摘要
为了解决目前藻红蛋白纯化方法过于复杂,处理量小等问题,采用“盐析→超滤→HA柱层析或DEAE-52柱层析”的工艺路线,建立了一种简化而高效的条斑紫菜藻红蛋白纯化方法。结果显示:1)对藻红蛋白纯度(以OD561/OD280表示)为0.365的条斑紫菜粗提物采用盐析法沉淀藻红蛋白时,得到藻红蛋白的纯度为0.45,回收率为90.53%;2)采用截留分子量为5万Da的超滤膜对藻红蛋白盐析物进行脱盐和出去小分子杂蛋白处理,得到的藻红蛋白的纯度为0.74,回收率为93.11%;3)进一步分别采用HA柱层析和DEAE-52柱层析纯化超滤液,藻红蛋白最高纯度分别为2.76和1.74,回收率分别为59.3%和56.81%。研究结果表明,采用盐析和超滤相结合的方法,可以较为方便而高效地得到食用级藻红蛋白,进一步通过HA柱层析,即可达到药用级的要求。
For overcoming some problems of present purification technology of phycoerythrin, such as too complicated process and too low yield, the method of ＂salting-out → ultrafiltration →hydroxyapatite column chromatography or DEAE-52 column chromatography＂ was used, an efficient and simple method on the purification of phycoerythrin from Porphyra yazoensis was established. Results show that ： 1） when the method of ammonium sulfate salting-out is applied to purify phycoerythrin from porphyra yazoensis crude extracts with an absorbance ratio （OD561/OD280） of 0. 365, a product with OD561/OD280 of 0. 45 and recovery yield of 67. 33% is obtained. 2） ultrafiltration with WM 50 000 Da is used for desalting and going out small molecule hetero-protein from salting-outyield of phycoerythrin, making purity （OD561/OD280） of phycoerythrin reach 0.74 and recovery rate reach 93.11%. 3） hydroxyapatite column chromatography and DEAE-52 column chromatography are further used, so the maximum purities of phycoerythrin reach 2.76 and 1.74 respectively, and the recovery rates reach 59.3%0 and 56.81% respectively. Research results indicate that salting-out combined with ultrafiltration is a high efficient method for obtaining edible-purity phycoerythrin. And if hydroxyapatite column chromatography is used further, medicinal-purity phycoerythrin can be obtained.
出处
《农业工程学报》
EI
CAS
CSCD
北大核心
2007年第2期249-253,共5页
Transactions of the Chinese Society of Agricultural Engineering
基金
江苏省高技术项目(BG2003319)
关键词
条斑紫菜
藻红蛋白
纯化
盐析
超滤
柱层析
Porphyra yazoensis
phycoerythrin
purification
salting-out
ultrafiltration
column chromatography
