Convenient synthetic approach for tri- and tetraprenylated cyclodipeptides by consecutive enzymatic prenylations

The prenyltransferases EchPT1 and EchPT2 from Aspergillus ruber are responsible for the consecutive prenylations of cyclo-L-Trp-L-Ala, leading to the formation of the triprenylated echinulin as the predominant product. In this study, we demonstrate that EchPT1 also accepts all stereoisomers of cyclo-Trp-Ala and cyclo-Trp-Pro and catalyses regiospecific reverse C2-prenylation at the indole nucleus. EchPT1 products were well accepted by EchPT2 for multiple consecutive prenylations, with conversion yields of 84 to 98% for six of the eight substrates. C2-, C5- and C7-triprenylated derivatives are identified as major enzyme products, with product yields of 40 to 86% in seven cases. High product yields of 25-36%, i.e. approximate 30% of the total enzyme products, were observed for tetraprenylated derivatives in the four reaction mixtures with one D- and one L-configured amino acid residues. To the best of our knowledge, enzymatic preparation of tetraprenylated cyclodipeptides with such high efficacy has not been reported prior to this study.