Bacopa monniera recombinant mevalonate diphosphate decarboxylase: Biochemical characterization

Shakeel J. Abbassi, Rishi K. Vishwakarma, Parth Patel, Uma Kumari, Bashir M. Khan

Research output: Contribution to journalArticlepeer-review

Abstract

Mevalonate diphosphate decarboxylase (MDD; EC 4.1.1.33) is an important enzyme in the mevalonic acid pathway catalyzing the Mg2+-ATP dependant decarboxylation of mevalonate 5-diphosphate (MVAPP) to isopentenyl diphosphate (IPP). Bacopa monniera recombinant MDD (BmMDD) protein was overexpressed in Escherichia coli BL21 (DE3) strain and purified to apparent homogeneity. Km and Vmax for MVAPP were 144μM and 52Umg-1 respectively. The values of turnover (kcat) and kcat/Km for mevalonate 5-diphosphate were determined to be 40s-1 and 2.77×105M-1s-1 and kcat and kcat/Km values for ATP were found to be 30s-1 and 2.20×104M-1s-1, respectively. pH activity profile indicated the involvement of carboxylate ion, lysine and arginine for the activity of enzyme. The apparent activation energy for the BmMDD catalyzed reaction was 12.7kJmol-1. Optimum pH and temperature for the forward reaction was found to be 8.0 and 45°C. The enzyme was most stable at pH 7 at 20°C with the deactivation rate constant (Kd*) of 1.69×10-4 and half life (t1/2) of 68h. The cation studies suggested that BmMDD is a cation dependant enzyme and optimum activity was achieved in the presence of Mg2+.

Original languageEnglish (US)
Pages (from-to)661-668
Number of pages8
JournalInternational Journal of Biological Macromolecules
Volume79
DOIs
StatePublished - Aug 1 2015

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Economics and Econometrics
  • Energy(all)

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