In silico Characterisation of Conserved Functional Domain and Isolation of Coniferyl Aldehyde Dehydrogenase Gene from Pseudomonas nitroreducens Jin1

Coniferyl aldehyde dehydrogenase gene (calB) oxidises coniferyl aldehyde into ferulic acid, the last step in the metabolic pathway of eugenol to ferulic acid. It occurs in various bacterial strains like Pseudomonas nitroreducens Jin1, Ralstonia eutropha, Escherichia coli, Pseudomonassp. HR199, Rhodococcus opacus. Coniferyl aldehyde produced as the intermediate product by the biotransformation of eugenol, is oxidised to ferulic acid, NADPH and H+. Aldehyde dehydrogenase-related bacterial sequences were aligned and using insilico methodology analysed to determine functionally important conserved residues in this protein. A total of 16 residues have been identified to be conserved in this protein. calB gene from P.nitroreducens Jin1 located in region 107290-108732 (1442bp) was found to contain all the functionally important residues and hence was selected for isolation.