Synthesis of high molecular weight polyphosphate with a partially purified enzyme from Salmonella.

Polyphosphate (polyp) kinase was prepared from EDTA-lysozyme spheroplasts of a Salmonella minnesota rough mutant. The enzyme, purified about 8o-fold, was free of ATPase and polyp-degrading activity and contained neither nucleic acids nor polyp which might act as primer. Using this enzyme, 32P-labelled polyp was prepared from [?P]ATP. The polyp was identified by the action of degrading enzymes, phenol extraction, adsorption characteristics on charcoal and hydrolysis kinetics. Sedimentation data in sucrose gradients (S = I 1.5) and gel filtration on agarose indicated a molecular weight in the range 300,000 to 400,000 daltons.