摘要
为了研究极端嗜热细菌腾冲嗜热厌氧杆菌葡聚糖磷酸化酶(Tte-GlgP)的性质,将用大肠杆菌BL21DE3表达的Tte-GlgP进行了纯化,分别用SDS-PAGE电泳法及毛细管电泳法测定了分子量及等电点,结果表明,Tte-GlgP的分子量约为64kDa,等电点约为6.2,与推测的结果一致,说明Tte-GlgP在大肠杆菌体内可能得到了正确的折叠;WESTERN杂交试验的结果表明,Tte-GlgP在腾冲嗜热厌氧杆菌体内有表达,且终浓度为1.5%的麦芽糖能轻度诱导Tte-GlgP在腾冲嗜热厌氧杆菌体内的表达,而终浓度为1.5%的葡萄糖则轻度抑制Tte-GlgP在腾冲嗜热厌氧杆菌体内的表达,说明Tte-GlgP在腾冲菌体内可能参与了碳水化合物的代谢。本研究为进一步认识和应用Tte-GlgP提供了依据。
To investigate the characteristics of a glucan phosphorylase from Thermoanaerobacter tengcongens （Tte-GlgP）, Tte-GlgP expressed in E. coli was purified, and its molecular weight and pI were determined by SDS-PAGE and capillary electrophoresis, respectively. It was found that the molecular weight and pI of the recombinant Tte-GlgP were about 64 kDa and 6.2, respectively. These results suggested that the Tte-GlgP might correctly folded in E. coli. The results of Western. hybridize showed that the glgP gene was expressed in T. tengcongensis. 1.5% maltose could slightly induce the expression level of Tte-GlgP, whereas 1.5% glucose slightly inhibited its expression level in T. tengcongensis. These results suggested that Tte-GlgP might be involved in the carbohydrate metabolism in T. tengcongensis. This article could provide some useful reference to deeply understand and further use Tte-GlgP.
出处
《中国酿造》
CAS
北大核心
2009年第4期74-76,共3页
China Brewing
