Changes in the activity of enzymes involved in purine “salvage” and nucleic acid degradation during the growth of Catharanthus roseus cells in suspension culture

Abstract

Changes during growth in the activity of several enzymes involved in purine “salvage”, adenine phosphoribosyltransferase (EC 2.4.2.7), guanine phosphoribosyl‐transferase (EC 2.4.2.8), hypoxanthine phosphoribosyltransferase (EC 2.4.2.8) and adenosine kinase (EC 2.7.1.20), the enzymes which catalyze the conversion of nucleoside monophosphate to triphosphate, nucleoside monophosphate kinase (EC 2.7.4.4) and nucleoside diphosphate kinase (EC 2.7.4.6), and several degradation enzymes, deoxyribonucleae(s), ribonuclease(s). phosphatase(s), nucleosidase (EC 3.2.2.1), 3′‐nucleotidase (EC 3.1.3.6) and 5′‐nucleotidase (EC 3.1.3.5) were examined in cells of Catharanthus roseus (L.) G. Don cultured in suspension. In addition, the incorporation of [8‐14C] adenine, [8‐14C] adenine, [8‐14C]hypoxanthine. [8‐14C] adenosine and [8‐14C]inosine into nucleotides and nucleic acids was also determined using intact cells.The activities of all purine “salvage” enzymes examined and those of nucleoside monophosphate and diphosphate kinases increased rapidly during the lag phase and decreased during the following cell division and cell expansion phases. The rate of incorporation of adenine, guanine, hypoxanthine, and adenosine into nucleotides and nucleic acids was higher in the lag phase cells than during the following three phases. The highest rate of [8‐14C]inosine incorporation was observed in the stationary phase cells. The activity of all degradation enzymes examined decreased when the stationary phase cells were transferred to a new medium.These results indicated that the increased activity of purine “salvage” enzymes observed in the lag phase cells may contribute to an active purine “salvage” which is required to initiate a subsequent cell division.