References of "De Pauw, Edwin"
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See detailIsolation and Cultivation of a Xylanolytic Bacillus subtilis Extracted from the Gut of the Termite Reticulitermes santonensis
Tarayre, Cédric ULg; Brognaux, Alison ULg; Brasseur, Catherine ULg et al

in Applied Biochemistry and Biotechnology (2013)

The aim of this work was the isolation of xylanolytic microorganisms from the digestive tract of the termite Reticulitermes santonensis. The reducing sugars released after the hydrolysis of xylans can be ... [more ▼]

The aim of this work was the isolation of xylanolytic microorganisms from the digestive tract of the termite Reticulitermes santonensis. The reducing sugars released after the hydrolysis of xylans can be further fermented to provide bioethanol. A xylanolytic strain of Bacillus subtilis was isolated from the hindgut of the termite and displayed amylase and xylanase activities. The bacterium was grown on media containing agricultural residues: wheat bran, wheat distiller’s grains, and rapeseed oil cake. Wheat bran led to the highest induction of xylanase activity, although the development of the strain was less fast than in the other media. It was possible to reach maximal xylanase activities of 44.3, 33.5, and 29.1 I.U./ml in the media containing wheat bran, wheat distiller’s grains, and rapeseed oil cake, respectively. Mass spectrometry identified a wide range of xylose oligomers, highlighting an endoxylanase activity. The enzyme was stable up to 45 °C and displayed an optimal pH close to 8. [less ▲]

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See detailDistribution and identification of molecular interactions between tomato roots and bacterial biofilms
Debois, Delphine ULg; Jourdan, Emmanuel; Smargiasso, Nicolas ULg et al

Poster (2013, June 12)

Some non-pathogenic microorganisms evolving in the root micro-environment can trigger a positive effect on plant, increasing host defense against disease or/and directly inhibiting growth of pathogen in ... [more ▼]

Some non-pathogenic microorganisms evolving in the root micro-environment can trigger a positive effect on plant, increasing host defense against disease or/and directly inhibiting growth of pathogen in soil (1). To initiate both phenomena leading to biocontrol activity, microorganisms use plant exudates to grow on roots and to produce in-situ active compounds. In Bacilli, cyclic lipopeptides of the surfactin, iturin and fengycin families represent important antibiotics involved in biocontrol (2). Recent studies in microbiology allowed a better understanding of plant microorganism interactions but few has been done at the molecular level. In this study, MALDI MS imaging has been used to study the nature of the secreted lipopeptide molecules, their relative quantity and their distribution in the root’s environment.Disinfected tomato seeds were first germinated at 28°C in sterile conditions for germination. Seedlings were then placed in Petri dish on ITO glass slide recovered with a thin layer of plant nutritive solution containing 1,75% of agar and treated with freshly-grown cells of Bacillus amyloliquefaciens S499. Petri dishes were incubated at 28°C with a 16h photoperiod. Different growth / incubation durations were studied: 10/3; 13/7; 21/14 and 39/32. For MALDI imaging experiments, the ITO slide was removed from the agar and dried in a dessiccator under vacuum. (HCCA, 5mg/mL in ACN/0.2% TFA 70:30) was used as matrix. UltraFlex II TOF/TOF and Solarix FT-ICR mass spectrometers were used to record molecular cartographies and perform MS/MS experiments for structural analysis purposes. The average mass spectra recorded around the tomato root (2-3 mm on both sides of the root) showed that lipopeptides were major compounds detected on the agar. The relative intensity of lipopeptides families varied with respect to the age of the root/biofilm system. In the 10/3 system, 3 homologues of surfactins were essentially detected (C13, C14 and C15), with very few iturins and fengycins. Their localizations were identical, whatever the considered homologue. Then the production of iturin and fengycin families increases in older systems (13/7 and 21/14) and a novel homologue of surfactin is detected (C12). Some variations in localizations within families may be observed (around the root or at the close vicinity of it in function of the considered homologue or alkali adduct). Then for the oldest system we studied, iturins and fengycins are not detected anymore and the localization of surfactins is less precise. In the 39/32 system, we also detected unknown compounds at 986.6, 1000.6, 1014.7 and 1028.7 m/z. The mass range of these compounds allied to the mass difference between two consecutive ion peaks let us think that these unknown compounds could be a new lipopeptide family. Tandem mass spectrometry experiments, performed on the dried culture medium, allowed to partially sequence these new lipopeptides. MS/MS results allied to exact mass measurements and isotopic pattern simulation give good confidence in the chemical structure we suggest. Nevertheless, to fully identify these new variants of surfactin, micro-extractions followed by (LC)-nano-ESI-MS/MS using a LESA module are in progress. MALDI Mass Spectrometry Imaging becomes a tool to decipher inter-species molecular communication. [less ▲]

