Characterization of a Feedback-Resistant Mevalonate Kinase from the Archaeon Methanosarcina mazei

Abstract

The mevalonate pathway is utilized for the biosynthesis of isoprenoids in many bacterial, eukaryotic, and archaeal organisms. Based on previous reports of its feedback inhibition, mevalonate kinase (MVK) may play an important regulatory role in the biosynthesis of mevalonate pathway-derived compounds. Here we report the purification, kinetic characterization, and inhibition analysis of the MVK from the archaeon Methanosarcina mazei. The inhibition of the M. mazei MVK by the following metabolites derived from the mevalonate pathway was explored: dimethylallyl diphosphate (DMAPP), geranyl pyrophosphate (GPP), farnesyl pyrophosphate (FPP), isopentenyl monophosphate (IP), and diphosphomevalonate. M. mazei MVK was not inhibited by DMAPP, GPP, FPP, diphosphomevalonate, or IP, a proposed intermediate in an alternative isoprenoid pathway present in archaea. Our findings suggest that the M. mazei MVK represents a distinct class of mevalonate kinases that can be differentiated from previously characterized MVKs based on its inhibition profile.

An in Vitro Biosynthesis of Sesquiterpenes Starting from Acetic Acid

“…Here, MVKs from the thermophilic archaeon M. jannaschii [10] and the archaeon M. mazei [11] were compared. The enzyme from M. jannaschii is described to be feedback inhibited by GPP and FPP, whereas the latter enzyme is described to be feedback resistant . In our experiments, both enzymes yielded the same results.…”

(R)-Mevalonate 3-Phosphate Is an Intermediate of the Mevalonate Pathway in Thermoplasma acidophilum

“…Therefore, it remains unclear how IP is synthesized in most archaea. Although they possess an MVK homolog gene, and MVK activity has been identified in a few cases (21,22), it is also unclear which of the two modified MVA pathways, i.e. the Roseiflexus/Haloferax-type pathway via MVA-5-P and the Thermoplasma-type pathway via MVA-3-P, is actually used by them.…”

Lipid sugar carriers at the extremes: The phosphodolichols Archaea use in N-glycosylation

“…volcanii HMG-CoA reductase is the first such enzyme that is not sensitive to feedback inhibition by its substrate, acetoacetyl-CoA [68]. Likewise, known feedback inhibitors of mevalonate-5-kinases do not inhibit the Methanosarcina mazei enzyme, making it the sole member of a third class of mevalonate kinases defined on the basis of inhibition profile [69]. …”