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Registros recuperados: 8
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A novel lipase / acyltransferase from the yeast Candida albicans: expression and characterisation of the recombinant enzyme Inra
Roustan, J.L.; Rascon Chu, A.; Moulin, G.; Bigey, F..
A gene encoding an extracellular lipase (CaLIP4) from Candida albicans was successfully expressed in Saccharomyces cerevisiae after mutagenesis of its unusual CUG serine codon into a universal one. The ability of this lipase, which shares 60% sequence homology with the lipase/acyltransferase from Candida parapsilosis, to synthesise esters was investigated. CaLIP4 behaved as a true lipase, displaying activity towards insoluble triglycerides and having no activity in the presence of short-chain fatty acid (FA) esters and phosphatidylcholine. Methyl, ethyl and propyl esters were efficiently used. The lipase exhibited highest selectivity for unsaturated FA. With saturated FAs, C14-C16 acyl chains were preferred. In a biphasic aqueous/lipid system, CaLIP4...
Tipo: Journal Article Palavras-chave: CANDIDA ALBICANS; CANDIDA PARAPSILOSIS; SACCHAROMYCES CEREVISIAE; ALCOHOLYSIS ACTIVITY; LIPASE/ACYLTRANSFERASE; KINETIC CONTROL.
Ano: 2005 URL: http://www.prodinra.inra.fr/prodinra/pinra/doc.xsp?id=PROD20084762bbfc&uri=/notices/prodinra1/2010/09/
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Clusterin increases post-ischemic damages in organotypic hippocampal slice cultures Inra
Hakkoum, D.; Imhof, A.; Vallet, P.G.; Boze, H.; Moulin, G.; Charnay, Y.; Stoppini, L.; Aranow, B.; Bouras, C.; Giannakopoulos, P..
Clusterin or apolipoprotein J is a heterodimeric glycoprotein which is known to be increased during tissue involution in response to hormonal changes or injury and under circumstances leading to apoptosis. Previous studies in wild-type (WT) and clusterin-null (Clu-/-) mice indicated a protective role of clusterin over-expression in astrocytes lasting up to 90 days post-ischemia. However, in in vitro and in vivo models of neonatal hypoxia-ischemia, clusterin exacerbates necrotic cell death. We developed recombinant forms of clusterin and examined their effect on propidium iodide uptake, neuronal and synaptic markers as well as electrophysiological recordings in hippocampal slice cultures from Clu-/- and WT mice subjected to oxygen-glucose deprivation (OGD)....
Tipo: Journal Article Palavras-chave: CLUSTERIN; GLUTAMATE RECEPTOR; MULTI-ELECTRODE ARRAY; ORGANOTYPIC HIPPOCAMPAL SILICE CULTURE; OXYGENE-GLUCOSE DEPRIVATION.
Ano: 2008 URL: http://www.prodinra.inra.fr/prodinra/pinra/doc.xsp?id=PROD2010f9a5b795&uri=/notices/prodinra1/2010/09/
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Complete hydrolysis of myo-inositol hexakisphosphate by a novel phytase from Debaryomyces castellii CBS 2923 Inra
Ragon, M.; Aumelas, A.; Chemardin, P.; Galvez, S.; Moulin, G.; Boze, H..
Debaryomyces castellii phytase was purified to homogeneity in a single step by hydrophobic interaction chromatography. Its molecular mass is 74 kDa with 28.8% glycosylation. Its activity was optimal at 60 C and pH 4.0. The Km value for sodium phytate was 0.532 mM. The enzyme exhibited a low specificity and hydrolyzed many phosphate esters. The phytase fully hydrolyzed myo-inositol hexakisphosphate ( or phytic acid, Ins P-6) to inositol and inorganic phosphate. The sequence of Ins P-6 hydrolysis was determined by combining results from high-performance ionic chromatography and nuclear magnetic resonance. D. castellii phytase is a 3-phytase that sequentially releases phosphate groups through Ins ( 1,2,4,5,6) P-5, Ins ( 1,2,5,6) P-4, Ins ( 1,2,6) P-3, Ins (...
