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Probing the Substrate Promiscuity of Isopentenyl Phosphate Kinase as a Platform for Hemiterpene Analogue Production
Author(s) -
Lund Sean,
Courtney Taylor,
Williams Gavin J.
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201900135
Subject(s) - terpenoid , kinase , enzyme , biochemistry , biosynthesis , chemistry , metabolic engineering , thermoplasma acidophilum , mevalonate pathway , stereochemistry , biology
Isoprenoids are a large class of natural products with wide‐ranging applications. Synthetic biology approaches to the manufacture of isoprenoids and their new‐to‐nature derivatives are limited due to the provision in nature of just two hemiterpene building blocks for isoprenoid biosynthesis. To address this limitation, artificial chemo‐enzymatic pathways such as the alcohol‐dependent hemiterpene (ADH) pathway serve to leverage consecutive kinases to convert exogenous alcohols into pyrophosphates that could be coupled to downstream isoprenoid biosynthesis. To be successful, each kinase in this pathway should be permissive of a broad range of substrates. For the first time, we have probed the promiscuity of the second enzyme in the ADH pathway—isopentenyl phosphate kinase from Thermoplasma acidophilum —towards a broad range of acceptor monophosphates. Subsequently, we evaluate the suitability of this enzyme to provide unnatural pyrophosphates and provide a critical first step in characterizing the rate‐limiting steps in the artificial ADH pathway.