Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
DNase I requires Ca²+ and Mg²+ for hydrolyzing double-stranded DNA. However, the number and the location of DNase I ion-binding sites remain unclear, as well as the role of these counter-ions. Using molecular dynamics simulations, we show that bovine pancreatic (bp) DNase I contains four ion-binding pockets. Two of them strongly bind Ca²+ while the other two sites coordinate Mg²+. These theoretical results are strongly supported by revisiting crystallographic structures that contain bpDNase I. One Ca²+ stabilizes the functional DNase I structure. The presence of Mg²+ in close vicinity to the catalytic pocket of bpDNase I reinforces the idea of a cation-assisted hydrolytic mechanism. Importantly, Poisson-Boltzmann-type electrostatic potential calculations demonstrate that the divalent cations collectively control the electrostatic fit between bpDNase I and DNA. These results improve our understanding of the essential role of cations in the biological function of bpDNase I. The high degree of conservation of the amino acids involved in the identified cation-binding sites across DNase I and DNase I-like proteins from various species suggests that our findings generally apply to all DNase I-DNA interactions.
ISSN: 1553-734X

hal-00645165
https://hal.archives-ouvertes.fr/hal-00645165
https://hal.archives-ouvertes.fr/hal-00645165/document
https://hal.archives-ouvertes.fr/hal-00645165/file/journal.pcbi.1001000.pdf
DOI : 10.1371/journal.pcbi.1001000

Éditeur(s) : Gauthier-Villars
Résumé : Five particle traps were deployed at depths of 4 400 to 4 900 rn at two stations on the Demerara abyssal plain and one station on the Cape Verde abyssal plain (tropical Atlantic). At the same time, an intensive sampling of superficial sediments was carried out using a 0,25 m2 box corer. The total particle flux varied from 372 to 87,7 mg dry weightfm2/day. The average concentration of organic carbon was 40 mg/g of material collected. ln the surface sediment this was only 4 mg/g. A direct relationship existed between the intensity of the flux of organic matter and the richness of the organic matter of the deposited sediment. According to our calculations, more than 90% of the organic matter arriving at the bottom is consumed by the abyssal benthos and only 1% is lost to permanent sediment deposits. For 100 calories consumed by the benthos, 99 are utilized by the infauna and flora, 0, 7 by the holothurian egafauna and 0,3 by near-bottom fisches. The biochemical composition of the particles is characterized at ali stations by a predominance of "humic" material in the total organic matter (more than 55%). The part of the molecules unaltered form living material varied from on station to another: for the most oceanic station the protein dominated, composing 20% of the total energy. At the station onder the influence of the continent the lipid fraction grew and was almost equal to that of the protein. ln this case, the carbohydrate fraction was very weak (7% of the organic matter). Finally the preference of the animais which feed on the particles is principally for the most energetic (lipid) and the most easily hydrolysable (labile protein) molecules: the other molecules, such as the humic matter, are less preferred, although they still contribute to about half of the energy consumed by the abyssal benthic animais.
