Heat-induced translocation of cytoplasmic beta-galactosidase across inner membrane of Escherichia coli

Biotechnology Progress
Hiroshi UmakoshiY Matsumura

Abstract

The behaviors of heat-induced translocation of cytoplasmic beta-galactosidase to periplasm across the inner membrane of Escherichia coli cells were investigated in order to apply such phenomena to the process for production and separation of intracellular biomolecules. The heat stress was found to induce translocation of cytoplasmic beta-galactosidase (beta-gal) together with reduction of the amounts of intracellular soluble proteins and formation of their inactive aggregates. The translocation of beta-gal was then analyzed using (a) the location factor of beta-gal (LFG), which meant enzyme location in the cells and could be determined from the kinetic analysis of enzyme release process, and (b) the percentage of beta-gal activity in periplasm after solublizing the outer membrane of E. coli cells by lysozyme/EDTA treatment. The LFG values were maximized when cells were stressed at the temperature of 42-47 degrees C. From the results on the surface properties of both beta-gal and cell membrane under the heat stress, it is suggested that (1) the conformational change of cytoplasmic oligomeric beta-gal to the partially dissociated and/or unfolded state with higher local hydrophobicity, (2) the increase in membrane fluidity of inne...Continue Reading

Citations

Jul 8, 2005·Applied Microbiology and Biotechnology·F R Schmidt
Oct 20, 2005·Journal of Bioscience and Bioengineering·S MoritaR Kuboi
Oct 20, 2005·Journal of Bioscience and Bioengineering·Seiichi MoritaRyoichi Kuboi
Dec 18, 2001·Biotechnology and Bioengineering·B Balasundaram, A B Pandit

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