The requirement for a primer in the in vitro synthesis of polysaccharide by sweet-corn (1 → 4)-α-d-glucan syntrase

A mixture of (1 → 4)-α-D-glucan synthases was partially purified from sweet corn. The synthesis of polysaccharide from ADP-D-glucose by the enzyme preparation was dependent on added carbohydrate primer in solutions of low ionic strength, but displayed the phenomenon of being apparently primer-independent at high ionic strength in citrate buffer. This phenomenon was further investigated; treatment of the enzyme preparation with immobilized amylases led to the abolition of the apparently unprimed synthesis. The amylase-treated preparation then showed a normal dependence on (1 → 4)-α-D-glucan primer, branched primers being the most effective. The affinity of the enzyme for a branched primer appeared to be enhanced in the presence of citrate. The polysaccharide product of the unprimed reaction was glycogen-like, having an average chain-length of 14. These studies suggest that the phenomenon of unprimed synthesis in "high salt" is explicable in terms of an enhanced affinity of the enzyme for traces of primer in the enzyme preparation, and not to a "de novo" synthesis of polysaccharide that occurs in the absence of a primer.