Cinq pièges à particules ont été déployés à des profondeurs comprises entre 4400 et 4 900 rn sur deux stations dans la plaine abyssale de Demerara et une station dans la plaine abyssale du Cap Vert (Atlantique tropical). Dans le même temps, un échantillonnage intensif du sédiment superficiel a été réalisé à l'aide d'un carottier de 0,25 m2 d'ouverture. Le flux particulaire total mesuré varie de 372 mg de matière sèche/m2/jour à 87,7 mg/m2/g de matériel récolté. Dans les sédiments superficiels, elle n'est plus que d'environ 4 mg/g. Il existe une relation directe entre l'intensité du flux organique et la richesse en composés organiques du sédiment. D'après nos estimations, plus de 90% du matériel organique arrivant sur le fond sont consommés par les organismes benthiques abyssaux et seulement 1% est perdu dans la sédimentation permanente. Pour 100 calories consommées par le benthos, 99 le sont par la petite faune et la flore vivant dans le sédiment, 0, 7 calories sont utilisées par les holothuries (mégafaune) et 0,3 calories par les poissons. La composition biochimique des particules se caractérise, à toutes les stations, par la prédominance des composés organiques transformées néobiogéniques ( « humus ») dans la matière organique totale (plus de 55%). La part des molécules de la matière vivante varie d'une station à l'autre: à la station la plus océanique, les protéines dominent et composent 200/o de l'énergie totale des apports. Dans la station plus soumise aux influences continentales, la concentration en lipides augmente et peut égaler celle des protéines. Dans ce cas, le taux de glucides des particules est très faible (7% des apports organiques). Enfin, les préférences nutritionnelles des animaux qui s'alimentent sur ces particules se portent principalement sur les molécules les plus énergétiques (lipides) et les plus facilement hydrolysables (protéines labiles) : les autres molécules, comme les composés néobiogéniques sont moins recherchés bien qu'ils contribuent pour moitié à l'énergie consommée par les animaux benthiques abyssaux
Oceanologica Acta (0399-1784) (Gauthier-Villars), 1985 , Vol. 8 , N. 3 , P. 293-301

Droits : Gauthier-Villars
http://archimer.ifremer.fr/doc/00112/22318/19991.pdf
http://archimer.ifremer.fr/doc/00112/22318/

Résumé : Bacteriological study of abyssal sediments: During the Demeraby mission of the N/O Jean Charcot in the western Atlantic Ocean, a study of the microflora of the digestive tracts of echinoderms (Holothuroidea of various genera, Asteroidea, Echinoidea) was compared with that of the surrounding sediment. The bacterial concentration observed in the digestive tracts of all the animals studied is higher than that in the surrounding sediments. The largest numbers were always found in the most anterior part of the digestive tracts of the holothurians. In some cases, the increase is considerable (7.10 versus 5.2. 10 cells per millilitre): It indicates a proliferation of bacterial cells in the anterior part of the digestive tract. As the sediment moves through the digestive tract, the bacterial level progressively decreases. In the most posterior part, the concentration is, however, always clearly greater than that in the surrounding sediment. In the case of the holothurian of the genus Deima, analysis according to the Adamsonian principle of comparison shows that the microflora isolated from the various parts of the digestive tract is composed of taxonomic groups showing little resemblance to those isolated from the sediment. In the case of the holothurian of the genus Pseudostichopus, no specialisation of the microflora from digestive tract contents appears with respect to that of the ingested sediment. Examination of metabolic activities suggests a juxtaposition of two types of communities: Free sediment - sediment in transit that complement one other in their enzymatic and nutritional behaviour Indeed, the strains of free sediment seem to be rather suitable for hydrolysing and using carbohydrates, while those of enteric communities prefer the hydrolysis of lipids and proteins and the assimilation of fatty acids, dicarboxylic acids, hydroxylic acids and certain amino acids. Moreover, the enteric microflorae possess a greater nutritional versatility that, linked with its total numbers, has a sizeable catabolic potential. .../... -2- Measurements of heterotrophic activity done by means of 14 C glucose tracers make it possible to show that the metabolic activities of the bacterial communities of superficial abyssal sediments vary according to zones. In the Amazon Cone, a clear glucose