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首页> 外文期刊>Bioprocess and Biosystems Engineering >Investigation of chromatography and polymer/salt aqueous two-phase processes for downstream processing development of recombinant phenylalanine dehydrogenase
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Investigation of chromatography and polymer/salt aqueous two-phase processes for downstream processing development of recombinant phenylalanine dehydrogenase

机译：色谱法和聚合物/盐水溶液两相法研究重组苯丙氨酸脱氢酶的下游工艺开发

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摘要

This work presents a comprehensive study between the polymer/salt aqueous two-phase systems (ATPS) and chromatography process for downstream processing of recombinant Bacillus badius phenylalanine dehydrogenase (PheDH). First, the partitioning behavior of recombinant PheDH in polyethylene glycol (PEG)/ K_2HPO_4 ATPS was examined. For comparative purpose, a classical chromatographic protocol was performed as well. Investigation of chromatography and ATPS procedures revealed that the ATPS comprising of 9% (w/w) PEG-6000, 16% (w/w) K_2HPO_4 and 16% (w/w) KCl with pH of 8.0, volume ratio (V_R) of 0.25, temperature of 25 ℃ and 40% (w/w) cell lysate ensured the most favorable approach for PheDH downstream process. A specific activity of 4,231.4 U/mg, a yield of 96.7% and a recovery of 162.0% were obtained. Furthermore, the shorter process time (4 vs. 48 h) and the lower total cost (4 vs. 20 €) were additionally features that confirmed the suitability of proposed technique.
机译：这项工作提出了聚合物/盐水溶液两相系统（ATPS）与色谱过程之间的综合研究，以用于重组巴斯德毕赤酵母苯丙氨酸脱氢酶（PheDH）的下游加工。首先，研究了重组PheDH在聚乙二醇（PEG）/ K_2HPO_4 ATPS中的分配行为。为了比较，还进行了经典的色谱操作。色谱和ATPS程序的研究表明，ATPS包含9％（w / w）PEG-6000、16％（w / w）K_2HPO_4和16％（w / w）KCl，pH值为8.0，体积比（V_R） 0.25℃，25℃的温度和40％（w / w）的细胞裂解液确保了PheDH下游工艺的最佳方法。获得的比活性为4,231.4 U / mg，产率为96.7％，回收率为162.0％。此外，更短的处理时间（4 vs. 48 h）和更低的总成本（4 vs. 20€）是另外的特征，证实了所提出技术的适用性。

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