Panek Anna, Pietrow Olga, Filipkowski Paweł, Synowiecki Józef
Department of Food Chemistry, Technology and Biotechnology, Faculty of Chemistry, Gdansk University of Technology, Gdańsk, Poland.
Acta Biochim Pol. 2013;60(2):163-6. Epub 2013 Jun 6.
Two recombinant trehalose synthases from Deinococcus geothermalis (DSMZ 11300) were compared. A significant influence of the artificial polyhistidine tag was observed in protein constitution. The recombinant trehalose synthase from D. geothermalis with His₆-tag has a higher Km value of 254 mM, in comparison with the wild-type trehalose synthase (Km 170 mM), and displayed a lower activity of maltose conversion when compared to the wild type. Moreover, differences in properties like temperature, pH, thermal- and pH-stability were observed. Presence of the histidine tag caused a decrease of thermal resistance in case of trehalose synthase with His₆-tag. These data confirmed a suggestion that the introduction of the histidine domain produces in some seldom cases undesirable changes in the protein.
对来自嗜热栖热放线菌（DSMZ 11300）的两种重组海藻糖合酶进行了比较。观察到人工多聚组氨酸标签对蛋白质组成有显著影响。与野生型海藻糖合酶（Km为170 mM）相比，带有His₆标签的嗜热栖热放线菌重组海藻糖合酶的Km值更高，为254 mM，并且与野生型相比，其麦芽糖转化活性较低。此外，还观察到温度、pH值、热稳定性和pH稳定性等性质的差异。组氨酸标签的存在导致带有His₆标签的海藻糖合酶的耐热性降低。这些数据证实了一种观点，即组氨酸结构域的引入在某些罕见情况下会导致蛋白质产生不良变化。
