摘要
R-藻红蛋白的提取是其分离纯化的重要前提.因此,选择适合的提取方法是较为关键的步骤.以坛紫菜藻为材料,分别采用反复冻融法、液氮研磨法、细胞溶胀法、低浓度CaC l2提取法、SDS溶液提取法、纤维素酶提取法和超声波破碎法提取R-藻红蛋白,以期确定最佳的提取方法.研究结果表明:在反复冻融法中,以磷酸盐缓冲液为提取液,冷冻2 h,反复冻融6次,提取效果最佳(纯度0.631 6),且低温有利于保持蛋白活性;在低浓度CaC l2提取法中,0.05 mol/L CaC l2提取效果较好(纯度0.547 0),但会增加后续分离过程的难度且提取时间较长;在超声波破碎法中,以400 W功率超声20 m in,提取效果较好(纯度0.511 2)且时间短;其他提取方法虽然提取效果一般,但也有各自的优势,可依据实验要求选择合适的提取方法.
The extraction of R-phycoerythrin was an important prerequisite for its separation and purification.So it was the key step to select an appropriate extraction method.The R-phycoerythrin was extracted from Porphyra haitanensis by the methods of repeated freezing and melting,grinding with liguid nitrogen,dissolving-bulging,CaCl2 solution of low concentration,SDS solution extraction,cellulase extraction and ultrasonication,to determine the best method.The results showed that the effect was the best（purify 0.631 6） by 2 hours for freezing time and six cycles in repeated freezing and melting,and it had low temperature to maintain the protein activity.The effect of 0.05 mol/L CaCl2 extraction was better（purify 0.547 0）,but it would increase the difficulty of separation later,and the extraction time was longer.In ultrasonication,with 400 W and 20 min,the effect was better（purify 0.511 2） and the time was shorter.Although the effects of other methods were not very well,they had their own advantages.Therefore,an appropriate extraction method according to the experimental requirements could be selected.
出处
《哈尔滨商业大学学报（自然科学版）》
CAS
2011年第2期136-140,共5页
Journal of Harbin University of Commerce：Natural Sciences Edition
基金
国家自然科学基金(50808052)
哈尔滨市科技创新人才研究专项资金(2010RFLXS008)
关键词
坛紫菜藻
R-藻红蛋白
提取方法
纯度
porphyra haitanensis
R-phycoerythrin
extraction method
purify
