Substrate inhibition inPseudomonas oxalaticus OX1: a kinetic study of growth inhibition by oxalate and formate using extended cultures

Pseudomonas oxalaticus OX1 has been grown in a mineral salts medium with oxalate or formate as the sole source of carbon and energy. At concentrations of these substrates above 50mm inhibition of growth was indicated by a long and variable lag phase in batch culture. This inhibition was further studied by estimating maximum specific growth rates at different substrate concentrations using the extended culture technique for control of the substrate concentration. With formate, inhibition became apparent at substrate concentrations above 20mm, whereas oxalate inhibited growth at concentrations above 15mm. Complete inhibition was not observed even at concentrations of 100mm. A number of inhibition functions were fitted with the experimental data using computer analysis. The results indicated that the Haldane equation was the simplest function to describe quantitatively the kinetics of the observed substrate inhibition. Studies on the rate of oxygen uptake at different concentrations of oxalate indicated that respiration was much more sensitive to inhibition than growth. However with formate, inhibition of respiration was not observed up to concentrations of 50mm, indicating that different mechanisms may underlie the observed growth inhibition by the two substrates.     

The effect of some environmental factors on the production of L-DOPA by alginate-entrapped cells of Mucuna pruriens

In-vitro-grown cells of Mucuna pruriens, immobilized in calcium-alginate gels, were able to transform the precursor L-tyrosine into L-dihydroxyphenylalanine (L-DOPA). After the immobilization in alginate the plant cells released 90% of the produced L-DOPA into the medium; supplementation of the medium with calcium inhibited both the transformation of L-tyrosine into L-DOPA and the release of L-DOPA into the medium. Continuous illumination of the beads had a slight beneficial effect on the synthesis of L-DOPA. A simple production medium for the transformation of L-tyrosine into L-DOPA was designed. This medium contained only sucrose and sodium chloride as osmotic stabilizers, a low concentration of calcium chloride for stabilization of the alginate beads, and L-tyrosine as the precursor.     

l-Carnitine is essential to β-oxidation of quarried fatty acid from mitochondrial membrane by PLA2

Mitochondrial β-oxidation is an important system involved in the energy production of various cells. In this system, the function of l-carnitine is essential for the uptake of fatty acids to mitochondria. However, it is unclear whether or not endogenous respiration, ADP-induced O₂ consumption without substrates, is caused by l-carnitine treatment. In this study, we investigated whether l-carnitine is essential to the β-oxidation of quarried fatty acids from the mitochondrial membrane by phospholipase A₂ (PLA₂) using isolated mitochondria from the liver of rats. Intact mitochondria were incubated in a medium containing Pi, CoA and l-carnitine. The effect of l-carnitine treatment on ADP-induced mitochondrial respiration was observed without exogenous respiratory substrate. Increase in mitochondrial respiration was induced by treatment with l-carnitine in a concentration-dependent manner. Treatment with rotenone, a complex I blocker, completely inhibited ADP-induced oxygen consumption even in the presence of l-carnitine. Moreover, the l-carnitine dependent ADP-induced mitochondrial oxygen consumption did not increase when PLA₂ inhibitors were treated before ADP treatment. The l-carnitine-dependent ADP-induced oxygen consumption did contribute to ATP productions but not heat generation via an uncoupling system. These results suggest that l-carnitine might be essential to the β-oxidation of quarried fatty acids from the mitochondrial membrane by PLA₂.     

The systematic significance of seed morphology and anatomy in thePortulacaceae (Centrospermae)

Nyananyo B. L. (1988): The systematic significance of seed morphology and anatomy in thePortulacaceae (Centrospermae).—Folia Geobot. Phytotax., Praha, 23: 275–279.—The seeds of 115 species in eighteen genera of the familyPortulacaceae were examined with the scanning electron microscope. Longitudinal sections of seeds of representatives of all the genera were examined with the light microscope. Two broad groups based on shape of the seeds, ornamentation of the external surface of their testa, and the position of the embryo relative to the endosperm were recognised. The first group comprising three genera:Anacampseros L.,Grahamia Gillies andTalinopsis A. Gray, have seeds which are pyriform with wings and trichomes, and an embryo that is more or less parallel to the endosperm, while the second of fifteen genera:Calandrinia H.B.K.,Calyptridium Nuttall,Calyptrotheca Gilg,Ceraria Pearson etStephens,Claytonia L.,Lenzia Philippi,Lewisia Pursh,Lyallia Hooker fil.,Montia L.,Portulaca L.,Portulacaria Jacquin,Silvaea Philippi,Talinella Baillon,Talinum Adanson, andWangerinia Franz, are reniform without wings and with an embryo that is horse-shoe shaped around the endosperm.     

