Biochemistry
Pooyan Pedram; Mohammad Fazilati; Marzieh Rashidipour; Habibollah Nazem
Abstract
Lactate dehydrogenase (LDH) is a key enzyme in cellular metabolism found in all animals. It plays a crucial role in converting pyruvate to lactate and vice versa. LDH is present in a wide range of tissues and cells in the animal body. In recent decades, nanoparticles have been utilized due to their unique ...
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Lactate dehydrogenase (LDH) is a key enzyme in cellular metabolism found in all animals. It plays a crucial role in converting pyruvate to lactate and vice versa. LDH is present in a wide range of tissues and cells in the animal body. In recent decades, nanoparticles have been utilized due to their unique properties for designing optical and electronic sensors. This research presents a novel colorimetric method: silver nanoparticles synthesized using chrysanthemum aqueous extract are employed for direct detection of lactate dehydrogenase activity. Initially, chrysanthemums were collected from greenhouses in Mahallat County under the supervision of experts. After separation and powder preparation of the flower part of the plant, chrysanthemum aqueous extract was prepared. Subsequently, the synthesis of silver nanoparticles using aqueous extract and addition of silver nitrate solution was investigated by optimizing appropriate conditions. In the next step, two vials were prepared, each containing a reaction mixture comprising Tris-HCl, MgCl2, and NADH. Additionally, one vial contained LDH. Silver nanoparticles and sodium borohydride were then added to the vials. The enzyme can convert NAD+ to NADH. The detection mechanism of lactate dehydrogenase enzyme is based on the aggregation of silver nanoparticles, which leads to an increase in their size and consequently a color change. Thus, the presence or absence of the enzyme can be easily distinguished with the naked eye in a single step. In the presence of the enzyme, the color of the solution used in the study was yellow, while in the absence of the enzyme, the color was grayish. Consequently, lactate dehydrogenase enzyme can be identified with high sensitivity.
