Determination of Significance:
Saccharifying enzyme, glucoamylase (EC3.2.1.3) also known as γ- amylase, which is an extracellular enzyme that is secreted by a series of microorganisms and has exonuclease activity. The main role of Saccharifying enzyme is to hydrolyze α-1,4 glycosidic bonds in sequence from non-reducing ends on the carbon chains such as starch, dextrin, glycogen, etc., and cut off each glucose unit. For amylopectin, when it encounters a branch point, it can also hydrolyze α-1,6 glycosidic bonds, thereby hydrolyzing all amylopectin to glucose. It is one of the important industrial enzyme preparations in China, and is widely used in alcohol, liquor, antibiotics, amino acids, organic acids, glycerol, starch sugar and other industries.
Measurement Principle:
Soluble starch could be hydrolyzed into glucose by saccharifying enzyme. 3, 5-Dinitrosalicylic acid is reduced to brown-red amino compound by co-heating with glucose under alkaline condition. The brown-red amino compound has a maximum absorption peak at 540 nm and the absorbance ratio is in direct proportion to the contents of glucose. The saccharifying enzyme activity is quantified by measuring the color development at 540 nm.
Self Provided:
Microplate reader/spectrophotometer, water bath, balance, desk centrifuge, micro glass cuvette/96 well flat-bottom plate, adjustable transferpettor, mortar/homogenizer/ultrasonic crusher, ice and distilled water.
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