工业微生物2017，Vol.47Issue(4)：43-48,6.DOI:10.3969/j.issn.1001-6678.2017.04.007
3′,5′-二磷酸核苷酸酶在亲和双水相胶束系统中分配行为研究
Studies on partitioning behavior of 3′,5′-bisphosphate nucleotidase in affinity-based aqueous two-phase micellar system
摘要
Abstract
In order to improve the specificity of ATPMS,a novel affinity-based aqueous two-phase micellar system (ATPMS) was constructed by introducing a metal-chelated surfactant Triton X-114-IDA-Cu(II) (TX-Cu(II)) into an aqueous solution of hydrophobically modified ethylene oxide polymer (HM-EO),and the possibility of this HM-EO/TX-Cu(II) system for separating histidine-tagged proteins was investigated by the affinity partitioning of recombinant protein 3′,5′-bisphosphate nucleotidase (YND).It was found that in the absence of TX-Cu(II),the YND tended to partition into the micelle-poor phase,where it experienced fewer excluded-volume effects.However,with the addition of TX-Cu(II),the YND was partitioned preferentially to the micelle-rich phase due to affinity binding and was stable in the system.The pH of the solution could affect the partitioning behavior of YND,and the optimum condition for the extraction of YND was pH 9.0.With increasing NaCl concentration,more protein impurities were removed to the micelle-poor phase,but the partition coefficient of YND fluctuated slightly.Finally,the YND was attained in the micelle-rich phase with a recovery yield of 65.8% under the conditions of HM-EO 2.5%,TX-Cu(II) 0.125%,pH 9.0 and 50 mmol/L NaCl.The results indicated that the new affinity-based HM-EO/TX-Cu(II) system had high partitioning performance which was promising for effectively separation of the histidine-tagged proteins.关键词
双水相胶束系统/金属螯合表面活性剂/亲和分配/3′,5′-二磷酸核苷酸酶Key words
aqueous two-phase micellar system/metal-chelated surfactant/affinity partitioning/3′,5′-bisphosphate nucleotidase引用本文复制引用
刘耀,周鑫鑫,邓刚..3′,5′-二磷酸核苷酸酶在亲和双水相胶束系统中分配行为研究[J].工业微生物,2017,47(4):43-48,6.基金项目
国家自然科学基金(No.21006098). （No.21006098）
