Cloning and characterization of glucosyltransferase cDNA from Eucalyptus perriniana cultured cells

Abstract

The Eucalyptus perriniana cultured cells are widely used to biotransform a variety of compounds. The glucosyltransferase activity of a crude protein extract of E. perriniana cultured cells was maximized when cell growth was in the pre-logarithmic to logarithmic phase. We cloned a cDNA encoding glucosyltransferase (EPGT) from E. perriniana cultured cells by RT-PCR using a degenerated primer and RACE-PCR. The cDNA contained an open reading frame encoding 467 amino acids with a calculated molecular mass of 51.6 kDa. The consensus sequence of the plant glucosyltransferases was included in the deduced amino acid sequence. The amino acid sequence of EPGT showed a high identity to glucosyltransferases from tobacco and petunia. The recombinant EPGT was expressed in Escherichia coli and its substrate specificity was examined using UDP-[U-¹⁴C] glucose. Cinnamic acid was the best sugar acceptor in the compounds tested.