Probing the Substrate Promiscuity of Isopentenyl Phosphate Kinase as a Platform for Hemiterpene Analogue Production1. (22nd July 2019)

• Record Type: Journal Article
• Authors: Lund, Sean; Courtney, Taylor; Williams, Gavin J.
• Is Part Of: Chembiochem. Volume 20:Number 17(2019)
• Page Start: 2217
• Page End: 2221
• Publication Date: 2019-07-22
• Subjects: hemiterpenes -- kinases -- phosphorylation -- synthetic biology -- terpenoids; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201900135
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: 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. Abstract : Expanding terpenoid structural diversity : Isopentenyl phosphate kinase is a key component of an artificial pathway for hemiterpene production. Its specificity was probed with a panel of unnatural analogues, revealing promiscuity towards a variety of alkenyl, alkynyl, and aromatic monophosphates. This sets the stage for probing the ability of artificial hemiterpene pathways to provide precursors for isoprenoid biosynthesis.