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See detailDe novo sequencing of unusual non tryptic peptides thanks to 4-sulfophenylisothiocyanate derivatization by post-source decay MALDI-MS.
Echterbille, Julien ULg; Quinton, Loïc ULg; Escoubas, Pierre et al

Poster (2013, June 11)

Introduction Due to the specificity of trypsin, tryptic peptides contain basic residues on the C-terminal side. This feature provides good ionization efficiency, and facilitates fragmentation processes ... [more ▼]

Introduction Due to the specificity of trypsin, tryptic peptides contain basic residues on the C-terminal side. This feature provides good ionization efficiency, and facilitates fragmentation processes. In the case of non tryptic peptides, the absence of basic residues at one extremity implicates lower fragmentation ratio and poor MS/MS spectra. Several methods have been developed to circumvent this drawback. Derivatization of peptides with compounds containing positive charge has been studied; Chen et al. (RCMS, 2004, 18, 191) demonstrated the simplification of CID spectra of tryptic peptides modified by 4-sulfophenylisothiocyanate. The result is a predominance of y-type ions. In this work, we evaluate the potential of SPITC for the de novo sequencing of unknown non-tryptic peptides containing disulfide bridges, i.e. peptide toxins from animal venoms. Methods 2µL of peptide solution (100 µM) were diluted in 6µL NH4HCO3 50mM (pH 8.7). As peptide toxins often contain disulfide bridges, reduction (2µL DTT 50mM, 1h at 56°C) and alkylation (2µL IAA 500mM, 1h in darkness at RT) of peptides were performed before the derivatization reaction. Peptides were then adsorbed on a C18 ZipTip micro-column followed by 10 µL of 4-sulfophenylisothiocyanate (SPITC) 50mM. The column was then incubated for 6h at 56°C. Peptides were washed by TFA 0.2% and eluted in 10µL 50/50 ACN/FA 0.1%, before being spotted in 2,5-DHB. MS experiments were performed using a Bruker Ultraflex II MALDI-TOF/TOF. FlexControl 3.0, FlexAnalysis 3.0, BioTools 3.2 and SequenceEditor 3.2 softwares (Bruker Daltonics, Bremen) were used for data acquisition and interpretation. Preliminary data According to our first results, SPITC derivatization allows in positive mode to direct the fragmentation thanks to the acidic character of the sulfonate moiety present on the modified molecule. Indeed, a large series of y-type ions is found in the CID spectra allowing determining easily large sequence tags. Moreover, the number of C-terminus ions (b- and a-type ions) decreases, which improve the simplification of MS/MS spectra. Due to this fragmentation pattern, SPITC derivatization is clearly valuable for the sequencing of peptides that are not described in databases (de novo sequencing). For example, animal venoms are composed of several hundreds of peptides that are poorly studied, up to now. These peptides display a high importance for pharmaceutical applications and their sequencing is, as a consequence, of prime interest. Peptide toxins, which are not resulting from an enzymatic digestion, are however difficult to sequence by classical MS/MS methods. In this work, we demonstrate that the modification of peptide toxins with SPITC reagent is suitable for “real” de novo sequencing. The method was applied to isolated peptides as well as chromatographic fractions that contain up to 30 toxins. The perspectives of this work rest on the study of the SPITC modified peptides in negative mode. We expect to obtain a better sensitivity due to the presence of the negative sulfonic acid group at the N-terminus extremity, and also interesting MS/MS spectra including mainly a- or b-type ions. The final challenge will be the application of the protocol to high throughput sequencing of peptide toxins from a large variety of animal venoms. Novel aspect De novo sequencing of unusual non-tryptic peptides thanks to 4-sulfophenylisothiocyanate derivatization by post-source decay MALDI-MS [less ▲]

