Study on extraction and purification of C-phycocyanin in Synechococcus sp.PCC 7002
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摘要: 本文研究了聚球藻7002藻蓝蛋白的分离纯化工艺,为提高聚球藻7002藻蓝蛋白的纯度和提取率提供一定技术指导。采用组织捣碎法、超声波法、反复冻融法对细胞进行破碎提取藻蓝蛋白,比较了不同破壁法提取藻蓝蛋白的纯度和总蛋白提取率,采用响应面法确定了最佳破壁条件,通过硫酸铵盐析、羟基磷灰石层析法提纯藻蓝蛋白,对结果进行SDS电泳鉴定。实验显示反复冻融法破壁效果最佳,响应面实验确定了最佳破壁条件为菌体浓度3.53 mg/m L、冻融次数6次、解冻温度37℃、最佳盐析浓度为硫酸铵55%,经羟基磷灰石层析后藻蓝蛋白纯度可达3.79。最终本实验得到了分离纯化聚球藻7002藻蓝蛋白的最佳工艺方法。Abstract: In order to improve the purity and production of C-phycocyanin( C-PC) in Synechococcus sp.PCC 7002,extraction and purification of C-phycocyanin in Synechococcus sp.PCC 7002 was investigated. Tissue-trituration method,cycle of freezing and thawing method and ultrasonic method were applied and compared to the effection on the purity and production of C-phycocyanin. Response surface methodology( RSM) was used to optimize the best cell- disruption conditions. Futher purification was performed by means of ammonium sulphate precipitation and hydroxyapatite( HA) chromatography. The purified protein was examined through SDS-PAGE. The results showed that cycle of freezing and thawing method was the optimal cell-disruption method by the RSM experiment,the best cell-disruption conditions were that the concentration of bacteria was 3.53 mg / m L,the cycles of freezing and thawing was 6 times,the thawing temperature was 37 ℃,the best concentration of salting out was55% ammonium sulfate,the purity of C-phycocyanin reached 3.79 through hydroxyapatite chromatography. The optimal method of extraction and purification of phycocyanin from Synechococcus sp.PCC 7002 was achieved.
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