Continuous enzymatic transformation in an enzyme membrane reactor with simultaneous NAD(H) regeneration

Multienzyme reaction systems with simultaneous coenzyme regeneration have been investigated in a continuously operated membrane reactor at bench scale. NAD(H) covalently bound to polyethylene glycol with a molecular weight of 10⁴ [PEG-10,000-NAD(H)] was used as coenzyme. It could be retained in the membrane reactor together with the enzymes. L -leucine dehydrogenase (LEUDH) was used as catalyze for the reductive amination of α-ketoisocaproate (2-oxo-4-methylpentanoic acid) to L -leucine. Format dehydrogenease (FDH) was used for the regeneration of NADH. Kinetic experiments were carried out to obtain data which could be used in a kinetic model in order to predict the performance of an enzyme membrane reactor for the continuous production of L -leucine. The kinetic constants V_max and K_m of enzymes are all in the same range regardless of whether native NAD(H) or PEG-10,000-NAD(H) is used as coenzyme. L -leucine was produced continuously out of α-ketoisocaproate for 48 days; a maximal conversion of 99.7% was reached. The space-time yield was 324 mmol/L day (or 42.5 g/L day).     

Na movements and their oscillations during fertilization and the cell cycle in sea urchin eggs

We have analysed in detail the Na⁺ content and Na⁺ influx during fertilization and first divisions of the sea urchin egg (Paracentrotus lividus) using a filtration technique devised to eliminate rapidly contamination by the Na⁺ of external sea water. In the first 5 min following fertilization the egg fills up with Na⁺ (+ 30%). Thereafter Na⁺ is extruded and the Na⁺ content stabilizes at about 60% of the unfertilized egg level by the second cleavage (2 h). The initial increase in Na⁺ content is due to a large increase in Na⁺ influx already detected at 20 sec. The Na⁺ influx reaches its maximum at 1 min and its minimum at 5 min. H⁺ excretion follows the same kinetics. A second increase in Na⁺ influx is noted 5–10 min after fertilization; it reaches its maximum at prophase metaphase (30 min) and its minimum during cleavage (60 min). These oscillations in Na⁺ influx were observed for the first three divisions. Fertilization also immediately stimulates the Na⁺ efflux which remains elevated throughout the cell cycle and is responsible for the depletion of the Na⁺ content of the embryos. Activation of the eggs by weak amine bases (5 mM NH₄Cl) which bypasses the early cortical reaction produces only a depletion in the Na⁺ content of the egg similar to that produced by fertilization. NH₄Cl also increases the Na⁺ influx soon after fertilization, although no transient variations are noted.     

Interêts du dosage des adénosines 5′-phosphate pour l'étude de la dynamique des populations phytoplanctoniques lacustres (le Pavin - France)

A study on A.T.P., A.D.P. and A.M.P. in the phytoplankton of Lake Pavin was carried out from November 1976 to December 1977. Samples were collected at nine depths, between the surface and 50 m.A.T.P., A.D.P. and ( = A.T.P.+ A.D.P. + A.M.P.) show a similar behaviour in space and time but A.M.P. behaves independently. The A.M.P. concentration is high only when primary production of phytoplankton is high.The energy charge [CE = (ATP + % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaWaaSGaaeaaca% aIXaaabaGaaGOmaaaaaaa!3776!\[{\raise0.7ex\hbox{$1$} \!\mathord{\left/ {\vphantom {1 2}}\right.\kern-\nulldelimiterspace}\!\lower0.7ex\hbox{$2$}}\]ADP)/(ATP + ADP + AMP)] could be used as measure of adaptation in algal populations. Indeed, the energy charge increases as algal cells experience difficulty in maintaining themselves in the biotope.

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Interêts du dosage des adénosines 5-phosphate pour l'étude de la dynamique des populations phytoplanctoniques lacustres (le Pavin - France)

     

Megagyrolithes ardescensis N. Gen., N. SP.,Trace fossile nouvelle du Valanginien d'Ardeche (France)

A new trace fossil corresponding to a spiral burrow is described in the Upper Valanginian of the Ardèche area (France). It seems to be restricted to marine hemipelagic facies (outer shelf and upper slope).     

