建立了一种分离方法,以依次分离和回收三种源自糖的抗炎成分,藻胆蛋白,和红色藻类dulse(Palmariapalmata)的热空气干燥的thalli的叶绿素。开发的过程包括三个步骤,不使用有机溶剂。在步骤I中,通过用多糖降解酶破坏干燥的thalli的细胞壁来分离糖,通过沉淀其他成分获得富含糖的提取物(E1),同时通过酸沉淀洗脱。在步骤二,将步骤I的残留悬浮液用嗜热菌蛋白酶消化以获得藻胆蛋白衍生肽(PPs),并且通过使用酸沉淀分离其它提取物获得富含PP的提取物(E2)。在步骤三,通过加热残留物获得溶解的叶绿素,是酸沉淀的,中和,并重新溶解以浓缩富含叶绿素相关成分(Chls)的提取物(E3)。这三种提取物通过脂多糖(LPS)刺激的巨噬细胞抑制炎症细胞因子的分泌,确认序贯程序对任何提取物的活性没有负面影响。E1、E2和E3富含糖,PPs,和Chls,分别,表明抗炎成分通过分离方案被有效地分级和回收。
A separation process was established to sequentially fractionate and recover three anti-inflammatory components derived from sugars, phycobiliprotein, and chlorophyll from the hot-air-dried thalli of the red alga
dulse (Palmaria palmata). The developed process consisted of three steps, without the use of organic solvents. In Step I, the sugars were separated by disrupting the cell wall of the dried thalli with a polysaccharide-degrading enzyme, and a sugar-rich extract (E1) was obtained by precipitating the other components, which were simultaneously eluted by acid precipitation. In Step II, the residue suspension from Step I was digested with thermolysin to obtain phycobiliprotein-derived peptides (PPs), and a PP-rich extract (E2) was obtained by separating the other extracts using acid precipitation. In Step III, solubilized chlorophyll was obtained by heating the residue, which was acid-precipitated, neutralized, and re-dissolved to concentrate the chlorophyll-related components (Chls)-rich extract (E3). These three extracts suppressed inflammatory-cytokine secretion by lipopolysaccharide (LPS)-stimulated macrophages, confirming that the sequential procedure had no negative effects on the activities of any of the extracts. The E1, E2, and E3 were rich in sugars, PPs, and Chls, respectively, indicating that the anti-inflammatory components were effectively fractionated and recovered through the separation protocol.