Optimized immobilization of transketolase from E. coli in MgAl-layered double hydroxides.
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 cm(2).
Clermont Université, Université Blaise Pascal, Institut de Chimie de Clermont-Ferrand, ICCF UMR-CNRS 6296, F-63000 Clermont-Ferrand, France.
Nadia
N
Touisni
Franck
F
Charmantray
Virgil
V
Helaine
Claude
C
Forano
Laurence
L
Hecquet
Christine
C
Mousty
D000327:Adsorption; D000536:Aluminum Hydroxide / Q000737:chemistry*; D055162:Biocatalysis; D015374:Biosensing Techniques / Q000379:methods; D011232:Chemical Precipitation; D004338:Drug Combinations; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000378:metabolism; D008276:Magnesium Hydroxide / Q000737:chemistry*; D033362:Powder Diffraction; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties; D014174:Transketolase / Q000378:metabolism*
D004338:Drug Combinations; D004800:Enzymes, Immobilized; D029968:Escherichia coli Proteins; C013591:aluminum hydroxide, magnesium hydroxide, drug combination; D000536:Aluminum Hydroxide; D014174:Transketolase; D008276:Magnesium Hydroxide
Biohybrids; Enzyme immobilization; Layered double hydroxides; Transketolase from E. coli