생육시기 및 부위별 고추의 항산화력 및 항암 Lunasin peptide의 동정

Abstract

부위간의 DPPH free radical scavenging 활성은 과육은 종자 보다 높았고 생육시기간에는 후기가 초기에 비하여 높았고, Xanthine oxidase 억제활성은 종자와 과육에 비하여 매우 높았고 동정된 항 산화물은 대부분이 지방산 및 페놀성 화합물로, 과육 및 종자가 성숙 될수록 1-eicosanol, palmitic acid, linoleic acid, linolenic acid 등의 지방산 및 benzonitrile의 함량이 증가되었다. 시료 채취 시기간의 POD활성은 과육은 성숙중기에 높았으나, 종자는 성숙이 경과될수록 낮았다. SOD 활성은 과육은 생육이 경과할수록 증가하였으나, 종자는 성숙할수록 낮아졌다. 고추종자 단백질은 western blot에 의하여 lunasin이 동정되었고, 개화 후 5주부터 lunasin 생합성 되었다. 고추 종자내의 100nM lunasin 펩타이드는 종양유발 유전자가 전이된 <TEX>$2\~12$</TEX> 세포의 콜로니 형성을 억제시켰으며, 결과적으로 고추 종자내에는 암 예방적 특성을 갖는 lunasin peptide가 존재한다. The non-enzymatic anti-oxidants and lunasin peptide from the extracts of the pepper were examined in order to utilize the discovery in natural products as cancer chemopreventive agents. The DPPH (1,1-diphenyl-2-picryl-hydrazyl) free radical scavenging activity on the fruit parts of the pepper was higher than that of the seed, but the difference was low. The Inhibition activity of xanthine/ xanthine oxidase in extracts of the seed was higher than that of the fruit and that of the seed on 20 days after flowering was the highest at the growing period. These were identified as fatty acids and phenolic compounds such as 1-eicosanol, palmitic acid, linoleic acid, linolenic acid and benzonitrile. The contents of fatty acids and phenolic compounds increased according to the time passing at the growing period. Peroxidase (POD) activity of the fruit at middle stage was high than that of other growing stages and that of the seed was the highest at later growing period. Though superoxide dismutase (SOD) activities in fruit were hish by passage of Slowing stage, the activity in seed was low. Lunasin was searched from seeds of the peppers by coomassie blue staining and western blot among them and we just found lunasin peptide from extracted protein of the pepper by western blot. In addition, we observed the contents of lunasin after flowering and confirmed to appear the lunasin at 35 days after flowering. We confirmed that lunasin is complex protein of maturing seeds. 100nM lunasin peptide in pepper showed inhibition effect on colony formation in <TEX>$2\~12$</TEX> cells.