mineralization gradient over a distance of a few kilometres can be observed. In the abyssal plain of Demerara heterotrophic activities are slowed in relation to those of the superficial sediments of the Cone of the Amazon, even though the bacterial numbers and the depths are of the same order of magnitude in the two zones. The abundant microflorae associated with the various divisions of digestive tract contents of the Deima abyssal holothurian manifest little metabolic activity as regards the glucose. However, the bacterial communities of Pseudostichopus and Psychropotes manifest very strong heterotrophic activities, increasing from the oesophagus towards the cloaca. Current results are too fragmentary to allow us to observe possible physiological specialisation of enteric bacterial communities enabling the facilitation of digestion processes in the host. Conclusion: In the abyssal environment, characterised by poorly developed bacterial communities, microflorae associated with the digestive tract contents of echinoderms have considerably greater cellular concentrations. By means of their enzymatic equipment, these enteric communities, which complement those of the free microflorae of superficial sediments, can participate in the transformation of organic matter in the abyssal area. Study of the dynamics of microbial communities and their heterotrophic activities in shellfish farms: From experiments done on the water in the Arcachon basin, trials to quantify and describe the reactivity potential of bacterial communities were performed. Bacterial communities were defined by direct counts, counts after setting them up for cultivation, structure and diversity of these communities, dosage of AMP, ADP, ATP. Two opposite ecological conditions were considered: Autumn conditions (high temperature) and a winter conditions (low temperature). Tubs of sea water of 6 and 60 litres were enriched either with ammonium chloride or a mixture of amino acids, at about 500 uAtg N.L. The experiments lasted 24 hours, with sampling every 3 hours. The autumn community reacted rapidly (6 hours), the response to the enrichment in organic nitrogen being the highest. The structure of the community has unstable immature stages. On the contrary, during the winter experiment, no response could be perceived. This may mean that the latency phase is greater than 24 hours. The structure of this community has mature stages. In conclusion, bacterial communities react in the same way as macroscopic communities. Stable communities are diversified and correspond to mature stages of the ecosystem; the immature stages are unstable and less diversified. This type of experiment has made it possible to bring to light the rapidity of response of bacterial populations, and can be used to define the stability of a bacterial community in a precise biotope.
ETUDE BACTERIOLOGIQUE DES SEDIMENTS ABYSSAUX RESUME Au cours de la mission DEMERABY du n/o Jean CHARCOT en atlantique occidental , une étude de la microflore des tractus digestifs d'Echinodermes (Holothurides de différents genres, Astérides, Echinides) a été comparée à celle du sédiment environnant. La concentration bactérienne observée dans les tractus digestifs de tous les animaux étudiés est supérieure à celle des sédiments environnants. Les effectifs les plus importants sont toujours observés dans la partie la plus antérieure des tractus digestifs des holothuries. Dans certains cas, l'accroissement est considérable (7.10 contre 5,2. 10 cellules/ml) : il indique une prolifération des cellules bactériennes dans la partie antérieure du tractus digestif. Au.cours de la progression du sédiment dans le tractus digestif, la charge bactérienne décroit progressivement. Dans la partie la plus postérieure, la concentration est cependant toujours nettement supérieure à celle du sédiment environnant-. Dans le cas d'une holothurie du genre Deima, l'analyse selon le principe adansonien de comparaison montre que la microflore isolée des différentes parties du tractus digestif est constituée de groupes taxonomiques présentant peu de ressemblance avec ceux isolés du sédiment. Dans le cas d'une holothurie du genre Pseudostichopus il n'apparait pas de spécialisation de la microflore du contenu du tractus digestif par rapport à celle du sédiment ingéré. ¿ ¿ . L'étude des activités métaboliques suggère une juxtaposition de deux types de communautés : sédiment libre-sédiment en transit qui se complètent dans leur comportement enzymatique et nutritionnel. En effet les souches du sédiment libre semblent être plutôt aptes à hydrolyser et à utiliser