人溶菌酶的双水相萃取法分离

Separation of Human Lysozyme from Fermentation Broth of Pichia pastoris by Aqueous Two-Phase Extraction

摘要: 采用双水相体系直接从重组巴氏毕赤酵母发酵上清液中分离人溶菌酶,研究了体系中聚乙二醇(PEG)平均分子量,PEG、硫酸钠和氯化钠浓度,pH对人溶菌酶和总蛋白的分配系数、相体积比、萃取率和纯化因子的影响。结果表明:在PEG 4000、N a2SO4和N aC l质量分数分别为0.08、0.13、0.06,pH为5.6时,在室温下的双水相萃取率达96.63%,纯化因子为6.5。

Abstract: A new simple procedure using aqueous two-phase system was established for the separation of human lysozyme(HLZ) from the fermentation broth of Pichia pastoris.Effects of the relative molecular mass,concentration of polyethyleneglycol(PEG),Na_2SO_4 and NaCl,and pH value on the partition coefficient of HLZ,the volume ratio of total protein and phase,the yield and the purification factor were studied.Human lysozyme is recovered with a high yield in the top phase when PEG 4000,Na_2SO_4 and NaCl were in the two-phase system with the mass fraction of 0.08,0.13,0.06 respectively at pH 5.6.The purity of HLZ increases by the fractor of 6.5 with a yield of 96.63%.