Cloning and functional analysis of hydroxylase gene TcCYP725A22 from Taxus wallichiana var.chinensis

Based on our previous transcriptome analysis, a new P450 hydroxylase gene was identified and named TcCYP725A22 according to homological analysis. The TcCYP725A22 cDNA was 1500 bp in length and encoded 499 amino acids. Bioinformatics predicted that the TcCYP725A22 protein contained a transmembrane region and catalytically active domains but no signal peptide. Functional analysis of TcCYP725A22 was carried out in Saccharomyces cerevisiae strain WAT11. Western blotting results indicated that the target protein was expressed successfully in WAT11. Furthermore, LC-MS analysis showed that TcCYP725A22 expressed in yeast was able to transform taxusin to its hydroxyl derivative, indicating that it had catalytic activity.