A linear pathway for mevalonate production supports growth of Thermococcus kodakarensis
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The sole unifying feature of Archaea is the use of isoprenoid-based glycerol lipid ethers to compose cellular membranes. The branched hydrocarbon tails of archaeal lipids are synthesized via the polymerization of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), but many questions still surround the pathway(s) that result in production of IPP and DMAPP in archaeal species. Isotopic-labeling strategies argue for multiple biological routes for production of mevalonate, but biochemical and bioinformatic studies support only a linear pathway for mevalonate production. Here, we use a combination of genetic and biochemical assays to detail the production of mevalonate in the model archaeon Thermococcus kodakarensis. We demonstrate that a single, linear pathway to mevalonate biosynthesis is essential and that alternative routes of mevalonate production, if present, are not biologically sufficient to support growth in the absence of the classical mevalonate pathway resulting in IPP production from acetyl-CoA. Archaeal species provide an ideal platform for production of high-value isoprenoids in large quantities, and the results obtained provide avenues to further increase the production of mevalonate to drive isoprenoid production in archaeal hosts.
KeywordsMevalonate MVA pathway HMG-CoA reductase Isoprenoid Thermococcus kodakarensis
We thank members of the Santangelo laboratory for assistance with manuscript preparation and editing. These studies were supported by funding from the Department of Energy, Basic Energy Sciences Division, Grant DE-SC0014597 to TJS.
- Bochar DA, Brown JR, Doolittle WF et al (1997) 3-hydroxy-3-methylglutaryl coenzyme A reductase of Sulfolobus solfataricus: DNA sequence, phylogeny, expression in Escherichia coli of the hmgA gene, and purification and kinetic characterization of the gene product. J Bacteriol 179:3632–3638CrossRefGoogle Scholar
- Farkas JA, Picking JW, Santangelo TJ (2013) Genetic techniques for the archaea. Annu Rev Genet 47:539–561. https://doi.org/10.1146/annurev-genet-111212-133225 CrossRefGoogle Scholar
- Rossoni L, Hall SJ, Eastham G et al (2015) The putative mevalonate diphosphate decarboxylase from Picrophilus torridus is in reality a mevalonate-3-Kinase with high potential for bioproduction of isobutene. Appl Environ Microbiol 81:2625–2634. https://doi.org/10.1128/AEM.04033-14 CrossRefGoogle Scholar
- Yamauchi N (2010) The pathway of leucine to mevalonate in halophilic archaea: efficient incorporation of leucine into isoprenoidal lipid with the involvement of isovaleryl-CoA dehydrogenase in Halobacterium salinarum. Biosci Biotechnol Biochem 74:443–446. https://doi.org/10.1271/bbb.90814 CrossRefGoogle Scholar
- Yamauchi N, Tanoue R (2017) Deuterium incorporation experiments from (3 R)- and (3 S)-[3-2H]leucine into characteristic isoprenoidal lipid-core of halophilic archaea suggests the involvement of isovaleryl-CoA dehydrogenase. Biosci Biotechnol Biochem 81:2062–2070. https://doi.org/10.1080/09168451.2017.1373588 CrossRefGoogle Scholar