食品科学

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两步盐析联合双水相萃取提取纯化蓝藻中藻蓝蛋白

张发宇,赵冰冰,陈 裕,袁梦媛,汪家权   

  1. 合肥工业大学资源与环境工程学院,安徽 合肥 230009
  • 出版日期:2015-11-25 发布日期:2015-12-03
  • 通讯作者: 汪家权
  • 基金资助:

    “十二五”国家科技重大专项(2012ZX07103-004)

Extraction and Purification of Phycocyanin by the Combined Use of Two-Step Salt Precipitation and Aqueous Two-Phase Extraction from Blue Algae

ZHANG Fayu, ZHAO Bingbing, CHEN Yu, YUAN Mengyuan, WANG Jiaquan   

  1. School of Resources and Environmental Engineering, Hefei University of Technology, Hefei 230009, China
  • Online:2015-11-25 Published:2015-12-03
  • Contact: WANG Jiaquan

摘要:

以巢湖新鲜蓝藻为处理对象,采取冻融破壁的方式获取藻蓝蛋白的粗提液,采取两步盐析联合双水相萃取的方法提取纯化藻蓝蛋白。运用0.618法选点实验研究了两步盐析中(NH4)2SO4饱和度对提取纯化影响,构建了聚乙二醇(polyethylene glycol,PEG)-(NH4)2SO4的双水相体系,综合考虑PEG相对分子质量、PEG质量分数、(NH4)2SO4质量分数、pH值和藻蓝蛋白质量浓度对提取纯化藻蓝蛋白的影响。结果表明:室温25 ℃条件下,一步盐析时(NH4)2SO4的最佳饱和度为21%,二步盐析时(NH4)2SO4的最佳饱和度为30%。两步盐析后得到的藻蓝蛋白构建成PEG-(NH4)2SO4的双水相体系后,当PEG相对分子质量1 000、PEG 1 000质量分数10%、(NH4)2SO4质量分数25%时,藻蓝蛋白的纯度可达3.45。

关键词: 两步盐析, 双水相萃取, 提取纯化, 藻蓝蛋白

Abstract:

The crude extracts of phycocyanin from fresh blue algal were obtained by freeze-thaw method. The purification of
phycocyanin was performed by the combined use of two-step salting-out precipitation and aqueous two-phase extraction. The
effect of different saturation degrees of (NH4)2SO4 in two-step salt precipitation on purification efficiency was investigated
using the 0.618 method, and the aqueous two-phase system of polyethylene glycol (PEG)-(NH4)2SO4 was established after
comprehensively considering the effect of PEG molecular weight, PEG mass fraction, (NH4)2SO4 mass fraction, pH and
phycocyanin mass concentration on purification efficiency. The results showed that the optimum (NH4)2SO4 saturation was
21% in the first step and 30% in the second step salt precipitation at 25 ℃. Using the aqueous two-phase system containing
10% PEG 1 000 and 25% (NH4)2SO4, the purity of purified phycocyanin was 3.45.

Key words: two-step salting-out precipitation, aqueous two-phase extraction, extraction and purification, phycocyanin

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