The effect of N-nitroso-N-methylurea,buthylmethane sulphonate and X-rays on the germination and production of chlorophyll mutations in einkorn wheat

Grains ofTriticum monococcum L. var.sofianum Körn. were treated with 0·1mm, 0·2mm and 0·3mm solutions of N-nitroso-N-methylurea (MNH), with 0·03m solution of buthylmethane sulphonate (BMS) and with X-rays in doses of 5 000r and 10 000r. The germination and development of individual colors of chlorophyl mutants were observed by the system developed byLamprecht (1960). All the mutants induced were classified according to their color changes into three main categories-homogenous unicolor, homogenous multicolor and heterogenous multicolor. In the last type the colors of individual leaves of the same plant varied. Anthocyanin mutations “albina” and “albino-transvirgata” sometimes coincided with the chlorophyll mutations. Some chlorophyll mutations showing complicated groups of colors appeared which were beyond the scale of classification by ordinary systems. The largest proportion in the spectrum of chlorophyll mutations, induced by MNH and X-rays was occupied by mutations of thealbina type. The broadest mutation spectrum in our experiments was induced by the application of 0·3mm MNH. The doses of X-rays used induced relatively higher numbers ofalbina-type chlorophyll mutations than MNH and BMS. In our experiments we succeeded in inducing on medium size samples ofTriticum monococcum L. var.sofianum Körn not only almost all types of chlorophyll mutations, induced byFujii (1960, 1962) andMatsumura (1960), but in addition also a great number of other even more complicated chlorophyll mutations, which have never been previously described inTriticum monococcum. L.     

l-Proline uptake in Saccharomyces cerevisiae mitochondria can contribute to bioenergetics during nutrient stress as alternative mitochondrial fuel

l-Proline (pyrrolidine-2-carboxylic acid) is a distinctive metabolite both biochemically and biotechnologically and is currently recognized to have a cardinal role in gene expression and cellular signaling pathways in stress response. Proline-fueled mitochondrial metabolism involves the oxidative conversion of l-Proline to l-Glutamate in two enzymatic steps by means of Put1p and Put2p that help Saccharomyces cerevisiae to respond to changes in the nutritional environment by initiating the breakdown of l-Proline as a source for nitrogen, carbon, and energy. Compartmentalization of l-Proline catabolic pathway implies that extensive l-Proline transport must take place between the cytosol where its biogenesis via Pro1p, Pro2p, Pro3p occurs and mitochondria. l-Proline uptake in S. cerevisiae purified and active mitochondria was investigated by swelling experiments, oxygen uptake and fluorimetric measurement of a membrane potential generation (ΔΨ). Our results strongly suggest that l-Proline uptake occurs via a carried-mediated process as demonstrated by saturation kinetics and experiments with N-ethylmaleimide, a pharmacological compound that is a cysteine-modifying reagent in hydrophobic protein domains and that inhibited mitochondrial transport. Plasticity of S. cerevisiae cell biochemistry according to background fluctuations is an important factor of adaptation to stress. Thus l-Proline → Glutamate route feeds Krebs cycle providing energy and anaplerotic carbon for yeast survival.     

