Determination of Total Available Carbohydrates in Forage Legume Roots by Extraction with Takadiastase, Amyloglucosidase, or Sulfuric Acid1
- L. J. Greub and
- W. F. Wedin2
Takadiastase (“Clarase 900”), amyloglucosidase (“Diazyme 160”), and a combination of the two enzymes were compared with and without acid hydrolysis of the filtrates for the ability to extract sucrose and starch and convert them to reducing sugars in the determination of total available carbohydrates (TAC). Sucrose, soluble starch, and alfalfa root samples varying in TAG content and ratio of sucrose to starch were used as substrates. Comparisons also were made between TAC values obtained by the enzyme combination and by O.2N H2SO4. Clarase 900 recovered significantly more sucrose but significantly less starch than did Diazyme 160 or the enzyme combination. A mild acid hydrolysis subsequent to the enzyme stage enhanced sucrose recovery but not starch recovery.
When used on the root material without acid hydrolysis, the Clarase 900 and the enzyme combination gave similar TAC values. Diazyme 160 values were significantly lower than the other two treatments. Acid hydrolysis of the enzyme filtrate increased the TAC values of most of the root samples.
The enzyme combination method gave TAC values up to three-fold higher than the 0.2N H2SO4 method when legume root samples were high in starch content, but the acid method gave higher values when TAC were low.Please view the pdf by using the Full Text (PDF) link under 'View' to the left.
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