Cloning of the transketolase gene from erythritol-producing yeast Candida magnoliae

Abstract

The entire nucleotide sequence of the TKL1 gene encoding transketolase (TKL) in an erythritolproducing yeast of Candida magnoliae was determined by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed an open reading frame of C. magnoliae TKL1 (CmTKL1) that spans 2,088 bp and encodes 696 amino acids, sharing 61.7% amino acid identity to Kluyveromyces lactis TKL. Functional analysis showed that CmTKL1 complemented a Saccharomyces cerevisiae tkl1 tkl2 double mutant for growth in the absence of aromatic amino acids and restored transketolase activity in this mutant. An enzyme activity assay and RT-PCR revealed that the expression of CmTKL1 is induced by fructose, H2O2, and KCl. The GenBank accession number for C. magnoliae TKL1 is KF751756.