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See detailIsolation and cultivation of cellulolytic and xylanolytic bacteria and molds extracted from the gut of the termite Reticulitermes santonensis (3DV.1.14)
Tarayre, Cédric ULg; Bauwens, Julien ULg; Mattéotti, Christel et al

Poster (2013, June)

Biofuel production can be based on the use of agro-residues, consisting in a complex lignocellulosic structure which is not easily hydrolysable. The digestive tract of the termite Reticulitermes ... [more ▼]

Biofuel production can be based on the use of agro-residues, consisting in a complex lignocellulosic structure which is not easily hydrolysable. The digestive tract of the termite Reticulitermes santonensis contains a diversified microflora able to hydrolyze the wood components. Bacteria, molds and protists form efficient consortia, able to break the lignocellulosic complex by producing enzymes, such as xylanases and cellulases. Our purpose is the isolation of microbial strains from termite guts in order to evaluate their potential for hydrolysis of lignocellulosic materials. Termites were fed using different diets chosen to improve the xylanolytic and cellulolytic microflora: wood, microcristalline cellulose (added with lignin or not), α-cellulose (added with lignin or not) and birchwood xylan. Then, dissections were realized to isolate the potential xylanolytic and cellulolytic strains. This approach led us to isolate and to study several strains of bacteria (Bacillus sp. strain CTGx and Chryseobacterium sp. strain CTGx) and molds (Trichoderma virens strain CTGx and Sarocladium kiliense strain CTGx). These microorganisms were able to hydrolyze starch, xylan, cellulose, carboxymethylcellulose, esculin, β-glucan and Whatman® filter paper. They can produce glucose and xylose monomers and oligomers which can be further fermented to produce bioethanol. [less ▲]

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See detailResearch of New Enzyme Producing Strains in the Gut of the Termite Reticulitermes santonensis
Tarayre, Cédric ULg; Bauwens, Julien ULg; Mattéotti, Christel et al

Poster (2013, June)

Termites contain a complex microflora inside of their guts. Inferior termites contain bacteria, mycetes and protists that interact to degrade vegetable components. These strains act as consortia to break ... [more ▼]

Termites contain a complex microflora inside of their guts. Inferior termites contain bacteria, mycetes and protists that interact to degrade vegetable components. These strains act as consortia to break natural materials by secreting various enzymes. Our aim was the isolation and cultivation of microorganisms in order to produce new enzymes that can be further used in green chemistry. Termites were fed with different diets: pinewood, microcristalline cellulose (added with lignin or not), α-cellulose (added with lignin or not) and birchwood xylan. Then, dissections were realized to isolate interesting strains. All the microorganisms were subjected to enzyme assays. That technique allowed us to isolate and to cultivate various strains of bacteria, molds and protists. Three strains of bacteria, two strains of molds and one strain of protist were isolated and displayed different enzymatic activities. The bacteria Bacillus subtilis strain ABGx, Bacillus sp. strain CTGx and Chryseobacterium sp. strain CTGx displayed amylase, cellulase and xylanase activities. The molds Trichoderma virens strain CTGx and Sarocladium kiliense strain CTGx were also able to produce those enzymes. However, the protist Poterioochromonas sp. was found to produce only amylase. In conlusion, the termite gut is a complex culivation medium that provides a habitat for many microorganisms that show interesting enzymatic activities. [less ▲]

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See detailDevelopment of a quantitative approach to measure phospholipids in dried drops by Raman spectroscopy
Malherbe, Cédric ULg; Jadoul, Laure ULg; Gilbert, Bernard ULg et al

Poster (2013, May 24)

Phospholipids, PL, such as the phosphatidylcholine PC(18:0/18:1), play a role in the structure of living cells and are suspected to be part of the development of some diseases, for example cancers. Mass ... [more ▼]

