C7-prenylation of tryptophanyl and O-Prenylation of tyrosyl residues in dipeptides by an aspergillus terreus prenytransferase


During our search for novel prenyltransferases, a putative gene ATEG_04218 from Aspergillus terreus raised our attention and was therefore amplified from strain DSM 1958 and expressed in Escherichia coli. Biochemical investigations with the purified recombinant protein and different aromatic substrates in the presence of dimethylallyl diphosphate revealed the acceptance of all the tested tryptophan-containing cyclic dipeptides. Structure elucidation of the main enzyme products by NMR and MS analyses confirmed the attachment of the prenyl moiety to C-7 of the indole ring, proving the identification of a cyclic dipeptide C7-prenyltransferase (CdpC7PT). For some substrates, reversely C3- or N1-prenylated derivatives were identified as minor products. In comparison to the known tryptophan-containing cyclic dipeptide C7-prenyltransferase CTrpPT from Aspergillus oryzae, CdpC7PT showed a much higher substrate flexibility. It also accepted cyclo-L-Tyr-L-Tyr as substrate and catalyzed an O-prenylation at the tyrosyl residue, providing the first example from the dimethylallyltryptophan synthase (DMATS) superfamily with an O-prenyltransferase activity towards dipeptides. Furthermore, products with both C7-prenyl at tryptophanyl and O-prenyl at tyrosyl residue were detected in the reaction mixture of cyclo-L-Trp-L-Tyr. Determination of the kinetic parameters proved that (S)-benzodiazepinedione consisting of a tryptophanyl and an anthranilyl moiety was accepted as the best substrate with a K M value of 204.1 μM and a turnover number of 0.125 s(-1). Cyclo-L-Tyr-L-Tyr was accepted with a K M value of 1,411.3 μM and a turnover number of 0.012 s(-1).


Carsten Wunsch, Hui-Xi Zou, Uwe Linne, Shu-Ming Li


Applied Microbiology and Biotechnology

Year, Volume, Page

2015, 99, 1719



Tag Element Regiochemistry Product Substrate Cofactor Enzyme
PTDBREC00529 C Regular FgaPT2
PTDBREC00530 C Regular CTrpPT
PTDBREC00531 C Reverse CdpC3PT
PTDBREC00532 C Reverse CdpC2PT
PTDBREC00533 C Regular CdpC7PT
PTDBREC00534 C Regular CTrpPT
PTDBREC00535 C Regular CdpC7PT
PTDBREC00536 C Regular CTrpPT
PTDBREC00537 C Regular CdpC7PT
PTDBREC00538 C Regular CTrpPT
PTDBREC00539 C Regular CdpC7PT
PTDBREC00540 C Regular 5-DMATS
PTDBREC00541 C Regular CTrpPT
PTDBREC00542 C Regular CdpC7PT
PTDBREC00543 C Regular CTrpPT
PTDBREC00544 C Regular CdpC7PT
PTDBREC00545 C Regular CTrpPT
PTDBREC00546 C Regular CdpC7PT
PTDBREC00547 C Regular CTrpPT
PTDBREC00548 C Regular 6-DMATS
PTDBREC00549 C Regular 7-DMATS
PTDBREC00550 O Regular CdpC7PT
PTDBREC00551 O Regular CTrpPT
PTDBREC00552 C Reverse AnaPT
PTDBREC00553 C Reverse CdpNPT
PTDBREC00555 C Regular FtmPT1
PTDBREC00554 C Reverse BrePT
PTDBREC00556 N Reverse CTrpPT