Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Robert-Koch-Straße 4, 35037, Marburg, Germany.
Appl Microbiol Biotechnol. 2018 Mar;102(6):2671-2681. doi: 10.1007/s00253-018-8761-7. Epub 2018 Jan 25.
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.
红曲霉中的 prenyltransferases EchPT1 和 EchPT2 负责连续 prenylation 的环-L-Trp-L-Ala，导致 triprenylated echinulin 的形成作为主要产物。在这项研究中，我们证明 EchPT1 也接受所有立体异构体的环-Trp-Ala 和环-Trp-Pro，并催化在吲哚核的区域特异性反向 C2-prenylation。EchPT1 的产物被 EchPT2 很好地接受进行多次连续 prenylation，对于八个底物中的六个，转化率为 84%至 98%。C2-、C5-和 C7-三 prenylated 衍生物被鉴定为主要的酶产物，在七种情况下的产率为 40%至 86%。在四个反应混合物中，对于含有一个 D-和一个 L-构型氨基酸残基的四 prenylated 衍生物，观察到高产物产率为 25-36%，即大约占总酶产物的 30%。据我们所知，在此之前，这种高效的四 prenylated 环二肽的酶制备尚未有报道。
