Optimized immobilization of transketolase from E. coli in MgAl-layered double hydroxides

• Published in: Colloids and surfaces, B, Biointerfaces B, Biointerfaces, 2013-12, Vol.112, p.452-459
• Main Authors: Touisni, Nadia, Charmantray, Franck, Helaine, Virgil, Forano, Claude, Hecquet, Laurence, Mousty, Christine
• Format: Article
• Language: English
• Subjects: Adsorption; Aluminum Hydroxide - chemistry; Biocatalysis; Biohybrids; Biosensing Techniques - methods; Chemical Precipitation; colloids; Coprecipitation; Deposition; Diffraction; Drug Combinations; enzyme activity; Enzyme immobilization; Enzyme Stability; Enzymes, Immobilized - metabolism; Escherichia coli; Escherichia coli - enzymology; Escherichia coli Proteins - metabolism; Fourier transform infrared spectroscopy; Hydroxides; Immobilization; Layered double hydroxides; Magnesium Hydroxide - chemistry; Polished; Powder Diffraction; Spectroscopy, Fourier Transform Infrared; Surface chemistry; Surface Properties; transketolase; Transketolase - metabolism; Transketolase from E. coli; X-ray diffraction
• ISSN: 0927-7765; 1873-4367
• DOI: 10.1016/j.colsurfb.2013.07.023

Immobilization of TK from Escherichia coli (TKec) on MgAl-NO3 layered double hydroxides (LDH) was carried out by two processes: adsorption and coprecipitation. As a comparison, the adsorption method was realized either at pH 7.5 in buffered solutions (MOPS and Gly-Gly) or in pure water. For the coprecipitation method, the formation of the inorganic LDH support was realized directly in the presence of TKec solubilized in Gly-Gly. The prepared biohybrids, called respectively TKec@LDHads and TKec@LDHcop, were characterized by powder X-ray diffraction, FTIR spectroscopy in comparison with TKec free reference products, i.e. MgAl-NO3, MgAl-Gly-Gly. The enzymatic activities of the various TKec@LDH biohybrids as well as their stabilities over time were investigated by UV–vis assay. A maximum of activity (12 U/mg of solid) was reached for TKec@MgAl-Gly-Gly biohybrid prepared by coprecipitation. Finally, thin films were prepared through a one-step deposition on a polished support. The enzymatic activity of the resulting TKec@MgAl-Gly-Glycop film was tested over four recycling processes with a reproducible activity of 2.7 U/mg cm2. [Display omitted] •Immobilization of TK from E. coli (TKec) on layered double hydroxides (LDH).•The formation of the inorganic LDH support is realized directly in the presence of TKec solubilized in Gly-Gly.•The immobilized TKec can be reused several times without any loss of activity.