Abstract
Aromatic prenyltransferases are known for their extensive promiscuity toward aromatic acceptor substrates and their ability to form various carbon-carbon and carbon-heteroatom bonds. Of particular interest among the prenyltransferases is NphB, whose ability to geranylate cannabinoid precursors has been utilized in several in vivo and in vitro systems. It has therefore been established that prenyltransferases can be utilized as biocatalysts for the generation of useful compounds. However, recent observations of non-native alkyl-donor promiscuity among prenyltransferases indicate the role of NphB in biocatalysis could be expanded beyond geranylation reactions. Therefore, the goal of this study was to elucidate the donor promiscuity of NphB using different acceptor substrates. Herein, we report distinct donor profiles between NphB-catalyzed reactions involving the known substrate 1,6-dihydroxynaphthalene and an FDA-approved drug molecule sulfabenzamide. Furthermore, we report the first instance of regiospecific, NphB-catalyzed N-alkylation of sulfabenzamide using a library of non-native alkyl-donors, indicating the biocatalytic potential of NphB as a late-stage diversification tool.Key Points• NphB can utilize the antibacterial drug sulfabenzamide as an acceptor.• The donor profile of NphB changes dramatically with the choice of acceptor.• NphB performs a previously unknown regiospecific N-alkylation on sulfabenzamide.• Prenyltransferases like NphB can be utilized as drug-alkylating biocatalysts.
Highlights
Aromatic prenyltransferases (PTs) comprise a large class of enzymes catalyzing the transfer of a prenyl group from a prenyl-pyrophosphate donor onto an aromatic acceptorBryce P
The donor profiles of 1,6-DHN and sulfabenzamide in reactions catalyzed by NphB showed highly distinct trends, with 1,6-DHN favoring longer aliphatic moieties (~ 8–9 unbranched atoms) and sulfabenzamide showing a preference for shorter ones (~ 5–6 unbranched atoms)
To explain the observed trends, sulfabenzamide was docked in the active site of the NphB crystal structure bound to geranylthiolopyrophosphate (GSPP) and 1,6-DHN (PDB ID: 1ZB6; Kuzuyama et al 2005; Fig. 6a)
Summary
Aromatic prenyltransferases (PTs) comprise a large class of enzymes catalyzing the transfer of a prenyl group from a prenyl-pyrophosphate donor onto an aromatic acceptorBryce P. Aromatic prenyltransferases (PTs) comprise a large class of enzymes catalyzing the transfer of a prenyl group from a prenyl-pyrophosphate donor onto an aromatic acceptor. (Awakawa and Abe 2019; Saleh et al 2009). This reaction scheme, delivers a high level of chemical diversity among the products of PT reactions arising from a combination of factors (Kumano et al 2008; Kuzuyama et al 2005; Shindo et al 2011; Tanner 2015; Tello et al 2008). The accepted prenyl groups can vary in size (5-carbon dimethyallyl, 10-carbon geranyl, 15-carbon farnesyl, 20carbon geranylgeranyl) between PTs, while the attachment can occur from either C1′ or C3′ of the prenyl donors onto various positions of the aromatic acceptors, resulting in normal or reverse prenylation, respectively (Giessen and Marahiel 2015; Winkelblech et al 2015a). PTs have been shown to catalyze the formation of C–C (ArayaCloutier et al 2017; Bandari et al 2019; Elshahawi et al 2017; Fan et al 2015b; Haagen et al 2007; Winkelblech et al 2015b), C–O (Bandari et al 2017; Haagen et al 2007; Appl Microbiol Biotechnol (2020) 104:4383–4395
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