Fermentation of tartrate enantiomers by anaerobic bacteria,and description of two new species of strict anaerobes,Ruminococcus pasteurii and Ilyobacter tartaricus

Enumerations of tartrate-fermenting anaerobic bacteria with l-, d-, and m-tartrate as substrates revealed that l-tartrate fermenters outnumbered d- and m-tartrate fermenters by one to three orders of magnitude in all three anoxic environments studied. Highest numbers of tartrate-fermenting bacteria were found in freshwater creek sediments, less in polluted marine channels, and lowest numbers in anoxic sewage digestor sludge. Prevailing bacteria were isolated on every tartrate enantiomer. They all degraded tartrates via oxaloacetate. d- and m-tartrate-fermenting anaerobes were able to ferment l-tartrate as well, and were assigned to the genera Bacteroides, Acetivibrio, and Ilyobacter. l-Tartrate-fermenting anaerobes only utilized this enantiomer, and were characterized in more detail. Fermentation products on tartrate, citrate, pyruvate, and oxaloacetate were acetate, formate, and carbon dioxide. On fructose and glucose, also ethanol was formed. Freshwater isolates were Gram-positive cocci with large slime capsules, and were described as a new species, Ruminococcus pasteurii. Saltwater isolates were Gram-negative short rods, and were also described as a new species, Ilyobacter tartaricus. The guanosine-plus-cytosine content of the DNA was 45.2% and 33.1%, respectively.     

Properties, metabolisms, and applications of l-proline analogues

Due to the unique role of l-proline in the folding and structure of protein, a variety of synthetic proline analogues have been developed. l-Proline analogues have been proven to be valuable reagents for studying cellular metabolism and the regulation of macromolecule synthesis in both prokaryotic and eukaryotic cells. In addition to these fundamental researches, they are useful compounds for industrial use. For instance, microorganisms that overproduce l-proline have been obtained by isolating mutants resistant to l-proline analogues. They are also promising candidates for tuning the biological, pharmaceutical, or physicochemical properties of naturally occurring or de novo designed peptides. Among l-proline analogues, l-azetidine-2-carboxylic acid (l-AZC) is a toxic non-proteinogenic amino acid originally found in lily of the valley plants and trans-4-hydroxy-l-proline (4-l-THOP) is the most abundant component of mammalian collagen. Many hydroxyprolines (HOPs), such as 4-l-THOP and cis-4-hydroxy-l-proline (4-l-CHOP), are useful chiral building blocks for the organic synthesis of pharmaceuticals. In addition, l-AZC and 4-l-CHOP, which are potent inhibitors of cell growth, have been tested for their antitumor activity in tissue culture and in vivo. In this review, we describe the recent discoveries regarding the physiological properties and microbial production and metabolism of l-proline analogues, particularly l-AZC and HOPs. Their applications in fundamental research and industrial use are also discussed.     

Uptake of amino acids by actidione-treated yeast cells

The active uptake ofl-aspartic acid, glycine andl-lysine by actidione-treated cells ofSaccharomyces cerevisiae was found to be inhibited by anaerobic conditions in the absence of a source of energy, only facilitated diffusion persisting. Similarly, metabolic inhibitors (iodoacetamide, sodium fluoride and potassium sorbate) inhibited the uptake very substantially. 2,4-Dinitrophenol and sodium azide appeared to inhibit the movement of the transport carrier itself, while uranyl ions showed a complex interaction pattern, ranging from inhibition at concentrations of 10^?6–10^?4 m, to stimulation at concentrations of 3×10^?4–10^?3 m, to pronounced inhibition at higher concentrations. The uptake was pH-dependent with optima forl-aspartic acid near pH 4, for glycine near pH 5, forl-lysine near pH 6.5.     

LignicolousHyphomycetes from Czechoslovakia 3.Sporoschisma,Sporoschismopsis andCatenularia

The species of three genera—Sporoschisma Berk. etBr.,Sporoschismopsis Hol.-Jech. etHennebert andCatenularia Grove are treated in this paper from Czechoslovakia:Sporoschisma juvenile Boudier,S. mirabile Berk. etBr.,Sporoschismopsis moravica Hol.-Jech. etHennebert andCatenularia cuneiformis (Richon) Mason. The studied fungi produce the conidia in basipetal succession, endogenously or subendogenously, quite internally in large phialides or only inside open swollen collarettes.     

