The soluble expression of the protein using fusion partners: Thioredoxin, maltose binding human renin binding a comparison of ubiquitin, protein and NusA
The soluble expression of the protein using fusion partners: Thioredoxin, maltose binding human renin binding a comparison of ubiquitin, protein and NusA
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DOI：
10.1007/bf02940262
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发表时间：
2003-03-01
影响因子：
3.2
通讯作者：
Kim, BG
中科院分区：
文献类型：
--
作者：
Lee, C;Lee, SG;Kim, BG
人肾素结合蛋白（human renin binding protein, hRnBp）在大肠杆菌中过表达，但主要以包涵体形式存在，具有n -乙酰氨基葡萄糖-2- epimase活性。为了提高其溶解度和活性，用泛素（Ub）、硫氧还蛋白（Trx）、麦芽糖结合蛋白（MBP）和NusA作为融合体。融合蛋白的比较溶解度由高到低依次为：NusA、MBP、Trx、Ub。只有MBP融合没有明显降低hRnBp的活性，但增强了稳定性。折纸（DE3），允许更多的氧化。环境对大肠杆菌细胞质的影响，有助于提高其功能活性。
human renin binding protein (hRnBp), showing N-acetylglucosamine-2-epimerase activity, was over-expressed in E. coli, but was mainly present as an inclusion body. To improve its solubility and activity, ubiquitin (Ub), thioredoxin (Trx), maltose binding protein (MBP) and NusA, were used as fusion partners. The comparative solubilities of the fusion proteins were, from most to least soluble: NusA, MBP, Trx, Ub. Only the MBP fusion did not significantly reduce the activity of hRnBp, but enhanced the stability. The Origami (DE3), permitting a more oxidative. environment for the cytoplasm in E. coli, helped to increase its functional activity.