Role of methoxypolyethylene glycol on the hydration, activity, conformation and dynamic properties of a lipase in a dry film.

Biotechnology and Bioengineering
F SecundoGiorgio Mazzola

Abstract

A combined approach based on the use of ATR-FT/IR and steady-state fluorescence spectroscopy allowed to shed light on the effects of the additive methoxypolyethylene glycol (MePEG) on the hydration, conformation and dynamic properties of lipase from Burkholderia cepacia dehydrated to form a film. Spectroscopic data show that the additive has little effect on the structure of the protein; however, H/D exchange kinetic and fluorescence anisotropy suggest a more flexible enzyme molecule when in the presence of MePEG. By infrared spectroscopy, we estimated that, after conditioning the films at water activity of 1, the water content in the lipase dehydrated with MePEG is 5.4- and 4.7-fold higher than in the absence of the additive and the additive alone, respectively. Additionally, our infrared data suggest that MePEG acts by hindering intermolecular protein-protein interactions and contributing to increase the accessibility and flexibility of the lipase in the dehydrated solid film. These factors also explain the enhancement of the enzyme catalytic activity (i.e., up to 3.7-fold in neat organic solvent) when in the presence of MePEG. The method and results presented might better address the use of additives for the preparation of e...Continue Reading

References

Nov 21, 1995·Proceedings of the National Academy of Sciences of the United States of America·K Griebenow, A M Klibanov
Apr 1, 1999·Biotechnology and Bioengineering·B A BedellJ S Dordick
Apr 1, 1999·Biotechnology and Bioengineering·F SecundoP L Overbeeke
Aug 18, 1999·Proceedings of the National Academy of Sciences of the United States of America·L Dai, A M Klibanov
Aug 15, 2000·Angewandte Chemie·G Carrea, S Riva
Feb 24, 2001·Nature·A M Klibanov
Feb 28, 2001·Progress in Biophysics and Molecular Biology·A Barth
Mar 20, 2001·Biotechnology and Bioengineering·F SecundoR Morrone
Jun 21, 2001·Journal of Agricultural and Food Chemistry·A M Rakotonirainy, G W Padua
Jan 16, 2002·Biochemistry·Konstantin K TuroverovIrina M Kuznetsova
Sep 27, 2002·Current Opinion in Biotechnology·Moo-Yeal Lee, Jonathan S Dordick
Dec 28, 2002·Journal of Agricultural and Food Chemistry·Yukiko MizutaniTomohiko Mori
Jul 17, 2003·Chemistry : a European Journal·Francesco Secundo, Giacomo Carrea
Jun 23, 2004·European Journal of Biochemistry·Munishwar N Gupta, Ipsita Roy
Jun 30, 2005·Biopolymers·K Liltorp, Y Maréchal

❮ Previous
Next ❯

Citations

Nov 6, 2008·Biotechnology and Bioengineering·Ezio FasoliGabriel L Barletta
Oct 23, 2009·Chembiochem : a European Journal of Chemical Biology·Ricardo J F BrancoJürgen Pleiss
Mar 14, 2013·Chemical Society Reviews·Francesco Secundo

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.