坛紫菜藻红蛋白的规模化制备

Scale-up preparation of phycoerythrin from Porphyra haitanensis

为了优化坛紫菜粗提工艺以减轻纯化的压力,研究了我国主要经济红藻之一坛紫菜藻红蛋白大规模制备方法。实验采用"溶胀+组织捣碎"法破碎坛紫菜叶状体细胞,对比了多次硫酸铵梯度盐析对破碎液中藻红蛋白的影响,并进一步用羟基磷灰石层析制备藻红蛋白,最后对所得蛋白做了光谱和电泳鉴定。结果表明,经过4次盐析,藻红蛋白吸收光谱纯度达到0.9(A564/A280),每次的最佳盐析浓度分别为15%、50%、10%和40%;7 kg阴干紫菜经过4次盐析和1次羟基磷灰石层析后可获得507.82 mg藻红蛋白(A564/A280>3.2),为进一步研究规模高效分离纯化藻红蛋白提供了参考。

We developed large-scale preparation of phycoerythrin from Porphyra haitanensis,a main economic red algae in China.Firstly,P.haitanensis thallus was broken by using ＂swelling and smash＂ method.Then times of grads ammonium sulfate precipitation applied to the crude extraction were compared.Desalted solution was further purified with one-step chromatography using hydroxyapatite and properties on spectrum and molecular weight were identified finally.The results indicated that after four times of ammonium sulfate precipitation（15%,50%,10% and 40%）,the absorption spectrum purity of P.haitanensis achieved 0.9（A564/A280）,and 507.82 mg phycoerythrin（A564/A2803.2） was obtained from 7 kg fresh algae after further hydroxyapatite chromatography.This research provides a potential way for preparation of phycoerythrin in large sclae.