les glucides, tandis que celles des communautés entériques portent leur préférence sur l'hydrolyse des lipides et protéines et 1'assimilation des acides gras, acides dicarboxyl iques, hydro'xy-acides et certains acides aminés. D'autre part les microflores entériques possèdent une versatilité nutritionnelle plus importante qui, liée à leurs effectifs, offre une potentialité catabolique non négligeable. .../... -2- Les mesures d'activité hétérotrophe effectuées au moyen de traceurs glucose 14 C permettent de montrer que les activités métaboliques des communautés bactériennes des sédiments abyssaux superficiels varient selon les zones. Dans le Cone de l'Amazone on observe un net gradient de minéralisation du glucose sur une distance de quelques kilomètres. Dans la plaine abyssale de Demerara les activités hétérotrophessont ralenties par rapport à celles des sédiments superficiels du Cone de l'Amazone, bien que les effectifs bactériens et les profondeurs soient du même ordre de grandeur dans ces deux zones. Les abondantes microflores associées aux différentes fractions du contenu du tractus digestif de l'holothurie abyssale Deima ne manifestent que peu d'activités métaboliques vis à vis du glucose. Par contre les communautés . bactériennes de Pseudostichopus et P syahropotes manifestent des activités hétérotrophes très fortes, croissant de 1'oesophage vers le cloaque. Les résultats actuels sont trop fragmentaires pour permettre d'observer une éventuelle spécialisation physiologique des communautés bactériennes entëriques permettant de faciliter les processus de digestion chez l'hôte. Conclusion: dans le milieu abyssal, caractérisé par des communautés bactériennes faiblement développées, les microflores associées aux contenus de tractus digestif des Echinodermes présentent des concentrations cellulaires considérablement plus importantes. Par leurs équipements enzymâtiques ces communautés entëriques complémentaires de ceux des microflores libres des sédiments superficiels, peuvent participer à la transformation des matières organiques du domaine abyssal. ETUDE DE LA DYNAMIQUE DES COMMUNAUTES MICROBIENNES ET DE LEURS ACTIVITES HETEROTROPHES DANS LES PARCS CONCHYLICOLES RESUME A partir d'expérimentations effectuées sur l'eau du bassin d'Arcachon, des essais de quantification et qualification du potentiel de réactivité des communautés bactériennes ont été effectuées. Les communautés bactériennes ont été définies par comptage directs, comptages après mise en culture, structure et diversité de ces communautés, dosage des AMP, ADP, ATP. Deux situations écologiques opposées ont étées considérées . Une situation automnale (température élevée) et une situation hivernale (température faible). Des bacs d'eau de mer de 6 et 60 litres ont été enrichis , soit avec du chlorure d'ammonium, soit avec un mélange d'acides aminés, à environ 500 uAtg N.l" . Les expériences ont duré 24 heures, avec des prélèvements toutes les 3 heures. La communauté automnale réagit rapidement (6 heures), la ' réponse à l'enrichissement en azote organique étant la plus importante. La structure de la communauté présente des stades immatures instables. Au contraire, au cours de l'expérimentation hivernale, aucune réponse n'a pu être perçue. Ceci peut signifier que la phase de latence est supérieure à 24 heures. La structure . de cette communauté présente des stades matures. En Conclusion, les communautés bactériennes réagissent de la même façon que les communautés macroscopiques. Les communautés stables sont diversifiées et correspondent à des stades matures de l'écosystème, les stades immatures sont instables et moins diversifiés. Ce type d'expérience a permi de mettre en évidence la rapidité de réponse des populations bactériennes, et peut être utilisé pour définir la stabilité d'une communauté bactérienne dans un biotope précis.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/1981/rapport-1926.pdf
http://archimer.ifremer.fr/doc/00000/1926/

Éditeur(s) : Elsevier
