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Determination of Significance：
Acetyl coenzyme a is an important intermediate metabolite in the process of energy metabolism, which is widely found in animals, plants, microbes and cultured cells. Three major nutrients (sugar, fat and protein) converge through acetyl coenzyme a to form a common metabolic pathway - tricarboxylic acid cycle and oxidative phosphorylation. Through this pathway, they are completely oxidized to produce carbon dioxide and water, release energy for ATP synthesis. Acetyl coenzyme a is the precursor for synthesis of bioactive substances such as fatty acids, ketones, cholesterol and their derivatives.
Measurement Principle：
Malate Dehydrogenase (MDH) catalyzes NAD+ and malate to generate NADH and oxaloacetate. Citrate Synthase (CS) catalyzes oxaloacetate and acetyl coenzyme a to generate Citrate and Coenzyme A. Because of the coupling reaction of MDH and CS, acetyl coenzyme a content is proportional to NADH production rate. Acetyl coenzyme a could be calculated by changes of light absorption at 340nm.
Self Provided：
Ultraviolet spectrophotometer/Microplate reader, constant temperature foster box/water-bath, centrifuge, transferpettor, micro quartz cuvette/96 well UV plate, mortar/homogenizer, ice and distilled water.
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