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

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

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.

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.