重组毕赤酵母表达华根霉脂肪酶发酵液的絮凝除菌条件研究

摘要

在重组毕赤酵母表达华根霉脂肪酶的研究中,目的蛋白的提取收率是关键.由于产物脂肪酶是分泌在发酵液中,常用的方法是加入絮凝剂使茵体沉淀,再通过离心获取上清液中的目的蛋白.本研究改良了传统的絮凝剂配方,确定了絮凝剂的新配方:发酵液中加入2%的氯化钙以及磷酸氢二钠按摩尔比1:1混合液,500 mg/L聚丙烯酰胺,同时调节发酵液pH为6.0.结果表明,优化后的絮凝条件能有效的除茵并降低酶活损失.%Extraction rate of target protein is very important to the Rhizopus chinensis lipase expression in recombinant Pichia pastoris. Because the expressed lipase is an extracellular enzyme, the target protein is usually collected from the supernatant by centrifugation after adding flocculant into the broth to precipitate cells. This study improved the traditional flocculant formula. And the new formula of the flocculant was the following: 2% calcium chloride, 2% disodium hydrogen phosphate, 500 mg / L polyacrylamide and at pH 6. 0. The results showed that the optimized flocculant conditions were effective in removing cells and reducing activity loss.