Résumé : Until recently, it was considered impossible to feed newly hatched marine fish species with a compound diet. Substituting a compound diet for live prey was performed several weeks after hatching, depending on the species. Compound diets were well ingested at the early stage but larvae died with a gut full of food, suggesting that larvae were unable to digest the compound diet. The hypothesis was that younger larvae have insufficient digestive enzymes to thrive on compound diets, and that exogenous enzymes, provided from live prey, are necessary for early stages.The organogenesis of marine fish larvae is not completely achieved at hatching and histological studies have revealed that the anatomy of the digestive tract undergoes developmental changes over some weeks. Nevertheless, biochemical studies over 20 years have shown that most of the digestive enzymes are present in young larvae. Recent studies have provided better understanding of digestion mechanisms in larvae and have led to proposed dietary compositions meeting larvae nutritional requirements. Pancreatic digestive enzymes are detected before mouth opening. Their synthesis is not induced by diet ingestion, but secretory mechanisms in the pancreas, and so enzymatic action, become efficient chronologically after those of synthesis. Inadequate diets can delay the onset of secretion mechanisms. The ratio of secreted enzymes to total enzymes indicates the nutritional value of the diet ingested by the larvae.At the intestinal level, cytosolic enzymes, which are peptidases, exhibit high activity in the early stages, suggesting a high capacity in larvae to digest protein hydrolysate. Indeed, larvae growth and survival is improved by the incorporation of a moderate concentration of peptide or hydrolysate in the diet. Peptidase activity abruptly decreases around day 25 in sea bass, concurrent with an increase in enzymes of the brush border membranes. This corresponds to a normal maturation process of enterocytes. Compound diets can slightly delay the onset of this maturation process, and inadequate diet can prevent it, leading to near death of the larvae. A proper onset of the maturation process has been associated with high larvae survival.The early developmental stage larvae exhibit high hydrolytic capacity, related to their weight. Enzyme activity pattern is age-dependent, but can be modulated by diet composition. Thus, larvae have the ability to digest and thrive on compound diet, if this diet is well adapted. Larvae have different specificities in digestion and nutritional requirements when compared to juveniles. Taking these specificities into consideration, recent research has led to the formulation of a compound diet that was well adapted for larvae from mouth opening, and could totally replace live prey.
Aquaculture (0044-8486) (Elsevier), 2001-08 , Vol. 200 , N. 1-2 , P. 161-180

Droits : 2001 Elsevier Science B.V. All rights reserved
http://archimer.ifremer.fr/doc/2001/publication-447.pdf
DOI:10.1016/S0044-8486(01)00699-8
http://archimer.ifremer.fr/doc/00000/447/

Éditeur(s) : HAL CCSD BioMed Central
Résumé : International audience
Background : Trichoderma reesei is the main industrial source of cellulases and hemicellulases required for the hydrolysis of biomass to simple sugars, which can then be used in the production of biofuels and biorefineries. The highly productive strains in use today were generated by classical mutagenesis. As byproducts of this procedure, mutants were generated that turned out to be unable to produce cellulases. In order to identify the mutations responsible for this inability, we sequenced the genome of one of these strains, QM9136, and compared it to that of its progenitor T. reesei QM6a.Results : In QM9136, we detected a surprisingly low number of mutagenic events in the promoter and coding regions of genes, i.e. only eight indels and six single nucleotide variants. One of these indels led to a frame-shift in the Zn2Cys6 transcription factor XYR1, the general regulator of cellulase and xylanase expression, and resulted in its C-terminal truncation by 140 amino acids. Retransformation of strain QM9136 with the wild-type xyr1 allele fully recovered the ability to produce cellulases, and is thus the reason for the cellulase-negative phenotype. Introduction of an engineered xyr1 allele containing the truncating point mutation into the moderate producer T. reesei QM9414 rendered this strain also cellulase-negative. The correspondingly truncated XYR1 protein was still able to enter the nucleus, but failed to be expressed over the basal constitutive level.Conclusion : The missing 140 C-terminal amino acids of XYR1 are therefore responsible for its previously observed auto-regulation which is essential for cellulases to be expressed. Our data present a working example of the use of genome sequencing leading to a functional explanation of the QM9136 cellulase-negative phenotype.
ISSN: 1471-2164

Droits : http://creativecommons.org/licenses/by/
hal-01149671
http://hal.upmc.fr/hal-01149671
http://hal.upmc.fr/hal-01149671/document
http://hal.upmc.fr/hal-01149671/file/Lichius_2015_Genome_sequencing.pdf
DOI : 10.1186/s12864-015-1526-0