An experimental strategy towards optimising directed biosynthesis of communesin analogues by Penicillium marinum in submerged fermentation

It was previously demonstrated that a fungus producing communesin alkaloids, subsequently identified as Penicillium marinum, could also accept 6-fluoro analogues of tryptophan or tryptamine to form mono-fluoro-communesin analogues in addition to communesins. A strategy to increase the relative yield of analogues by mutation to impair decarboxylation of tryptophan has been studied. Four mutants with much reduced activity of tryptophan decarboxylase, and other phenotypic change, were selected from 1500 colonies from spores that survived a 99 % kill treatment with N-methyl N-nitro N-nitrosoguanidine. TIC assessment of cell-associated products from standard submerged fermentations showed that one non-sporing mutant apparently produced little or no communesins, but productivity was restored when grown in a medium supplemented with glutamine. However, more sensitive mass spectrometric analysis detected both communesins A and B in mycelium grown on a rich, yeast extract-sucrose agar, showing that deletion of communesin biosynthesis was not absolute. It was concluded that mutagenesis had generally achieved its objective, but that new literature on a putative role of aurantioclavine in communesin biosynthesis presented an additional challenge to integrate the prenylation of tryptophan before its decarboxylation, which is a characteristic of ergot alkaloid biosynthesis. (C) 2007 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.