Kinetics of inclusion body formation studied in intact cells by FT-IR spectroscopy.
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.
Ami
Diletta
D
Dipartimento di Biotecnologie e Bioscienze, Università di Milano Bicocca, Piazza della Scienza 2, 20126 Milan, Italy.
Natalello
Antonino
A
Gatti-Lafranconi
Pietro
P
Lotti
Marina
M
Doglia
Silvia Maria
SM
D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D002479:Inclusion Bodies / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared
D011994:Recombinant Proteins; D008049:Lipase