Nucleolar localization of ‘template-bound’ RNA polymerase I in nuclei of Xenopus laevis blastulae

This study shows, by means of autoradiography, that RNA polymerase I activity is present in some of the nuclei isolated from Xenopus blastulae. This activity is localized in one or two nucleoli which have a diameter of at least 1.5 μm. The ratio between nucleolar and total nuclear grain counts allows a quantitative estimation of RNA polymerase I activity relative to total RNA polymerase activity at the blastula stage.     

In vitro adhesion of tufted oral streptococci toBacterionema matruchotii

The adhesive interaction between the ectosymbionts of the corn cob configuration (CCC), a naturally occurring bacterial consortium in human dental plaque, was studied. In vitro association was produced in mixed cultures consisting ofBacterionema matruchotii and streptococci resemblingStreptococcus sanguis previously isolated from human CCC. Phase-contrast, selective immunofluorescence, and scanning electron microscopy showed that in this aggregative system, a tuftlike polar structure typical of the coccal epibiont mediated binding to the core filament. This aggregative model provides evidence that a specialized structure on the cell surface of epiphytic organisms may participate in interbacterial adherence.     

Réflexions critiques sur l'écologieet la systématique des Lingules actuelles et fossiles

New topics on ecology and systematics of recent and fossil Lingulids lead to an obvious revision of our knowledges on this zoological group. At first, the recent species need systematics and taxonomy on the bases of new described specific criteria (as, morphology of deltidial areas, muscle disposition); the results are briefly indicated. But, in fossil species, disorder and disparity of used characteristics are emphasized.The general conceiving on ecology of Lingulids are reviewed and discussed, especially on bathymetry and salinity; sediment and oxygenation conditions; taphocoenosis and lingulid «communities. On recent species, all these points are also studied, especially some ecological requirements (salinity, bathymetry, grain size), and mechanism of burrowing ability, burrow living positions in the sediments, as shell preservations after death and fossilization, to facilitate the paleobiotope interpretations. Recent animals are euryhalin, surviving at salinities from about 13 to 42‰; they could be considered as well adapted to waters with strong salinity fluctuations. They show preference to fine sand bottoms (lowest particle size about 40–60 μm). Their bathymetric distribution occurs between 0 and about 500 m (Lingula especially between 5–50 m; Glottidia 15–70 m). The isotherms 8–10°C seem to restrict their geographical and bathymetric distribution.Therefore, some post-palaeozoic lingulid bedsare studied or redescribed on the bases of the above discussed characteristics, and new interpretations on the environmental situation are given (Trias of Vosges Mountains; Oligocene from Japan; Eocene of London Basin). More caution must be used in study of fossil Lingulids that are not especially animals living in infralittoral bottoms with low salinity and deficient oxygenation, as generally accepted.     

Regulation of adenosine-5′-phosphosulfate sulfotransferase activity by H2S in Lemna minor L.

When Lemna minor L. is transferred to an atmosphere with H₂S, there is a rapid loss of extractable adenosine-5-phosphosulfate sulfotransferase activity. The activity is restored within 24 h in an atmosphere without H₂S. This restoration of activity is completely inhibited by cycloheximid but not by chloramphenicol. In vitro, S^2- up to 5 mM and cysteine, methionine, and glutathione up to 50 mM do not inhibit the enzyme. The activities of ATP sulfurylase and O-acetyl-L-serine sulfhydrylase are not affected significantly by H₂S. The physiological significance of the regulation of adenosine-5-phosphosulfate sulfotransferase is discussed.Abbreviations APS adenosine-5-phosphosulfate - PAPS adenosine-3-phosphate-5-phosphosulfate - BSA bovine serum albumin - DTT dithiothreitol - POPOP 1,4-di [2-(5-phenyloxazolyl)]-benzene - PPO 2,5-diphenyloxazol This is no. 6 in the series Regulation of Sulfate Assmilation in Plants