Branching enzyme assay: Selective quantitation of the α1,6-linked glucosyl residues involved in the branching points

Methods previously described for glycogen or amylopectin branching enzymatic activity are insufficiently sensitive and not quantitative. A new, more sensitive, specific, and quantitative one was developed. It is based upon the quantitation of the glucose residues joined by α1,6 bonds introduced by v...

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Autores principales: Krisman, C.R., Tolmasky, D.S., Raffo, S.
Formato: JOUR
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Acceso en línea:http://hdl.handle.net/20.500.12110/paper_00032697_v147_n2_p491_Krisman
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Sumario:Methods previously described for glycogen or amylopectin branching enzymatic activity are insufficiently sensitive and not quantitative. A new, more sensitive, specific, and quantitative one was developed. It is based upon the quantitation of the glucose residues joined by α1,6 bonds introduced by varying amounts of branching enzyme. The procedure involved the synthesis of a polysaccharide from Glc-1-P and phosphorylase in the presence of the sample to be tested. The branched polysaccharide was then purified and the glucoses involved in the branching points were quantitated after degradation with phosphorylase and debranching enzymes. This method appeared to be useful, not only in enzymatic activity determinations but also in the study of the structure of α-d-glucans when combined with those of total polysaccharide quantitation, such as iodine and phenol-sulfuric acid. © 1985.