caa a22 4500 605503834 CHVBK 20210128100612.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00253-014-5999-6 doi (NATIONALLICENCE)springer-10.1007/s00253-014-5999-6 C7 -prenylation of tryptophanyl and O -prenylation of tyrosyl residues in dipeptides by an Aspergillus terreus prenyltransferase [Elektronische Daten] [Carsten Wunsch, Hui-Xi Zou, Uwe Linne, Shu-Ming Li] 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 204.1μM and a turnover number of 0.125s−1. Cyclo-l-Tyr-l-Tyr was accepted with a K M value of 1,411.3μM and a turnover number of 0.012s−1. Springer-Verlag Berlin Heidelberg, 2014 Aspergillus terreus nationallicence Cyclic dipeptide nationallicence DMATS superfamily nationallicence Prenyltransferase nationallicence Tryptophan C7 -prenylation nationallicence Tyrosine O -prenylation nationallicence Wunsch Carsten Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Deutschhausstraße 17A, 35037, Marburg, Germany aut Zou Hui-Xi Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Deutschhausstraße 17A, 35037, Marburg, Germany aut Linne Uwe Fachbereich Chemie, Philipps-Universität Marburg, Hans-Meerwein-Straße, 35032, Marburg, Germany aut Li Shu-Ming Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Deutschhausstraße 17A, 35037, Marburg, Germany aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1719-1730 0175-7598 99:4<1719 2015 99 253 https://doi.org/10.1007/s00253-014-5999-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-5999-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wunsch Carsten Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Deutschhausstraße 17A, 35037, Marburg, Germany aut NATIONALLICENCE 700 1- Zou Hui-Xi Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Deutschhausstraße 17A, 35037, Marburg, Germany aut NATIONALLICENCE 700 1- Linne Uwe Fachbereich Chemie, Philipps-Universität Marburg, Hans-Meerwein-Straße, 35032, Marburg, Germany aut NATIONALLICENCE 700 1- Li Shu-Ming Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Deutschhausstraße 17A, 35037, Marburg, Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1719-1730 0175-7598 99:4<1719 2015 99 253