RT Journal Article SR Electronic(1) A1 Mundt, Kathrin A1 Li, Shu-MingYR 2013 T1 CdpC2PT, a reverse prenyltransferase from Neosartorya fischeri with a distinct substrate preference from known C2-prenyltransferases JF Microbiology, VO 159 IS Pt_10 SP 2169 OP 2179 DO https://doi.org/10.1099/mic.0.069542-0 PB Microbiology Society, SN 1465-2080, AB A putative prenyltransferase gene, NFIA_043650, was amplified from Neosartorya fischeri NRRL 181 and cloned into the expression vector pQE60. The deduced polypeptide consisting of 445 amino acids with a molecular mass of 51 kDa was overproduced in Escherichia coli and purified as His6-tagged protein to near homogeneity. The purified soluble protein was subsequently assayed with potential aromatic substrates in the presence of dimethylallyl diphosphate. HPLC analysis of the reaction mixtures revealed acceptance of all tested tryptophan-containing cyclic dipeptides. Isolation and structural elucidation of enzyme products of five selected substrates indicated a reverse C2-prenylation on the indole nucleus, proving the enzyme to be a cyclic dipeptide C2-prenyltransferase (CdpC2PT). Differing significantly from two known brevianamide F reverse C2-prenyltransferases NotF and BrePT which use cyclo-l-Trp-l-Pro as their preferred substrate, CdpC2PT showed a clear substrate preference for (S)-benzodiazepinedinone and cyclo-l-Trp-l-Trp with K M values of 84.1 and 165.2 µM and turnover numbers at 0.63 and 0.30 s−1, respectively. A possible role of CdpC2PT in the biosynthesis of fellutanines is discussed., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.069542-0