Tipo: Journal Article Palavras-chave: DEBAROMYCES CASTELLII; YEAST; PHYTASE; ENZYME CHARACTERIZATION; INOSITOL PHOSPHATE; PHYTATE DEGRADATION; NMR; HPLC.
Ano: 2008 URL: http://www.prodinra.inra.fr/prodinra/pinra/doc.xsp?id=PROD2009150d6498&uri=/notices/prodinra1/2010/09/
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Inhibition of Clostridium butyricum by 1,3-propanediol and diols during glycerol fermentation Inra
Colin, T.; Bories, A.; Moulin, G..
1,3-Propanediol inhibition during glycerol fermentation to 1,3-propanediol by Clostridium butyricum CNCM 1211 has been studied. The initial concentration of the 1,3-propanediol affected the growth of the bacterium more than the glycerol fermentation. μ max was inversely proportional to the initial concentration of 1,3-propanediol (0–65 g l−1). For glycerol at 20 g l−1, the growth and fermentation were completely stopped at an initial 1,3-propanediol concentration of 65 g l−1. However, for an initial 1,3-propanediol concentration of 50 g l−1 and glycerol at 70 g l−1, the final concentration (initial and produced) of 1,3-propanediol reached 83.7 g l−1(1.1 M), with complete consumption of the glycerol. Therefore, during the fermentation, the strain tolerated...
Tipo: Journal Article Palavras-chave: CLOSTRIDIUM BUTYRICUM; PROPANEDIOL; BIOTECHNOLOGIE; PRESSION OSMOTIQUE.
Ano: 2000 URL: http://www.prodinra.inra.fr/prodinra/pinra/doc.xsp?id=PUB0300012739095456&uri=/notices/prodinra1/2010/09/
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Molecular gene cloning and overexpression of the phytase from Debaryomyees castellii CBS 2923 Inra
Ragon, M.; Neugnot-Roux, V.; Chemardin, P.; Moulin, G.; Boze, H..
The ORF encoding the Debaryomyces castellii CBS 2923 phytase was isolated. The deduced 461-amino-acid sequence corresponded to a 51.2 kDa protein and contained the consensus motif (RHGXRXP) which is conserved among phytases. No signal sequence cleavage site was detected. Nine potential N-glycosylation sites have been predicted. The protein shared 21-69% sequence identities with various phytases of yeast or fungal origin. Heterologous expression of the D. castellii CBS 2923 phytase in the methylotrophic yeast Pichia pastoris was tested under both the P. pastoris inducible alcohol oxidase (AOX1) promoter and the constitutive glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter. Maximum production levels obtained were 476 U ml(-1), with the AOX1 expression...
Tipo: Journal Article Palavras-chave: GENE ISOLATION; PHYTASE; DEBARYOMYCES CASTELLII; PICHIA PASTORIS; HETEROLOGOUS EXPRESSION.
Ano: 2008 URL: http://www.prodinra.inra.fr/prodinra/pinra/doc.xsp?id=PROD2009ea2756c4&uri=/notices/prodinra1/2010/08/
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NMR structure of rALF-Pm3, an anti-lipopolysaccharide factor from shrimp: model of the possible lipid A-binding site Inra
Yang, Y.; Boze, H.; Chemardin, P.; Padilla, A.; Moulin, G.; Tassanakajon, A.; Pugnière, M.; Roquet, F.; Destoumieux-Garzon, D.; Gueguen, Y.; Bachère, E.; Aumelas, A..
