Abstract
We, report for the first time, the synthesis and kinetics of Ni and Fe3O4 nanobioconjugates obtained by covalent conjugation of nanoparticles to lactate dehydrogenase (LDH). The conjugation reaction was performed using bis(sulfosuccinimidyl) suberate (BS3) as a cross-linking agent. The kinetics of discrete conjugates was studied through conversion of lactate and pyruvate and subsequent reduction of NAD+ to NADH at 340 nm. The enzyme velocities and the Km values designated that LDH-Ni conjugate developed a higher affinity for substrate. The fabrication of LDH-Ni conjugates makes them promising candidates for biotechnology applications. These novel findings pave way to device a convenient nanoparticle-based LDH assay void of any radioactive isotope or use of complicated dyes for invitro detection and management of malignancies such as leukemia, malaria, to determine cytotoxic potential of compounds in food, cosmetics and pharmaceutical industries. Furthermore, the altered comportment of novel conjugates may serve as a guideline in the development of nanobioconjugates and new advances in nanoparticle based diagnosis and therapeutics.
Keywords: Nanobioconjugates, Metal nanoparticles, LDH-Ni conjugate, LDH- Fe3O4 conjugate, Enzyme affinity, Km, Enzyme kinetics, Lactate dehydrogenase, cancer
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