Ami Diletta, Natalello Antonino, Gatti-Lafranconi Pietro, Lotti Marina, Doglia Silvia Maria
Dipartimento di Biotecnologie e Bioscienze, Università di Milano Bicocca, Piazza della Scienza 2, 20126 Milan, Italy.
FEBS Lett. 2005 Jun 20;579(16):3433-6. doi: 10.1016/j.febslet.2005.04.085.
The aggregation of a recombinant lipase as inclusion bodies (IBs) was studied directly within intact Escherichia coli cells by FT-IR microspectroscopy. Through this approach, it was possible to monitor in real time the different kinetics of IB formation at 37 and 27 degrees C, in excellent agreement with the results of the SDS-PAGE analysis. Furthermore, insights on the residual native-like structure of the expressed protein within IB--both isolated and inside cells--were obtained by the secondary structure analysis of the Amide I band in the IB FT-IR spectra.
通过傅里叶变换红外光谱（FT-IR）显微技术，直接在完整的大肠杆菌细胞内研究了重组脂肪酶作为包涵体（IBs）的聚集情况。通过这种方法，可以实时监测在37℃和27℃下IB形成的不同动力学，这与十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析结果高度一致。此外，通过对IB的傅里叶变换红外光谱中酰胺I带的二级结构分析，获得了关于分离的和细胞内IB中表达蛋白残留的类天然结构的见解。
