Cloning and Functional Analysis of the Candida glycerinogenes TRP1 Gene

Abstract:

Candida glycerinogenes WL2002-5 is an osmotolerant yeast used for the commercial production of glycerol. TRP1 gene of Candida glycerinogenes (CgTRP1), encoding phosphoribosyl anthranilate isomerase(PRA1) was cloned by complementation of the trp1 mutation of Saccharomyces cerevisiae W303-1A. Sequence analysis revealed the presence of a 735 bp open reading frame(ORF) encoding 244 amino acids protein, which shares 32.9 %~49.2% amino acids sequence similarity to PRAI proteins from other species of Saccharomycetales. Functional analysis reveal, high copy number of CgTRP1 can complement the trp1 mu-tation of S. cerevisiae completely, but low copy number of CgTRP1 can only partially complement the trp1 mutation. Downstream from CgTRP1 was found a gene highly similar to IPP1 gene which encoding inor-ganic pyrophosphatase. The nucleotides have been deposited in the GenBank database under accession No. EU078909.

Key words:Candida glycerinogenes, Phosphoribosyl anthranilate isomerase, trp1 mutation