Engineering in Life Sciences (Jun 2021)

One‐step selective affinity purification and immobilization of His‐tagged enzyme by recyclable magnetic nanoparticles

  • Li‐Jian Zhou,
  • Rui‐Fang Li,
  • Xue‐Yong Li,
  • Ye‐Wang Zhang

DOI
https://doi.org/10.1002/elsc.202000093
Journal volume & issue
Vol. 21, no. 6
pp. 364 – 373

Abstract

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Abstract The NiFe2O4 magnetic nanoparticles (NF‐MNPs) were prepared for one‐step selective affinity purification and immobilization of His‐tagged recombinant glucose dehydrogenase (GluDH). The prepared nanoparticles were characterized by a Fourier‐transform infrared spectrophotometer and microscopy. The immobilization and purification of His‐tagged GluDH on NF‐MNPs were investigated. The optimal immobilization conditions were obtained that mixed cell lysis and carriers in a ratio of 0.13 in pH 8.0 Tris‐HCl buffer at 30℃ and incubated for 2 h. The highest activity recovery and protein bindings were 71.39% and 38.50 μg mg–1 support, respectively. The immobilized GluDH exhibited high thermostability, pH‐stability and it can retain more than 65% of the initial enzyme after 10 cycles for the conversion of glucose to gluconolactone. Comparing with a commercial Ni‐NTA resin, the NF‐MNPs displayed a higher specific affinity with His‐tagged recombinant GluDH.

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