The purine and pyrimidine metabolism of Tetrahymena pyriformis

The metabolism of purines and pyrimidines by the ciliated protozoan Tetrahymena was investigated with the use of enzymatic assays and radioactive tracers. A survey of enzymes involved in purine metabolism revealed that the activities of inosine and guanosine phosphorylase (purine nucleoside: orthophosphate ribosyltransferase, E.C. 2.4.2.1) were high, but adenosine phosphorylase activity could not be demonstrated. The apparent K_m for guanosine in the system catalyzing its phosphorolysis was 4.1 ± 0.6 × 10^?3 M. Pyrophosphorylase activities for IMP and GMP (GMP: pyrophosphate phosphoribosyltransferase, E.C. 2.4.2.8), AMP (AMP: pyrophosphate phosphoribosyltransferase, E.C. 2.4.2.7), and 6-mercaptopurine ribonucleotide were also found in this organism; but a number of purine and pyrimidine analogs did not function as substrates for these enzymes. The metabolism of labeled guanine and hypoxanthine by intact cells was consistent with the presence of the phosphorylases and pyrophosphorylases of purine metabolism found by enzymatic studies. Assays for adenosine kinase (ATP: adenosine 5'-phosphotransferase, E.C. 2.7.1.20) inosine kinase, guanosine kinase, xanthine oxidase (xanthine: O₂ oxidoreductase, E.C. 1.2.3.2), and GMP reductase (reduced-NADP: GMP oxidoreductase [deaminating], E.C. 1.6.6.8) were all negative. In pyrimidine metabolism, cytidine-deoxycytidine deaminase (cytidine aminohydrolase, E.C. 3.5.4.5), thymidine phosphorylase (thymidine: orthophosphate ribosyltransferase, E.C. 2.4.2.4), and uridine-deoxyuridine phosphorylase (uridine: orthophosphate ribosyltransferase, E.C. 2.4.2.3) were active; but cytidine kinase, uridine kinase (ATP: uridine 5'-phosphotransferase, E.C. 2.7.1.48), and CMP pyrophosphorylase could not be demonstrated.     

The Distribution of Tetrahymena pyriformis1

SYNOPSIS. This paper is a brief account of both amicronucleate and sexually active strains of Tetrahymena pyriformis and their distribution with some comments on their possible evolution.     

Degradation of Pure Aflatoxins by Tetrahymena pyriformis

Tetrahymena pyriformis W with nutrients, ca. 22 × 10⁶ cells, decreased the concentration of aflatoxin B₁ 58% in 24 hr and 67% in 48 hr. An unknown, bright-blue fluorescent substance was produced, with intensity about one-half that of the unchanged B₁, with an R_f of 0.52 compared with 0.59 for B₁ and 0.55 for B₂ on a thin-layer chromatography plate, and with an ultraviolet spectrum showing maxima of 253, 261, and 328 mμ. In a separate assay, the cells with nutrients did not degrade pure G₁. Starved, washed cells, ca. 11 × 10⁶, decreased the concentration of B₁ 50% in 10 hr, 70% in 22 hr, and 75% in 30 hr, producing the same unknown component. Ethyl alcohol, 1.96% (v/v), decreased cell populations and size, but the cells remained actively motile in broth plus the alcohol for 96 hr. In 72 hr, neither toxin (ca. 2 ppm) in combination with ethyl alcohol had more inhibitory effect on cell numbers, with or without nutrients, than was produced by alcohol alone. Aflatoxin B₁ had no observed effect on the viability of the starved cells for 30 hr or on the nourished cells for 72 hr. There was no noticeable effect of G₁ on the starved cells in 30 hr or on the nourished cells in 48 hr. After 72 hr with G₁ plus nutrients, many of the cells were round with blisters, nonmotile, and apparently dead or dying.