ATPS法分离纯化Aspergillus sp.F044脂肪酶

研究了Aspergillussp.F044脂肪酶在双水相(ATPS)中的分配平衡,考察了PEG相对分子质量、PEG/磷酸盐质量分数、pH值和NaCl质量分数等5个重要因素对分配平衡的影响.发现PEG相对分子质量越小疏水作用越强,越有利于F044脂肪酶向上相分配,并且两相间电势差、氢键作用等对分配平衡有较大影响.试验得到的最优纯化条件为:PEG相对分子质量为600,质量分数为17.5%,磷酸盐质量分数为12.5%,w(NaCl)=0.5%,pH值为7.0,在此条件下脂肪酶分配系数达到7.1,纯化系数达到4.7,回收率达到91%.直线放大试验结果表明:该系统能够用于大量分离纯化Aspergillussp.F044脂肪酶,具有工业应用价值.

Separation and purification of Aspergillus sp.F044lipase by aqueous two phase system

The partition behavior of Aspergillus sp.F044 lipase in aqueous two phase system(ATPS) was examined.The results showed that five factors,including the molecular weight of polyethylene glycol,concentration of polyethylene glycol,concentration of phosphate salt,pH and concentration of NaCl have great influence on the partition behavior of the lipase in ATPS,and the less of molecule weight of polyethylene glycol,the more favorable for the lipase congregation into up phase because of the increased hydrophobicity.The optimized purification condition of Aspergillus sp.F044 lipase in ATPS is w(PEG600)=(17.5)?%,(w(phasphate salt)=)12.5?%,w(NaCl)=0.5?% and pH 7.0.Under these conditions,the partition coefficiency of the lipase is 7.1,purification fold is 4.7,and lipase yield is 91?%.The linear large-scaled trial indicated that the system could be applied to separation of commercial production of lipase and might have great prospect in light industry.