBME (Streptomyces Broth Methanol Extract) A-18 was 46% at 0.1 mg/mL. Hydroxyl radical scavenging activity of SBME A-18 was 63% at 0.1 mg/mL. Alkyl radical scavenging activity of SBME A-18 was 39% at 0.1 mg/mL. 본 연구는 제주 연안 해수로부터 분리된 항산화물질을 생산하는 해양 방선균의 천연해양항산화물질에 관한 연구이다. 균주 ACT-18은 16S rDNA 염기서열, 전자현미경을 이용한 형태학적 분석, 생리학적, 생화학적 특성 및 세포 지방산분석을 통하여 동정되었다. 분리균주 ACT-18은 Gram 양성, 호기성, 비운동성 포자, 배지상에서 substrate mycelia의 색이 어두운 초록색, aerial mycelia는 노란색의 회색을 나타내였다. 세포사이즈는 약 $0.5{\sim}1.0\;{\mu}m$였다. 16S rDNA 염기서열 분석결과 Gram 양성박테리아인 Streptomyces 종들과 그룹을 형성하였다. 지방산분석결과 주요 지방산은 $C_{15:0}$ anteiso(39.33%), $C_{16:1}$ cis 9(11.96%), $C_{16:0}$ (13.08%)과 $C_{17:0}$ anteiso(10.99%)로 분석되었다. Streptomyce sp. ACT-18의 배양액 메탄올 추출물의 항산화 활성은 electron spin resonance (ESR) spectrometer에 의한 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, alkyl 라디컬 소거활성을 측정하여 나타내었다. 그 결과 배양액 메탄올 추출물의 DPPH 라디컬 소거활성은 0.1 mg/mL의 농도에서 46%의 라디컬 소거활성을 보였으며, Hydroxyl 라디컬 소거활성은 0.1 mg/mL에서 63%, alkyl 라디컬 소거활성은 0.1 mg/mL에서 39%의 소거 활성을 보였다.