Factors affecting division rhythmicity in Euglena gracilis

In phototrophic culture of Euglena gracilis, good synchrony was found only under rather restricted programs of light-dark cycles, and rather narrow ranges of temperature and light intensity, when cultures were flushed with air fortified with adequate amounts of CO₂. When flushed with air alone, CO₂ was found to be limiting, and while cell divisions were rhythmic, less than a doubling of cell number occurred in division bursts. With air as gas phase, rhythmic division activity was maintained over wide ranges of temperature, light intensity, and the ratio of light:dark in a given program; all these factors affected the amplitude of the division burst, however.     

Factors affecting pH-dependent photo-inhibition of division in Euglena gracilis

Visible light of moderate intensity (1200 ft-cd) can severely inhibit cell division of a non-photosynthetic mutant of Euglena gracilis when growth is supported by butanol, ethanol, or fumarate as sole carbon source. The degree of inhibition is pH dependent, being greatest at pH 4 to 5. A wide variety of other carbon sources permitted growth in the light without inhibition.     

Phenolic acids inhibit chloroplast mutagenesis in Euglena gracilis

The mutagenicity (bleaching activity) of ofloxacin (43 microM) and acridine orange (AO) (13.5 microM) in Euglena gracilis is inhibited by plant phenolics. Caffeic acid (CA), p-coumaric acid (PCA), ferulic acid (FA) and gentisic acid (GA) (25, 50, 100 and 250 microM) exhibited a significant concentration-dependent inhibitory effect against ofloxacin-induced mutagenicity, which was very effectively eliminated by the highest concentration of all four of those phenolic acids. The mutagenicity of AO was also significantly reduced in the presence of CA, PCA and FA (25, 50, 100 and 250 microM). However, GA exhibited no significant activity, even at the concentration of 250 microM. Based on the UV spectrophotometric measurements, we suggest that the antimutagenic effect of CA, PCA, FA and GA resulted from the scavenging of reactive oxygen species (ROS) produced by ofloxacin. On the other hand, the reduction of AO-induced mutagenicity correlates with the binding capabilities of CA, PCA and FA, with the exception of GA.     

Waxmonoester fermentation in Euglena gracilis T. Factors favouring the synthesis of odd-numbered fatty acids and alcohols

Waxmonoester fermentation at the expense of endogenous paramylon was followed in the dark in autotrophically grown Euglena gracilis. With reduced oxygen tension and decreasing O₂-consumption rates the proportion of odd-numbered fatty acids and alcohols increased up to a molar ratio of nearly 1:1 under strictly anaerobic conditions. Labelled ¹⁴CO₂, succinate and propionate were incorporated into odd-numbered fatty acids and alcohols 11 to 33 times faster than in even-numbered chains. The electron-flow inhibitor rotenone diminished waxester formation in total, but especially CO₂ fixation and the synthesis of odd-numbered chains, without impeding anaerobic carbohydrate breakdown. These findings are indicative for propionyl-CoA as an intermediate in the synthesis of odd-numbered chains. Its probable synthesis in the methylmalonyl-CoA pathway is discussed with regard to energetics.Abbreviation CCCP carbonylcyanide m-chlorophenyl hydrazone     

Heteroxanthin in Euglena gracilis

Summary 1. From a large scale preparation of Euglena gracilis, strain Z, besides the acetylenic pigments diatoxanthin and diadinoxanthin and the allene neoxanthin, an additional acetylenic xanthophyll has been isolated. 2. Mass and IR spectra and chemical reactions showed typical patterns of heteroxanthin from Vaucheria. 3. The pigment was transformed into diadinochrome-isomers with acidified acetone. 4. A partial synthesis of heteroxanthin from diadinoxanthin by LiAlH₄-reduction is described, confirming the structure proposed by Strain. 5. The identity of heteroxanthin with the trollein—like pigment described for Euglena is discussed.     

