Isolation and sequence analysis of the gene encoding triose phosphate isomerase from Zygosaccharomyces bailii

Abstract

The ZbTPI1 gene encoding triose phosphate isomerase (TIM) was cloned from a Zygosaccharomyces bailii genomic library by complementation of the Saccharomyces cerevisiae tpi1 mutant strain. The nucleotide sequence of a 1.5 kb fragment showed an open reading frame (ORF) of 746 bp, encoding a protein of 248 amino acid residues. The deduced amino acid sequence shares a high degree of homology with TIMs from other yeast species, including some highly conserved regions. The analysis of the promoter sequence of the ZbTPI1 revealed the presence of putative motifs known to have regulatory functions in S. cerevisiae. The GenBank Accession No. of ZbTPI1 is AF325852.

The yeast : a new host for heterologous protein production, secretion and for metabolic engineering applications

“…In plasmid pZ 3 bT (Fig. The sequence was ampli¢ed by polymerase chain reaction (PCR) from the genomic DNA of the Z. bailii strain ISA 1307, and the primers were designed according to the literature [26]. The sequence was ampli¢ed by polymerase chain reaction (PCR) from the genomic DNA of the Z. bailii strain ISA 1307, and the primers were designed according to the literature [26].…”

“…1b), the S. cerevisiae TPI promoter was replaced with the endogenous TPI promoter from Z. bailii. The sequence was ampli¢ed by polymerase chain reaction (PCR) from the genomic DNA of the Z. bailii strain ISA 1307, and the primers were designed according to the literature [26]. Extraction of genomic DNA was performed according to the protocol published [36] and ampli¢cation was obtained annealing the oligonucleotides 5P-ATC GTA TTG CTT CCA TTC TTC TTT TGT TA-3P and 5P-TTT GTT ATT TGT TAT ACC GAT GTA GTC TC-3P.…”

Aerobic sugar metabolism in the spoilage yeast

“…Zygosaccharomyces bailii is able to catabolise weak acids in the complex mixture with sugars present in some foods [2,3] and its 'spoilage' ability has been the subject of some studies in recent years [4^8]. The sole gene implicated in sugar metabolism isolated so far is the gene encoding triose phosphate isomerase [12]. Z. bailii is known as a fructophilic yeast: fructose is transported by a speci¢c high-capacity system, while glucose is transported by a lower-capacity system, partially inactivated by fructose [9].…”

“…Aerobic alcoholic fermentation in Z. bailii has been observed [10] and it was recently reported that Z. bailii is also able to grow anaerobically in complex media [11]. The sole gene implicated in sugar metabolism isolated so far is the gene encoding triose phosphate isomerase [12].…”

Molecular cloning and sequence analysis of the Zygosaccharomyces bailii HIS3 gene encoding the imidazole glycerolphosphate dehydratase

“…Up to now, the only example in the literature that shows an application of this genetic knowledge concerns the expression of alkaline protease from Aspergillus oryzae in Z. rouxii (Ogawa et al , 1990). With regard to Z. bailii , only recently has some information about its genetic manipulation, the construction of a genomic library and the cloning of some genes, become available (Mollapour and Piper, 2001; Merico et al , 2001; Rodrigues et al , 2001). In our laboratory, we recently established the minimal tools for developing Z. bailii as a new host for the production of heterologous proteins (Brambilla et al , 2000).…”