葛仙米藻胆蛋白的提取工艺优化及其纯化研究

Research on Optimization of Phycobiliprotein Extraction from Nostoc Sphaeroids Kutzing and the Purification of Phycobiliprotein

以葛仙米为原料,研究葛仙米藻胆蛋白的超声辅助提取工艺。以藻胆蛋白提取率为评价指标,对提取工艺中葛仙米粉粒径数、超声功率、处理方法、液料比、提取时间、提取温度共6个因素进行单因素试验,并结合响应面分析对提取工艺进行优化。结果表明:采用超声辅助处理方法(超声功率700 W),当葛仙米粒径数120目~200目、液料比58∶1(m L/g)、提取温度42℃、提取时间6 h时,葛仙米藻胆蛋白的提取率达到最大为26.13%,达到预测值的99.54%。采用一步离子交换树脂层析法纯化藻蓝蛋白(Phycocyanin,PC)和藻红蛋白(Phycoerythrin,PE),经聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)、凝胶过滤色谱(Gel filtration chromatography,GFC)、紫外可见吸收光谱(Ultraviolet-visible absorption spectroscopy,UV-Vis),藻蓝蛋白纯度达到93%,藻红蛋白纯度83.5%。

bThe ultrasonic -assisted extraction of phycobiliprotein from the raw material,nostoc sphaeroidskutzing,was investigated in this work. The single factor test investigated parameters of the extraction processincluded dry nostoc sphaeroids kutz powder size,ultrasonic irradiation power,treatment method,liquid-to-solid ratio,extraction time and extraction temperature,to evaluate the extraction rate of alga protein. Theoptimized extraction conditions were determined with the aid of the response surface analysis. Results indicatedthat the extraction yield of nostoc sphaeroids kutz phycobiliprotein was up to 26.13%,reaching the predictedvalue of 99.54%,under the optimized conditions,nostoc sphaeroids kutz powder size at 120 to 200 mesh,ultrasonic irradiation power 700W,ultrasonic -assisted treatment,liquid -to -solid ratio at 58 ： 1（mL/g）,extraction temperature 42 ℃, and extraction time 6 h. Phycocyanin and phycoerythrin were purified by one-stepion exchange resin chromatography. By sodium dodecyl sulfate polyacrylamide gel electrophoresis, Gel filtrationchromatography and Ultraviolet-visible absorption spectroscopy,the purities of the purified PC and PE were93 % and 83.5%, respectively.