Determination of Significance:
L-malic acid is an intermediate product of the tricarboxylic acid cycle and an important part of the malate-aspartic acid shuttle. Malate-aspartic acid shuttle is required for the reductive equivalent transmitochondrial membrane transport process of oxidative phosphorylation. In lower organisms, malic acid is converted to lactic acid in the process of malolactic fermentation, which also produces CO2. Malic acid is commonly used as an additive in the food and pharmaceutical industries, and quantitative analysis of malic acid is also vital in the production of beer, wine, cheese and fruit.
Measurement Principle:
Malate dehydrogenase catalyzes malic acid and NAD to form oxaloacetic acid, NADH and NH4+. Under 1-mPMS, WST-1 reacts with NADH to produce water-soluble Formazan. The maximum absorption peak at 450nm can be used to calculate malic acid content.
Self Provided:
Spectrophotometer/Microplate reader, mortar/homogenizer/cell ultrasonic crusher, centrifuge, constant temperature foster box/water-bath, centrifuge, micro quartz cuvette/96 well flat-bottom plate, ice and distilled water.
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