Physiological role of S-formylglutathione hydrolase in C1 metabolism of the methylotrophic yeast Candida boidinii

Abstract

The methylotrophic yeast Candida boidinii exhibits S-formylglutathione hydrolase activity (FGH, EC 3.1.2.12), which is involved in the glutathione-dependent formaldehyde oxidation pathway during growth on methanol as the sole carbon source. The structural gene, FGH1, was cloned from C. boidinii, and its predicted amino acid sequence showed more than 60 % similarity to those of FGHs from Paracoccus denitrificans and Saccharomyces cerevisiae, and human esterase D. FGH from C. boidinii contained a C-terminal tripeptide, SKL, which is a type I peroxisometargeting signal, and a bimodal distribution of FGH between peroxisomes and the cytosol was demonstrated. The FGH1 gene was disrupted in the C. boidinii genome by one-step gene disruption. The fgh1D strain was still able to grow on methanol as a carbon source under methanollimited chemostat conditions with low dilution rates (D<0?05 h "1 ), conditions under which a strain with disruption of the gene for formaldehyde dehydrogenase (another enzyme involved in the formaldehyde oxidation pathway) could not survive. These results suggested that FGH is not essential but necessary for optimal growth on methanol. This is believed to be the first report of detailed analyses of the FGH1 gene in a methylotrophic yeast strain.

A Novel Fluorescent Sensor Protein for Visualization of Redox States in the Cytoplasm and in Peroxisomes

“…This observation seems to reflect the fact that the high-energy bonds of fatty acid thioesters are stabilized in peroxisomes during ␤-oxidation. On the other hand, during methanol metabolism, the reduced glutathione within peroxisomes is consumed through two critical reactions for methanol metabolism, i.e., by forming hydroxymethyl glutathione nonenzymatically and by being further oxidized to S-formylglutathione to metabolize formaldehyde (39) or by Pmp20-glutathione peroxidase to detoxify intraperoxisomal ROS (9).…”

Unique Regulation of the Active site of the Serine Esterase S-Formylglutathione Hydrolase

“…Esterase D, originally identified as a carboxyesterase in man with high activity toward 4-methylumbelliferyl acetate (MUA), was subsequently shown to be S-formylglutathione hydrolase (SFGH, EC 3.1.1.56), an enzyme catalysing the hydrolysis of S-formylglutathione to formic acid and glutathione. 4 SFGH is part of a pathway of formaldehyde detoxification conserved in prokaryotes and eukaryotes, 5 with the enzyme functionally characterised in Paracoccus denitrificans, 6 Saccharomyces cerevisiae, 7 Candida boidinii, 8 Escherichia coli 9 and A. thaliana. 10,11 Searches for homologues in other genome and EST databases show remarkable conservation in the sequence of SFGHs.…”

Proteomic analysis of mycelial proteins from Magnaporthe oryzae under nitrogen starvation

“…S-Formyl glutathione hydrolase is regulated by carbon and nitrogen source and is involved in the glutathione-dependent formaldehyde oxidation pathway. Its products can be used as carbon sources for growth (Yurimoto et al, 2003). Our results showed that the expression of S-formyl glutathione hydrolase in the mycelia was down-regulated under low nitrogen stress, suggesting that M. oryzae enhanced nitrogen metabolism in the mycelia and improved the synthetic ability of the desired carbon and nitrogen in order to ameliorate the adaptability of strains to adverse environmental conditions of nitrogen deficiency.…”