Primary structure analysis and functional expression of erythrose reductases from erythritol-producing fungi (Trichosporonoides megachiliensis SNG-42).
NADPH-dependent erythrose reductases (ERs) in erythritol-producing fungi, Trichosporonoides megachiliensis SNG-42, catalyze the reduction of D-erythrose. We previously characterized the biochemical properties of three isozymes of ERs (ER-I, ER-II, and ER-III). Using internal amino acid sequences of ER-III and ER-I with peptide mapping, we cloned three cDNAs (er1, 1121-bp (AB191474); er2, 1077-bp (AB191475); er3, 1119-bp (AB191476)). The er3 cDNA encoded a polypeptide 36,044 Da, and its deduced amino acid sequence was same as that of the native ER-III. The three recombinant enzymes expressed in Escherichia coli were purified to homogeneity. The recombinant enzymes of ER1, ER2, and ER3 showed similar electrophoretic properties to that of the native ER-I, ER-II, and ER-III on SDS- and Native- but not on IEF-PAGE. All three recombinant enzymes showed substrate specificity towards C-4 and C-3 aldehydes similar to that of the native ER-III. These results strongly suggest that cloned er1, er2, and er3 cDNAs encode erythrose reductases.
Ookura
Tetsuya
T
National Food Research Institute, Ibaraki. ookura@nfri.affrc.go.jp
Azuma
Keiko
K
Isshiki
Kenji
K
Taniguchi
Hajime
H
Kasumi
Takafumi
T
Kawamura
Yukio
Y
D000449:Aldehyde Reductase / Q000737:chemistry* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001203:Ascomycota / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004896:Erythritol / Q000096:biosynthesis*; D015870:Gene Expression; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000449:Aldehyde Reductase; D004896:Erythritol