Cloning of a cDNA encoding a NAD-dependent formate dehydrogenase involved in oxalic acid metabolism from the white-rot fungus Ceriporiopsis subvermispora and its gene expression analysis.
The authors have proposed previously that intracellular degradation of oxalic acid via formate to CO(2) occurs in the white-rot fungus Ceriporiopsis subvermispora. The formate degradation is catalyzed by NAD-dependent formate dehydrogenase (CsFDH). In this study, two cDNAs named CsFDH1 and CsFDH2 encoding CsFDH were cloned. Each cDNA consisting of 1077 bp encodes a mature protein composed of 358 amino acid residues. The amino acid sequences of the deduced CsFDH1 and CsFDH2 showed 99% identity to each other. The predicted molecular mass for each was 39.3 kDa, which was similar to that of CsFDH purified from the vegetative mycelia of Ceriporiopsis subvermispora (purified-CsFDH). The recombinant CsFDH1 and CsFDH2 expressed by Escherichia coli showed FDH activity with similar characteristics to the purified CsFDH. However, the amount of CsFDH1 transcript from the vegetative mycelia was 236-691 times greater than that of CsFDH2. Therefore, the results strongly suggest that CsFDH1, as compared with CsFDH2, predominantly contributes to the production of the purified CsFDH.
Watanabe
Tomoki
T
Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto, Japan.
Fujiwara
Tadashi
T
Umezawa
Toshiaki
T
Shimada
Mikio
M
Hattori
Takefumi
T
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005560:Formate Dehydrogenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019815:Oxalic Acid / Q000378:metabolism*; D020072:Polyporales / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D018076:DNA, Complementary; D005656:Fungal Proteins; D011994:Recombinant Proteins; D019815:Oxalic Acid; D005560:Formate Dehydrogenases