Starch Branching Enzyme (SBE) catalyzes the conversion of Amylose to Amylopectin. SBE can reduce the content of amylose and thus reduce the optical absorption value of starchiodine complex at 660 nm. The percentage of absorbance decline in a certain time can reflect the activity of SBE.
Starch Debranching Enzyme (DBE) can specifically split the α-1, 6-glucoside bonds of amylopectin. The resulting reducing sugars were determined by 3, 5-dinitrosalicylic acid method, and their light absorption values were measured at 540 nm. Then the activity of starch debranching enzyme was calculated indirectly.
Starch Phosphorylase (SP) breaks down the substrate to produce α-D-glucose-1 phosphate, which is converted to α-D-glucose-6 phosphate by the enzyme. It also reduces NAD+/NADP+ with the catalysis of dehydrogenase to produce NADH/NADPH, and the absorbance value at 340 nm increases.
SDH catalyzes the dehydrogenation of succinate to fumarate, with electron transport materials transferring electrons to 2, 6-dichlorophenol indiophenol (DCPIP). Then, the reduced DCPIP is reduced to the oxidized DCPIP, which has a characteristic absorption peak at 600 nm. Therefore, the activity of SDH can be quantified by measure the change OD value at 600 nm.
Succinic acid, also known as succinic acid, is widely present in all plant and animal tissues, was first extracted from amber, is an intermediate in the citric acid cycle, and plays an important role in the production of energy in cells. Succinate, or succinate, is widely used in the agricultural, food and pharmaceutical industries due to its low toxicity. Succinic acid is catalyzed by enzyme reagent to form colored substance with color developer. The maximum absorption is at 555 nm. The content of succinic acid in samples can be calculated by measuring OD value and standard curve at 555 nm.
Succinyl-CoA synthetase (SCS) is a key enzyme in the citric acid cycle and an important metabolic intermediate in the biosynthesis of porphyrins, hemes, and ketones. SCS converts succinyl-coenzyme A (CoA-SH) to succinic acid. The reaction between CoA-SH and pyruvate produces chromogenic substances with a characteristic absorption peak at 450 nm. The activity of SCS can be calculated by measuring the change of absorbance value at 450 nm.
Sucrase catalyzes its substrate (sucrose) to produce glucose, which produces hydrogen peroxide under the action of glucose oxidase. Hydrogen peroxide reacts with chromogenic agent to produce red substance, which has a strong absorption peak at 505 nm. In a certain concentration range, It's absorbance is proportional to glucose concentration. Therefore, the activity of sucrase can be calculated by measuring the OD value at 505 nm.
Sucrose in plant tissue is hydrolyzed to glucose and fructose in boiling water bath under acidic conditions. 5-hydroxymethyl furfural was synthesized from fructose under acid condition and measure the ultraviolet absorption of 5-hydroxymethyl furfural. Glucose must be dissimilated into ketose structure and reduced to obtain 5-hydroxymethylfurfural, but the rate of isomerization of glucose to ketose is very slow. Therefore, the ultraviolet absorption of glucose is much smaller than fructose.
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