The anti-lipopolysaccharide factor ALF-Pm3 is a 98-residue protein identified in hemocytes from the black tiger shrimp Penaeus monodon. It was expressed in Pichia pastoris from the constitutive glyceraldehyde-3-phosphate dehydrogenase promoter as a folded and N-15 uniformly labeled rALF-Pm3 protein. Its 3D structure was established by NMR and consists of three alpha-helices packed against a four-stranded beta-sheet. The C-34-C-55 disulfide bond was shown to be essential for the structure stability. By using surface plasmon resonance, we demonstrated that rALF-Pm3 binds to LPS, lipid A and to OM(R)-174, a soluble analogue of lipid A. Biophysical studies of rALF-Pm3/LPS and rALF-Pm3/OM(R)-174 complexes indicated rather high molecular sized aggregates, which...
Tipo: Journal Article Palavras-chave: ANTI-LIPOPOLYSACCHARIDE FACTOR ; LIPOPOLYSACCHARIDE; LIPID A; NMR; STRUCTURE; SEPTIC SHOCK; SURFACE PLASMON RESONANCE ULTRACENTRIFUGATION.
Ano: 2009 URL: http://www.prodinra.inra.fr/prodinra/pinra/doc.xsp?id=PROD201064bb8a78&uri=/notices/prodinra1/2010/09/
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Overexpression and characterization of two human salivary proline rich proteins Inra
Pascal, C.; Bigey, F.; Ratomahenina, R.; Boze, H.; Moulin, G.; Manchado-Sarni, P..
Proline rich proteins (PRP) are among major human saliva constituents and are known to interact with wine tannins that are involved in astringency. To characterize these interactions, a human salivary proline rich pro-protein, PRB4S, was overexpressed in Pichia pastoris. Six recombinant proteins resulting from maturation in bioreactor were detected by SDS-PAGE analysis between 15 and 45 kDa (apparent molecular weight). Two of them, the 45 and the 15 kDa ones, were isolated from culture supernatant by adsorption and permeation chromatography. They were characterized by N-terminal sequencing and MALDI-TOF analysis after trypsic digestion. The 45 kDa protein is glycosylated while the 15 kDa one was obtained after a furin-like proteolysis. Both of them are...
Tipo: Journal Article Palavras-chave: HUMAN SALIVARY PRP; PRB4S; PICHIA PASTORIS; OVEREXPRESSION; TRYPSIC DIGESTION; MALDI-TOF; N-TERMINAL SEQUENCE; INTRINSICALLY UNSTRUCTUREDPROTEIN; II-1; IB-5.
Ano: 2006 URL: http://www.prodinra.inra.fr/prodinra/pinra/doc.xsp?id=PROD200846b46bb&uri=/notices/prodinra1/2009/03/
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Recombinant expression and anti-microbial activity of anti-lipopolysaccharide factor (ALF) from the black tiger shrimp Penaeus monodon Inra
Somboonwiwat, K.; Marcos, M.; Tassanakajon, A.; Klinbunga, S.; Aumelas, A.; Romestand, B.; Gueguen, Y.; Boze, H.; Moulin, G.; Bachère, E..
Anti-lipopolysaccharide factors (ALFs), originally characterized from horseshoe crabs, have been recently identified from hemocytes of the black tiger shrimp, Penaeus monodon, by a genomic approach. In order to characterize the properties and biological activities of this immune effector in shrimp, ALFPm3, the most abundant isoform found in P. monodon, was expressed in the yeast Pichia pastoris. Large-scale production in fermentor provided 262 mg/l of recombinant ALFPm3 which was purified to homogeneity by single chromatography step on expanded-bed Streamline SP6XL. The rALFPm3 was further characterized in terms of N-terminal sequencing and mass spectrometry. Anti-microbial assays demonstrated that rALFPm3 has a broad spectrum of anti-fungal properties...
Tipo: Journal Article Palavras-chave: SHRIMP; CRUSTACEAN; ANTI-LIPOPOLYSACCHARIDE FACTOR; LPS-BINDING; ANTI6mICROBIAL ACTIVITY; VIBRIO; HEMOCYTE.
Ano: 2005 URL: http://www.prodinra.inra.fr/prodinra/pinra/doc.xsp?id=PROD20092e49f543&uri=/notices/prodinra1/2010/10/
Registros recuperados: 8
Primeira ... 1 ... Última
 

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