In vivo characterization of the activities of novel cyclodipeptide oxidases: new tools for increasing chemical diversity of bioproduced 2,5-diketopiperazines in Escherichia coli

摘要: Cyclodipeptide oxidases (CDOs) are enzymes involved in the biosynthesis of 2,5-diketopiperazines, a class naturally occurring compounds with large range pharmaceutical activities. CDOs belong to cyclodipeptide synthase (CDPS)-dependent pathways, which they play an early role chemical diversification cyclodipeptides by introducing Cα-Cβ dehydrogenations. Although activities more than 100 CDPSs have been determined, only few characterized. Furthermore, assessment CDO on chemically-synthesized has shown these be relatively promiscuous, making them interesting tools for diversification. The purpose this study is provide first completely microbial toolkit efficient bioproduction variety dehydrogenated 2,5-diketopiperazines. We mined genomes encoded biosynthetic gene clusters CDPS-dependent pathways and selected several characterization. co-expressed each their associated CDPS pathway using Escherichia coli as chassis showed that derivatives were produced culture supernatants. determined biological six novel solving structures biologically cyclodipeptides. Then, we assessed plus two previously characterized combinatorial engineering experiments E. coli. eight 18 diversity synthesize. detected 50 best CDPS/CDO combinations optimize production 23. Our establishes usefulness It constitutes step toward complex diverse