Abstract
Publisher Summary Escherichia colt strain K12 is used for the preparation of the cAMP phosphodiesterase. Conditions of growth, e.g., the use of either mineral salts-based or complex media, do not appear to affect the specific cyclic AMP phosphodiesterase activity significantly. Cells may be stored frozen at -20°C or below for prolonged periods of time. E. colt strain Crookes (American Type Culture Collection 8739) when grown on a mineral salts medium is devoid of detectable cAMP phosphodiesterase activity and serves as source of the activator. The bacteria of both strains K12 and Crookes are extracted by any one of three equally effective methods. Extraction and all following operations are carried out at 0–4°C. The bacteria are suspended in a volume (ml) of buffer A equal to their wet weight in grams. They are then disrupted either by passage through a French press at 5000 psi, or by treatment in a Raytheon 10-kcycles/sec magnetostrictive oscillator for two consecutive 15-minute periods, or by grinding with an approximately equal weight of levigated alumina.
Published Version
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