Proteolysis in Euglena gracilis

Endoproteolytic activities (EC 3.4.22. and 23.) of cell-free extracts of Euglena gracilis, measured by autolysis and azocaseinolysis, vary considerably during the culture growth cycle. They are high in the lag phase, drop sharply up to the mid-logarithmic phase, and then rise again reaching the initial high levels in the stationary phase. This pattern has been observed for both the soluble and the particulate proteolytic activities of four cell types differing with regard to the developmental state of the chloroplast: dark-grown, light-induced, and light-grown wild-type cells, as well as light-grown apoplastic W₃BUL mutant cells, all on a glucose-based medium. Therefore, the activity of the main intracellular proteinases is neither directly nor indirectly light-regulated, but seems to be controlled by the availability of nutrients. Endogenous inhibitors of proteinases could not be detected. Cysteine proteinase activity has been found in the soluble and the particulate fractions, but aspartic proteinase activity in the latter ones only. Different cysteine proteinases may be present in the two fractions, during the different growth phases, and in the four cell types studied.Abbreviations CBB Coomassie Brilliant Blue G-250 - DFP diisopropyl fluorophosphate - EDTA disodium ethylendiaminetetraacetic acid - E-64 l-transepoxysuccinyl-leucyl-amido(4-guanidino)butane - Iog phase logarithmic growth phase - MET 2-mercaptoethanol - PMSF phenylmethylsulfonyl fluoride - Z benzyloxycarbonyl Paper I of this series is Krauspe and Scheer (1986). A preliminary publication appeared (Krauspe et al. 1982)     

Antimutagenicity in Euglena gracilis

The genotoxic effect of N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and furadantine (Fu) was significantly decreased by standard antimutagens (ascorbic acid, α-tocopherol, chlorophyllin and sodium selenite) in the unicellular flagellate Euglena gracilis. The effects of these compounds were verified also by a bacterial test in which three strains of Salmonella typhimurium, TA97, TA100 and TA102, were used. The above compounds were antimutagenic in strains of bacteria used, except for chlorophyllin which had no effect on strain TA102.     

Incorporation of labelled amino acids by nuclei isolated from Euglena gracilis

Nuclei isolated from the lower eukaryote, Euglena gracilis, incorporate labelled amino acids into hot-TCA-insoluble material under conditions of incubation similar to those used for higher eukaryotes. Optimal temperature for amino acid incorporation by Euglena nucleic is 25° to 30°C. Optimal pH is 5.0. ATP stimulates incorporation only to a small extent. Evidence is presented that the product synthesized is protein.     

Peroxidative Activity in Euglena gracilis

Cell-free homogenates of Euglena gracilis contain very low levels of catalase activity as compared to higher plants and some other algae. Purified Euglena cytochrome c acts catalytically as a peroxidase. The observed catalytic activity of cytochrome c in extracts from heterotrophically grown cells was more than enough to account for the observed rates of hydrogen peroxide destruction. The peroxidative activity of Euglena cytochrome c was completely inhibited by 20 mm 3-amino-1,2,4-triazole.     

Thiamin uptake in Euglena gracilis

Thiamin uptake has been investigated in Euglena gracilis Z. This protozoon possessed an active transport system for thiamin with a Km value of 17 nM and a Vmax value of 7.8 pmol per 10(6) cells per min. Thiamin uptake was dependent on pH and temperature, but not on exogenous glucose as an energy source. Oxythiamin and pyrithiamin were competitive inhibitors with Ki values of 33 nM and 15 nM, respectively. Thiamin monophosphate, thiamin pyrophosphate, thiamin triphosphate, heteropyrithiamin, quinolinothiamin, thiamin chloride and amprolium inhibited uptake. Inhibition of thiamin uptake by various metabolic inhibitors and anaerobiosis suggest that thiamin uptake requires an energy source generated by respiration and glycolysis.     

Biosynthesis of polyamines in Euglena gracilis

In Euglena gracilis Z the biosynthesis of spermidine and spermine closely resembles the pathways occurring in mammalian tissues and in most microorganisms. l-Ornithine and not l-arginine, as is the case in most plants, is the main precursor of putrescine, and S-adenosylmethionine donates the propylamino moiety for the biosynthesis of spermidine and spermine. Cell-free extracts of Euglena synthesized sym-norspermidine and sym-norspermine from 1,3-diaminopropane and labelled S-adenosylmenthionine. The synthases for the biosynthesis of these two polyamines have a pH optimum of 7.6, like that of spermidine and spermine synthases. Ion exchange chromatography showed two peaks corresponding to the retention times of 2,4-diaminobutyric acid and 1,3-diaminopropane, lower homologues of ornithine and putrescine, respectively. Experiments with dl-2,4-diaminobutyric acid-[4-¹⁴C] did not result in significant incorporation of the label into 1,3-diaminopropane.     

Erythromycin-Induced Bleaching of Euglena gracilis

SYNOPSIS. Erythromycin bleaches Euglena gracilis in a manner resembling that of streptomycin. Erythromycin-bleached substrains have been cultivated 16 months in light on erythro-mycin-free media without greening. Bleached substrains were obtained only if erythromycin was added to actively growing cultures: erythromycin did not bleach if added during the stationary phase of growth of green cultures.