Open AccessOne‐step selective affinity purification and immobilization of His‐tagged enzyme by recyclable magnetic nanoparticles
Author(s) -
Zhou LiJian,
Li RuiFang,
Li XueYong,
Zhang YeWang
Publication year - 2021
Publication title -
engineering in life sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.547
H-Index - 57
eISSN - 1618-2863
pISSN - 1618-0240
DOI - 10.1002/elsc.202000093
Subject(s) - thermostability , chemistry , magnetic nanoparticles , immobilized enzyme , lysis , nuclear chemistry , recombinant dna , nanoparticle , tris , chromatography , enzyme , affinity chromatography , fourier transform infrared spectroscopy , biochemistry , chemical engineering , nanotechnology , materials science , engineering , gene
The NiFe 2 O 4 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.