Purification and characterization of cinnamyl alcohol-NADPH-dehydrogenase from the leaf tissues of a basin mangrove Lumnitzera racemosa Willd.
Cinnamyl alcohol-NADPH-dehydrogenase (CAD), the marker enzyme of lignin biosynthesis was purified from the leaf tissues of a basin mangrove Lumnitzera racemosa by ammonium sulphate precipitation, followed by anion-exchange, gel filtration and affinity chromatography. The molecular mass of the CAD enzyme was determined as 89 kDa, by size elution chromatography. SDS-PAGE of CAD revealed two closely associated bands of 45 kDa and 42 kDa as heterogenous subunits. The optimum pH of CAD was found to be 4.0. Km for the substrates cinnamaldehyde, coniferaldehyde and sinapaldehyde was determined. Cinnamaldehyde showed higher Km value than sinapaldehyde and coniferaldehyde. The correlation of activity of CAD with the amount of lignin was found less significant in L. racemosa, compared to plant species of other habitats viz., mesophytes, xerophytes and hydrophytes, suggesting that CAD possibly exhibits physiological suppression due to the saline habitat of the plant.
Plant Biochemistry and Molecular Biology Unit, Department of Botany, University College, Thiruvananthapuram 695 034, Kerala, India.
K
K
Murugan
N S
NS
Arunkumar
C
C
Mohankumar
D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel; D029764:Combretaceae / Q000201:enzymology*; D003636:DEAE-Cellulose / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008031:Lignin / Q000737:chemistry; D009252:NADPH Dehydrogenase / Q000737:chemistry*; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000302:isolation & purification; D020005:Propanols / Q000737:chemistry / Q000302:isolation & purification*; D013997:Time Factors
D010940:Plant Proteins; D020005:Propanols; D008031:Lignin; D003636:DEAE-Cellulose; D009252:NADPH Dehydrogenase; C020722:cinnamyl alcohol