期刊
TALANTA
卷 258, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.talanta.2023.124406
关键词
Enzyme immobilization; Biomimetic magnetic nanoparticles; Stability; Reusability
The immobilization of enzymes on magnetic nanoparticles has attracted interest in detecting contaminants in water samples due to their magnetic control, concentration, and reuse capabilities. In this study, nanoassemblies composed of inorganic or biomimetic magnetic nanoparticles were developed for the detection of trace amounts of organophosphate pesticides and antibiotics in water. Enzyme immobilization was achieved through electrostatic interaction or covalent bonds, with covalent bonding yielding better results. The optimized nanoassemblies demonstrated the ability to detect pollutants at low concentrations and provided enhanced stability and reuse of the enzymes.
The use of enzymes immobilized on magnetic nanoparticles to detect contaminants in aqueous samples has gained interest, since it allows the magnetic control, concentration and reuse of the enzymes. In this work, the detection of trace amounts of organophosphate pesticides (chlorpyrifos) and antibiotics (penicillin G) in water was attained by developing a nanoassembly formed by either inorganic or biomimetic magnetic nanoparticles used as substrates to immobilize acetylcholinesterase (AChE) and beta-lactamase (BL). Other than the substrate, the optimization of the nanoassembly was done by testing enzyme immobilization both through electrostatic interaction (also reinforced with glutaraldehyde) and covalent bonds (by carbodiimide chemistry). Temperature (25 degrees C), ionic strength (150 mM NaCl) and pH (7) were set to ensure enzymatic stability and to allow both the nanoparticles and the enzymes to present ionic charges that would allow electrostatic interaction. Under these conditions, the enzyme load on the nanoparticles was-0.1 mg enzyme per mg nanoparticles, and the preserved activity after immobilization was 50-60% of the specific activity of the free enzyme, being covalent bonding the one which yielded better results. Covalent nanoassemblies could detect trace concentrations of pollutants down to 1.43 nM chlorpyrifos and 0.28 nM penicillin G. They even permitted the quantification of 14.3 mu M chlorpyrifos and 2.8 mu M penicillin G. Also, immobilization conferred higher stability to AChE (-94% activity after 20 days storage at 4 degrees C) and allowed to reuse the BL up to 12 cycles.
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