Purification and Characterization of a Carotenoprotein from Penaeus orientalis

대하(P. orientalis)로부터 분리정제한 Carotenoprotein의 특성에 관한 연구

  • Published : 19940800

Abstract

The isolation, purification and characterization of a carotenoprotein from the carapace of Pnaeus orientalis were investigated. The carotenoprotein was purple with broad λmax between 480, 409, 318 and 280 nm. Apparent structures were estimated by using X-ray diffractometry and scanning electron microscope, respectively. The molecular weight of the carotenoprotein complex had been determined by GPC and PAGE. The heavier complex, designated the $\alpha$-form (M.W = 170 KDa), was dissociated to a major subunit, $\beta$-form (M.W = 42 KDa). SDS-PAGE of $\alpha$-form showed apparently oligomeric pattern, and also $\beta$-form gave two polypeptides corresponding to 22 KDa and 19 KDa, respectively. The amino acid of the two proteins $({\alpha}-and\;{\beta})$-form, lipid and free fatty acid compositions were described. The prosthetic groups of the carotenoprotein were confirmed by TLC, IR, $^1H$-NMR, MS and various organic reactions as astaxanthin, astaxanthin monoester and astaxnathin diester.

양식산 새우(P.orientalis)로부터 0.05M 인산염 완충용액(pH 7.0)으로 carotenoprotein을 추출하고 DEAE-셀룰로오스, 반포화 황산암모늄 친전 및 GPC로 $\alpha$-form과 ${\beta}$-form 두 종류의 carotenoprotein을 분리 정제하였다. 최대 흡수파장은 ${\lambda}_{max}$= 480, 409, 318 및 280 nm였다. SEM과 XRD로 분자구조를 관찰하였으며, GPC와 PAGE로 측정한 분자량은 $\alpha$-form이 170KDa으로 분리되는 불균일한 이합체 구조였다. carotenoprotein 복합체의 아미노산, 중성지질, 인지질 및 유리지방산 조성을 분석하였고, 여러 가지 유기화학 반응과 UV/Vis,IR, $^1H$-NMR, MS 등으로 carotenoids분자구조를 측정한 결과 lipid-protein의 보호그룹으로서는 astaxanthin과 그 ester임이 확인되었다.

Keywords

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