Phospholipids, PL, such as the phosphatidylcholine PC(18:0/18:1), play a role in the structure of living cells and are suspected to be part of the development of some diseases, for example cancers. Mass spectrometry enables the structural analysis of PL in complex biological media but imaging mass spectrometry by MALDI-MS is rather limited for quantification purposes. Complementarily, Raman spectroscopy as a non invasive and non destructive method is a potential candidate to quantify and visualise the spatial distribution of the PL by molecular imaging. Unfortunately, the lack of specific chemical function in PL, compared to others biomolecules, limits the use of Raman spectroscopy in the identification process of those PL in complex biological samples. The results presented here belong to a first study of the application of the Raman analyses on dried residues of PL and mice brain tissue performed in the lab. [less ▲]

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See detailLes venins d'animaux, nouvelle panacée?
Echterbille, Julien ULg; Quinton, Loïc ULg; De Pauw, Edwin ULg

in Athena (2013)

Araignées, serpents, scorpions,… autant d’animaux ayant une place particulière dans l’imaginaire collectif. Fascinants, horripilants voire même terrifiants, les adjectifs ne manquent pas pour qualifier ... [more ▼]

Araignées, serpents, scorpions,… autant d’animaux ayant une place particulière dans l’imaginaire collectif. Fascinants, horripilants voire même terrifiants, les adjectifs ne manquent pas pour qualifier les réactions qu’ils suscitent auprès des populations. Que dire alors de la peur engendrée par leur venin. Une simple piqûre, morsure ou contact peut s’avérer extrêmement dangereux voire létal... [less ▲]

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See detailIntraocular lenses with functionalized surfaces by biomolecules in relation with lens epithelial cell adhesion
Huang, Yi-Shiang ULg; Alexandre, Michaël ULg; Bozukova, Dimitriya et al

Poster (2013, April 25)

A cataract is pathology opacity of the lens or capsule of the eye, causing impairment of vision or even blindness. Surgery, with lens extraction and intraocular lens implantation, is still the only ... [more ▼]

A cataract is pathology opacity of the lens or capsule of the eye, causing impairment of vision or even blindness. Surgery, with lens extraction and intraocular lens implantation, is still the only currently available treatment. The most common complication after implantation of intraocular lenses (IOLs) is the posterior capsular opacification (PCO) or secondary cataract. This is the result of lens epithelial cells (LECs) proliferation and their transition to mesenchymal cells. In 1997, a Sandwich theory was proposed to elucidate the developmental process of PCO. [1] According to this model, an IOL with higher affinity to LECs will induce a less PCO. In our research, the pHEMA (Poly(2-hydroxyethyl methacrylate)) based acrylic hydrophilic polymer is subjected to the surface modification by conjugating with the bioactive peptides. The RGD sequence, known for its excellent biocompatibility, is designed to stimulate the biointegration between the LECs and the polymer implant. [2]. From our research, The RGD peptide immobilized onto pHEMA surfaces significantly facilitates the adhesion of the porcine LEC. The peptide immobilized surface retains its biological function even after 10 times of autoclave. On the other hand, the immobilized peptide does not alter the hydrophobicity of the surface, the light transmission, as well as the cytotoxicity of the material. This functionalized biomaterial would possibly prevent the formation of PCO. [1] J Cataract Refract Surg. 1997 Dec;23(10):1539-42 [2] Trends Biotechnol. 2008 Jul;26(7):382-92 [less ▲]

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See detailAdvances in proteomics for the FP7 Venomics project
Degueldre, Michel ULg; Quinton, Loïc ULg; De Pauw, Edwin ULg

Scientific conference (2013, April)

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See detailDevelopment of an analytical method to detect metabolites of nitrofurans. Application to the study of furazolidone elimination in Vietnamese black tiger shrimp (Penaeus monodon)
Douny, Caroline ULg; Widart, Joëlle ULg; De Pauw, Edwin ULg et al

in Aquaculture (2013), 376-379

Nitrofurans, banned antibiotics in European Union (EU), have often been found in imported aquaculture products in the past and are still found nowadays according to the Rapid Alert System for Feed and ... [more ▼]