Properties of the sugar carrier in Baker's yeast

Incubation ofSaccharomyces cerevisiae cells withd-galactose induced the formation of galactose-utilizing enzymes, among them a monosaccharide carrier, apparently synthesized as a proteinde novo. The synthesis of the carrier preceded that of galactokinase by as much as 2 h. The inducible carrier shows a preference for monosaccharides with an axial hydroxyl group at carbon 4 of theC1 chair conformation or at carbon 2 of the1C chair conformation. Through its mediation, some sugars normally poorly transported (d-galactose,d-fucose,l-xylose,l-arabinose) can enter into the entire cell water, occupying then one more kinetic (and morphological ?) compartment than before induction. Some other monosaccharides, readily transported even by a constitutive carrier system (e.g.l-sorbose,d-xylose,d-arabinose) share the newly synthesized carrier.     

Notes on some African hepatic genera 1–5

The present study deals with five genera of hepatics in Africa, Isotachis Mitt., Anastrophyllum (Spruce) Steph., Tritomaria Schiffn. ex Loeske, Gymnocoleopsis (Schust.) Schust. and Lophozia (Dum.) Dum. All African populations of the genus Isotachis Mitt. are considered to be one species, I. aubertii (Schwaegr.) Mitt. Four species of Anastrophyllum (Spruce) Steph. (s.l.), A. auritum (Lehm.) Steph., A. piligerum (Nees) Spruce, A. subcomplicatum (Lehm. et Lindenb.) Steph. and A. minutum (Schreb.) Schust., and two species of Tritomaria Schiffn. et Loeske, T. camerunensis S. Arnell and T. exsecta (Schrad.) Schiffn. ex Loeske occur in Africa. Gymmocoleopsis multiflora (Steph.) Schust. represents a genus and species hitherto unreported for the African flora. Finally, five Lophozia (Dum.) Dum. species, L. argentina (Steph.) Schust., L. capensis S. Arnell, L. decolorans (Limpr.) Steph., L. hedbergii S. Arnell and L. tristaniana (S. Arnell) Váňa, are reported from central and southern Africa; two of these (L. argentina (Steph.) Schust. and L. decolorans (Limpr.) Steph.) represent the first reports from Africa.     

The Synthesis of l-dopa-l-Tyr and The Interaction of l-dopa-l-Tyr With ctDNA

l-dopa-l-Tyr was synthesized by Fmoc solid-phase peptide synthesis, purified by reversed-phase HPLC and characterized by using ¹H, ¹³C NMR and ESI–MS analyses. The interaction of l-dopa-l-Tyr and l-dopa with ctDNA has been investigated respectively by UV–vis absorption and fluorescence spectroscopy. The results showed that both l-dopa and l-dopa-l-Tyr interacted with ctDNA through intercalative mode and l-dopa-l-Tyr showed a higher affinity for DNA. Meanwhile, compared with the free l-dopa, gel electrophoresis assay also demonstrated that l-dopa-l-Tyr interacted with DNA by intercalation.     

Simultaneous electrochemical determination of l-cysteine and l-cysteine disulfide at carbon ionic liquid electrode

A linear sweep voltammetric method is used for direct simultaneous determination of l-cysteine and l-cysteine disulfide (cystine) based on carbon ionic liquid electrode. With carbon ionic liquid electrode as a high performance electrode, two oxidation peaks for l-cysteine (0.62 V) and l-cysteine disulfide (1.3 V) were observed with a significant separation of about 680 mV (vs. Ag/AgCl) in phosphate buffer solution (pH 6.0). The linear ranges were obtained as 1.0–450 and 5.0–700 μM and detection limits were estimated to be 0.298 and 4.258 μM for l-cysteine and l-cysteine disulfide, respectively. This composite electrode was applied for simultaneous determination of l-cysteine and l-cysteine disulfide in two real samples, artificial urine and nutrient broth. Satisfactory results were obtained which clearly indicate the applicability of the proposed electrode for simultaneous determination of these compounds in complex matrices.     

