High-pressure infrared spectroscopic study of human proinsulin gene expression in live escherichia coli cells

Abstract

Infrared spectra of E. coli strain JM103 and transformants which overproduced recombinan proinsulin have been measured as a function of pressure up to 38 kbar. It is the first time that high-pressure infrared spectra of live bacteria have been successfully measured. In ambient conditions, spectra of the host strain JM103 and the transformants are generally identical. However, under pressure, distinct shifting pattern can be observed in specific spectral parameters of transformants, presumably due to accumulation of proinsulin in form of cytoplasmic inclusion bodies. In particular, the pressure-induced frequency shift of the amide III band (1235 cm −1) in the proinsulin-producing transformants is much smaller than in the host JM103. This pressure effect can potentially be an efficient approach to monitor maximum gene expression in microorganisms. Contrary to predictions based on model system, the pressure-induced denaturation and the sharp transition from disordered liquid crystalline state to the ordered gel state commonly observed in the aqueous solution of protein and aqueous bilayer dispersion of lipids, respectively, do not occur in the bacterial proteins and cell membrane of E. coli.