Ammonium repression of antibiotic and intracellular proteinase production in Penicillium urticae

In this study the addition of ammonium ions (5-30 mM) to Penicillium urticae shake-flask cultures before, during and after the onset of polyketide biosynthesis was examined in a time-dependent manner for its repressive effect on metabolites and a marker enzyme of the patulin pathway and on the intracellular proteinases that also appear during the non-growth or idiophase. A study of the effect of ammonium ion addition, showed that both secondary enzyme and proteinase appearance were maximally delayed if the addition was made before the normal 7 h period of derepression/induction. If added during this period the effect of ammonium ions was progressively less. A reduction in the extracellular ammonium ion concentration from 30 to 4 mM appeared to be required to initiate the derepression/induction process. Adding ammonium ions during the appearance of secondary enzymes caused a rapid decrease in specific activity, about 67% for the patulin pathway enzyme and 12% for proteinase. Nitrogen repression exerts a much stronger effect on the expression of polyketide genes as opposed to proteinase genes. Both patulin pathway enzymes and proteinases are subjected to proteolysis, but the proteinases retain much of their activity, whereas the polyketide biosynthetic enzymes do not.