Nitrofurans, banned antibiotics in European Union (EU), have often been found in imported aquaculture products in the past and are still found nowadays according to the Rapid Alert System for Feed and Food (RASFF) of the European Commission. A quantitative method based on liquid chromatography coupled to isotopic dilution tandem mass spectrometry (LC–IDMS/MS) was developed for the determination of the residues of four nitrofuran antibiotic residues in shrimps. The experimental protocol consisted of an acid-catalysed release of protein-bound metabolites, followed by derivatisation with 2-nitrobenzaldehyde (NBA). Then, a double liquid–liquid extraction with ethyl acetate was performed before LC–IDMS/MS analysis by positive electrospray ionisation (ES+) with multiple reaction monitoring (MRM) of two transitions per compound. An “in-house” validation of the method for shrimp analysis was conducted according to the EU criteria for the analysis of veterinary drug residues in foods. The decision limits (CCalpha) were 0.08–0.36 µg kg -1 and the detection capabilities (CCbeta) were 0.12–0.61 µg kg-1, which are both below the minimum required performance limit (MRPL) set at 1 µg kg -1 by the EU. The developed method was applied to evaluate the elimination of furazolidone residues in shrimp muscles after a contamination experiment. After 28 days of decontamination, a concentration of 115 µg kg-1 of furazolidone metabolite 3-amino-2-oxazolidinone (AOZ) was still measured in shrimp muscle. [less ▲]

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See detailDiscrimination of Isobaric Leu/Ile Residues by MALDI In-source Decay Mass Spectrometry
Asakawa, Daiki; Smargiasso, Nicolas ULg; De Pauw, Edwin ULg

in Journal of the American Society for Mass Spectrometry (2013), 24(2), 297-300

MALDI in-source decay (ISD) has been used for the top-down sequencing of proteins. The use of 1,5-diaminonapthalene (1,5-DAN) gave strong intensity of w ions, which are informative fragments and can be ... [more ▼]

MALDI in-source decay (ISD) has been used for the top-down sequencing of proteins. The use of 1,5-diaminonapthalene (1,5-DAN) gave strong intensity of w ions, which are informative fragments and can be helpful for the distinction of the isobaric amino acids, Leu and Ile. Our data suggests that the w fragments are formed from z* radical fragment by unimolecular dissociation and high abundance of w ions in MALDI-ISD with 1,5-DAN can be understood as resulting from the low collision rate in the MALDI plume. The MALDI-ISD with 1,5-DAN could be a useful method for the top-down sequencing of proteins including discrimination of Leu and Ile near the C-terminal end. [less ▲]

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See detailUltraviolet Laser Induced Hydrogen Transfer Reaction: Study of the First Step of MALDI In-Source Decay Mass Spectrometry
Asakawa, Daiki; Calligaris, David ULg; Smargiasso, Nicolas ULg et al

in Journal of Physical Chemistry B (2013), 117(8), 2321-2327

The early mechanisms of matrix-assisted laser desorption/ionization in-source decay (MALDI-ISD) are described herein. MALDI-ISD is initiated by the hydrogen transfer from excited matrix molecules to the ... [more ▼]

The early mechanisms of matrix-assisted laser desorption/ionization in-source decay (MALDI-ISD) are described herein. MALDI-ISD is initiated by the hydrogen transfer from excited matrix molecules to the carbonyl oxygen of the peptide backbone, which is followed by a radical-induced cleavage, producing the c′/z• fragment pair. As expected, the use of 2,5-DHB or 1,5-DAN was efficient to induce MALDI-ISD, and the strongest intensity of MALDI-ISD fragments was observed when laser shots were performed on matrix crystals. In contrast, the hydrogen radical transfer reaction was suppressed by using ionic liquid and amorphous structure of 2,5-DHB and 1,5-DAN mixture as a matrix. Our results suggest that the hydrogen transfer occurs on the matrix crystal during the dissipation of the laser energy and before desorption, following ISD fragments formed in the MALDI plume. [less ▲]

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