해양에서 분리한 Bacillus sp. RH-5에 의한 사람 Low Density Lipoprotein(LDL) 산화에 대한 항산화제의 개발

Discovery of Antioxidant on Human Low Density Lipoprotein (LDL) by Bacillus sp. RH-5 Isolated from Marine Origin

Bacillus sp. RH-5로 배양한 배양액을 silica gel column (8.0cmX100cm)chromatography를 행하여 chloroform : methanol = 90 : 10인 항산화 분획층을 얻어 TLC 법에 의해 활성이 있는 물질을 분리하여 thiocyanate method로 활성을 측정하여 본 결과 Vit. E($10^{-2}$ mol)보다 좋았으며 BHA, BHT와는 비슷한 결과를 나타내었다. 한편 $5_\muM$ $CuSO_4$ 촉매 사람 Low Density Lipoprotein(LDL)의 산화에 항산화 활성이 알려진 Fraction 3의 band 4를 각각 100 및 200$\mu$g/$m\ell$ 농도로 $37^{\circ}C$에서 18시간 배양한 결과 LDL의 산화에 대한 억제효과가 좋았다. Fraction 3의 band 4를 HPLC로 정제한 후 IR. NMR 및 GC/MASS.로 확인한 결과 5-hydroxyndole로 확인되었다.

The aims of this studies were carried out to investigate the antioxidant activity on low density lipoprotein(LDL) using substances extracted from Bacillus sp. RH-5. The antioxidative substances produced extracellular in the culture broth by Bacillius sp. RH-S was obtained by elution of chloroform : methanol from silicagel column (80cm x100cm) chromatography. Band 4 eluted from fraction 3 by TLC was appeared at highest level of antioxidative activity using thiocyanate methed. Band 4 at a concentration of 100 or 200$\mu$g/$m\ell$ inhibited oxidation of LDL induced by the mouse transformed macrophage. According to IR. NMR or GC/MASS, the antioxidant substance was identified as 5-hydroxyindole.