Valuation, appraisal, discounting, obsolescence and depreciation

Previous editions of this Journal have drawn attention to the critical role valuation plays in life cycle analysis and environmental impact assessment (see for exampleVolkwkin andKlöpffer |1|). In particular, the critical role of valuation has heen highlighed in a number of discussions on ‘valuation step’ within life cycle costing, ‘hedonic and contingency’ assessments of environmental impact and both the utility and welfare of ‘pathway’ analysis/assessment (Krkwitt, Mayerhofer, Trukenmüller andFriedrich, 1998;Powell, Pearce andCraighill, 1997;Volkwein, On in andKlöpffer, 1996 |2-4|). Focusing on the utility of market valuation, this paper examines the critique of discounting environmentalists have made in relation to property valuation, investment appraisal and the application of the principle in the income based net annual return model of land use time-horizons and the spatial configuration of building programmes-a criticism implict in ‘valuation step’, ‘hedonic, contingency’ and ‘pathway’ analysis/assessments. It examines the argument put forward regarding the link between the selection of a discount rate, the valuation of property, appraisal of investment and inter-generational downloading of costs associated with the use of land, repair, maintenance and refurbishment of buildings: the downloading of costs, seen by some, to have an adverse impact and work against the introduction of experimental designs aimed at energy saving, clean air environments.     

LignicolousHyphomycetes from Czechoslovakia 5.Septonema,Hormiactella andLylea

Three genera of lignicolousHyphomycetes Septonema Corda,Hormiactella Saac. andLylea Morgan-Jones are discussed. Illustrations and determination keys are provided. Five species ofSeptonema—S. fasciculare (Corda) Hughes,S. leptaleum (Ellis & Harkn.) Hughes,S. pinicola Hol.-Jech.,S. secedens Corda andS. laricium Hol.-Jech.—and two species ofHormiactella—H. fusca (Preuss) Sacc. andH. asetosa Hol.-Jech.—have been found in Czechoslovakia. A new speciesS. pseudobinum Hol.-Jech. is described from Romania.Septonema tetracoilum (Corda Hughes is accommodated in the genusLylea Morgan-Jones.     

The trilobiteProtostygina and the composition of the Styginidae, with two new genera

The Styginidae is regarded as an exclusively Ordovician family of trilobites, separate from the Scutelluidae. The hitherto poorly known genusProtostygina Prantl &P?ibyl, 1949 is revised. It is recorded with certainty only from the Llanvirn of the Czech Republic, and the type species is a senior synonym of“Raymondaspis” rubensi rubensi P?ibyl &VANěK, 1968 and“R.” rubensi lybar ?najdr, 1976. Two new styginid genera are proposed:Cyrtocybe, with type species“Raymondaspis” turgida Whittington, 1965, is known from the upper Arenig and lower Llanvirn of Newfoundland, Maine and Norway; andPromargo, with type speciesP.forteyi n. sp., occurs in the Arenig of Newfoundland and Spitsbergen.Turgicephalus Fortey, 1980 is regarded as a junior synonym ofRaymondaspis P?ibyl inPrantl &P?ibyl, 1949. Three genera are excluded from the Styginidae:Kirkdomina Tripp, 1962,Pseudostygina Zhou inZhou et al., 1982 andStyginella P?ibyl &Vaněk, 1971.     

Biosynthesis of ethylene glycol in Escherichia coli

Ethylene glycol (EG) is an important platform chemical with steadily expanding global demand. Its commercial production is currently limited to fossil resources; no biosynthesis route has been delineated. Herein, a biosynthesis route for EG production from d-xylose is reported. This route consists of four steps: d-xylose?→?d-xylonate?→?2-dehydro-3-deoxy-d-pentonate?→?glycoaldehyde?→?EG. Respective enzymes, d-xylose dehydrogenase, d-xylonate dehydratase, 2-dehydro-3-deoxy-d-pentonate aldolase, and glycoaldehyde reductase, were assembled. The route was implemented in a metabolically engineered Escherichia coli, in which the d-xylose?→?d-xylulose reaction was prevented by disrupting the d-xylose isomerase gene. The most efficient construct produced 11.7 g?L^?1 of EG from 40.0 g?L^?1 of d-xylose. Glycolate is a carbon-competing by-product during EG production in E. coli; blockage of glycoaldehyde?→?glycolate reaction was also performed by disrupting the gene encoding aldehyde dehydrogenase, but from this approach, EG productivity was not improved but rather led to d-xylonate accumulation. To channel more carbon flux towards EG than the glycolate pathway, further systematic metabolic engineering and fermentation optimization studies are still required to improve EG productivity.