Bacopa monniera recombinant mevalonate diphosphate decarboxylase: Biochemical characterization

Bacopa monniera recombinant mevalonate diphosphate decarboxylase: Biochemical characterization

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 Es...

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Bibliographic Details
Published in:International journal of biological macromolecules Vol. 79; pp. 661 - 668
Main Authors: Abbassi, Shakeel J., Vishwakarma, Rishi K., Patel, Parth, Kumari, Uma, Khan, Bashir M.
Format: Journal Article
Language:English
Published: Netherlands Elsevier B.V 01-08-2015
Subjects:
Bacopa - chemistry
Bacopa - enzymology
Carboxy-Lyases
Cations, Divalent
Cloning, Molecular
Enzyme Assays
Enzyme Stability
Escherichia coli - genetics
Escherichia coli - metabolism
Gene Expression
Hemiterpenes - chemistry
Hemiterpenes - metabolism
Hydrogen-Ion Concentration
Kinetics
Magnesium - chemistry
Mevalonate diphosphate decarboxylase
Mevalonic Acid - analogs & derivatives
Mevalonic Acid - chemistry
Mevalonic Acid - metabolism
Molecular Weight
Organophosphorus Compounds - chemistry
Organophosphorus Compounds - metabolism
pH activity profile
Phylogenetic analysis
Plant Proteins - chemistry
Plant Proteins - genetics
Plant Proteins - metabolism
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Stability
Temperature
Phylogenetic analysis
Stability
Mevalonate diphosphate decarboxylase
pH activity profile
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Summary: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+.
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ISSN:0141-8130
1879-0003
DOI:10.1016/j.ijbiomac.2015.05.041