Purification and characterization of a catechol 1,2-dioxygenase from a phenol degrading Candida albicans TL3

Abstract

A eukaryotic catechol 1,2-dioxygenase (1,2-CTD) was produced from a Candida albicans TL3 that possesses high tolerance for phenol and strong phenol degrading activity. The 1,2-CTD was purified via ammonium sulfate precipitation, Sephadex G-75 gel filtration, and HiTrap Q Sepharose column chromatography. The enzyme was purified to homogeneity and found to be a homodimer with a subunit molecular weight of 32,000. Each subunit contained one iron. The optimal temperature and pH were 25 degrees C and 8.0, respectively. Substrate analysis showed that the purified enzyme was a type I catechol 1,2-dioxygenase. This is the first time that a 1,2-CTD from a eukaryote (Candida albicans) has been characterized. Peptide sequencing on fragments of 1,2-CTD by Edman degradation and MALDI-TOF/TOF mass analyses provided information of amino acid sequences for BLAST analysis, the outcome of the BLAST revealed that this eukaryotic 1,2-CTD has high identity with a hypothetical protein, CaO19_12036, from Candida albicans SC5314. We conclude that the hypothetical protein is 1,2-CTD.

The old 3-oxoadipate pathway revisited: New insights in the catabolism of aromatics in the saprophytic fungus Aspergillus nidulans

“…These genes, namely AN4532, AN8998, AN0764 and AN9363, may be classed in three sub-groups of a single evolutionary lineage family, namely catechol 1,2-dioxygenase, hydroxyquinol 1,2-dioxygenase and protocatechuate 3,4-dioxygenase. A member of the first sub-group, previously characterised and identified in Candida albicans (Tsai and Li, 2007), is orthologous to AN4532 and AN8998 (paralogous genes). Candida parapsilosis utilises benzoate (and derivatives) through a hydroxyquinol variant of the 3-oxoadipate pathway, hence its single catechol dioxygenase-like gene belongs to the second sub-group (Holesova et al, 2011), which is orthologous to AN0764 and AN9363.…”

“…Catechol conversion to cis,cis-muconate in C. albicans involves a catechol 1,2-dioxygenase (Tsai and Li, 2007), of which the encoding gene shows the highest sequence identity with AN4532. As expected, the last underwent major induction in the salicylate medium, at both protein (11 spots, see Supplemental Fig.…”

Catechol 1,2-dioxygenase from α-naphthol degrading thermophilic Geobacillus sp. strain: purification and properties

“…Enzymes of the various aromatic hydrocarbons biodegradation pathway, including C1,2O, are inducible in microorganisms [2][3][4]. C1,2O have been purified from a variety of organisms comprising Pseudomonas, Alcaligenes, Ralstonia, Rhodococcus, Acinetobacter and Candida albicans [5][6][7][8][9][10].…”

Effect of Piperonyl Butoxide and Silver Nitrate on Degradation Pathways of n-octadecane by Fusarium sp. F092

“…The effect of inhibition of monooxygenase and dioxygenase was determined as reported previously (Tsai & Li, 2007;Mori et al, 2009). The cultures were incubated for 15 days, and the degradation was examined.…”

“…F092 could utilize aliphatic fraction in crude oil as carbon and energy sources needs to be further investigated. Degradation process is usually closely relative to the enzyme production, which could be also examined by addition of an inducer or inhibitor for a specific enzyme to the culture (Mori & Kondo, 2002;Tsai & Li, 2007;Mori, Nakamura, & Kondo, 2009). The objectives of this study were: 1) to investigate the capability of Fusarium sp.…”