Purification and Properties of a Catechol Methyltransferase of the Yeast Candida tropicalis

Abstract

In an effort to investigate catechol methyltransferase activity in sources other than mammalian tissues and cells, a high level of enzyme activity was found in the yeast fungus Candida tropicalis CBS 94. Partial purification of the enzyme (approx. 550 fold with a recovery of 7%) could be achieved by using ion-exchange and gel filtration techniques. The molecular weight was estimated at 32,000 ± 2,000 by gel filtration on Sephadex G-100. In isoelectric focusing experiments on Sephadex G-75 the enzyme exhibited a pl-value o f 5.0 ± 0.1. In contrast to catechol methyltransferase from various mammalian tissues the enzyme activity was prepared from the pH 5-sediment. The substrate specifity is comparable to other catechol methyltransferases.

Kinetics and inhibition studies of catechol O-methyltransferase from the yeast Candida tropicalis

“….m JLM Ultrogel AcA44, and by isoelectric focusing on Sephadex G-75 as described earlier (27). The kinetic data for yeast COMT presented here are also compatible with the assumption that there is only one enzyme activity, because the linearity of double-reciprocal plots over a wide range of esculetin concentrations showed the absence of contamination by other molecular forms in the COMT preparation.…”

“…Yeast COMT was prepared as described earlier (27) COMT, and 4 ,uM esculetin to produce a final volume of 0.5 ml. After incubation for 30 min at 37°C, the reaction was terminated by heat denaturation.…”

“…By immunocytochemical techniques mammalian COMT was localized extraneuronally in the brain (19) and in the liver parenchyma at and around the glycogen granules (28). COMT activity was also found in several strains of the facultative pathogenic yeast Candida tropicalis and was highly purified (27,30). The enzyme was characterized immunocytochemically by light and electron microscopy and localized in the outer layer of the cell wall and at the plasma membrane (29).…”

Catechol‐O‐methyltransferase and Its Inhibitors in Parkinson's Disease

“…Queries at the National Center for Biotechnology Information databases (Wheeler et al 2006) show that the COMT gene has been found and characterized in a variety of species, such as Homo sapiens , Pan troglodytes , Canis familiaris , Rattus norvegicus , Mus musculus , Equus caballus, Gallus gallus , and Schizosaccharomyces pombe , and is conserved in the fungi/metazoa group. COMT is present in prokaryotes and eukaryotes, namely bacteria (Dhar et al 2000; Kim et al 2004Vilbois et al 1994), yeast (Veser 1987; Veser et al 1979), plants (Guldberg and Marsden 1975; Legrand et al 1976) and animals (Guldberg and Marsden 1975), and invertebrates and vertebrates. In mammals COMT is present in two molecular forms: a soluble form (S‐COMT) and in another form associated with membranes (MB‐COMT) (Borchardt et al 1974; Grossman et al 1985; Jeffery and Roth 1984).…”

Functional and structural characterization of a novel catechol‐O‐methyltransferase from Schizosaccharomyces pombe

“…In addition to having vital roles in physiological processes, COMTs also have essential application values in industry. Many of their reaction products possess medicinal values, for example, ferulic acid is a catalysate of caffeic acid (catalyzed by a COMT homolog from Medicago sativa L ) and serves as a necessary material for cardiovascular drug production, and scopoletin is a COMT catalysate of esculetin and functions as a drug ingredient due to its antiviral and anti‐inflammatory properties. A COMT homolog from alfalfa can also catalyze caffeyl‐alcohol to form coniferyl‐alcohol , which is one of three basic components of lignin that has attracted great attention due to its high polymer material properties.…”