Molecular cloning, characterization, and functional analysis of a gene encoding 3-hydroxy-3-methylglutaryl-coenzyme A synthase from Matricaria chamomilla

Abstract

3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) catalyzes the condensation of acetyl-CoA and acetoacetyl-CoA to form 3-hydroxy-3-methylglutaryl-CoA as the first committed enzyme in the mevalonate (MVA) pathway. HMGS plays an important role in the biosynthesis of the sesquiterpene, which is the main constituent of essential oil in Matricaria chamomilla. In this paper, a HMGS gene designated as McHMGS (GenBank Accession No. KU529970) was successfully cloned from M. chamomilla. The full-length cDNA of McHMGS was 1495-bp and contained a 1374-bp open reading frame. It encoded a 458-amino-acid protein with a calculated molecular weight of about 50.7 kDa and isoelectric point of 5.69. Sequence comparison revealed that McHMGS showed extensive homology with HMGSs from other plant species. Phylogenetic tree analysis indicated that McHMGS is clustered with the HMGS of Asteraceae in the dicotyledoneae clade. Further functional complementation of McHMGS in hmgs-deficient mutant yeast strain YSC6274 demonstrated that cloned McHMGS cDNA encodes a functional HMGS and mediates the MVA biosynthesis in yeasts. The tissue expression pattern analysis revealed that McHMGS expression level is highest in the flowers and lowest in the stems. Quantitative real-time PCR analysis showed that the expression of McHMGS was induced by MeJA, and the expression level is highest 24 h after induction. The characterization and expression of McHMGS can help in further studying the role of McHMGS gene in the biosynthesis of sesquiterpene in M. chamomilla.

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