title: Kinetics of inclusion body formation studied in intact cells by FT-IR spectroscopy.
abstract: 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.
authors.affiliation: Dipartimento di Biotecnologie e Bioscienze, Università di Milano Bicocca, Piazza della Scienza 2, 20126 Milan, Italy.
authors.forename: Diletta
authors.initials: D
authors.lastname: Ami
authors.forename: Antonino
authors.initials: A
authors.lastname: Natalello
authors.forename: Pietro
authors.initials: P
authors.lastname: Gatti-Lafranconi
authors.forename: Marina
authors.initials: M
authors.lastname: Lotti
authors.forename: Silvia Maria
authors.initials: SM
authors.lastname: Doglia
mesh_terms: 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
chemical_list: D011994:Recombinant Proteins; D008049:Lipase