Refinement of the production of antigen-specific hen egg yolk antibodies (IgY) intended for passive dietary immunization in animals: A review

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (in press)

Antibodies have become essential tools in recent decades, with a wide range of applications in the laboratory and in human and veterinary medicine. The use of laying hens, instead of mammals, to obtain ... [more ▼]

Antibodies have become essential tools in recent decades, with a wide range of applications in the laboratory and in human and veterinary medicine. The use of laying hens, instead of mammals, to obtain the necessary antibodies from the eggs is a major advance in terms of animal welfare because it makes blood sampling obsolete. However, the advantages of this technology are numerous, in addition to the animal welfare aspect. With a carefully designed immunization protocol, it is possible to enhance both the hen’s immune response and its welfare during the process. This review puts forward recommendations how to do this and discusses recent approaches on improving the technology. [less ▲]

Characterization of new bacterial glycoside hydrolases isolated from agricultural soils using a functional metagenomic approach

Poster (2013, June 10)

Microorganisms play key roles in soil ecosystem functioning, notably through their ability to degrade plant cell wall polymers. For this, bacteria and fungi produce various enzymes such as cellulases ... [more ▼]

Microorganisms play key roles in soil ecosystem functioning, notably through their ability to degrade plant cell wall polymers. For this, bacteria and fungi produce various enzymes such as cellulases, xylanases, glucosidases, esterases or laccases. Finding new enzymes hydrolyzing cellulose, hemicellulose or lignin is not only interesting for a better understanding of the roles of the soil microflora still largely unknown but these enzymes are also useful for various biotechnological applications such as the production of renewable energy from lignocellulosic material. So here, we used a functional metagenomic approach to isolate new bacterial β-glucosidases, which were then biochemically characterized. The new enzymes were identified by functional analysis of agricultural-soil metagenomic libraries hosted in Escherichia coli and screened on medium containing esculin. After sequence analysis and preliminary estimation of the activity of the new β-glucosidases using p-nitrophenol derivatives on intact bacterial cells, the coding sequences of three of them were cloned into a bacterial expression vector so as to overproduce and purify them by affinity chromatography. The chosen enzymes show only 52-64% sequence identity to known family 3 (GH3) or 1 (GH1) glycoside hydrolases of different phyla (Actinobacteria, Acidobacteria and Proteobacteria). Analysis of the E. coli cells expressing each of them revealed that both GH1 proteins (ASEsc9 and ASEsc10) are thermophilic enzymes more active at mildly acidic to neutral pH while the GH3 enzyme (ASEsc6) is an alkaline, mesophilic, β-glucosidase also displaying xylosidase activity. Their coding sequences have been cloned in fusion with a carboxy-terminal His-tag and placed under the control of the IPTG-inducible promoter of the pET-30b vector. The proteins will be overproduced and purified for further characterization. [less ▲]

Biomass hydrolyzing enzymes identified by functional screening of a metagenomic library from algal biofilms.

Poster (2013, June)

Biomass hydrolyzing enzymes are increasingly searched for the production of biofuels and renewable chemical compounds using biomass. Microorganisms living on algaes are an interesting reservoir of biomass ... [more ▼]

Biomass hydrolyzing enzymes are increasingly searched for the production of biofuels and renewable chemical compounds using biomass. Microorganisms living on algaes are an interesting reservoir of biomass hydrolyzing enzymes, as they are in constant interaction with algal biomass. Therefore, we are interested in looking for some of those enzymes synthesized by the microflora living on the surface of the brown algae Ascophyllum nodosum. Algae samples were collected in the winter 2012 and a microbial DNA extraction method was developed. The whole extracted microbial genomes of the microorganisms living on the algae were restricted, inserted in a cloning vector and ligated products were used for transformation of cultivable Escherichia coli host cells. This metagenomic library was then screened for diverse enzymatic activities (lipolytic enzymes, cellulases, beta-glucosidases, alpha-amylases, arabinanases, xylanases and proteases) on agar plates with specific substrates. Five putative lipolytic enzymes, one cellulase and one beta-glucosidase were identified. Sequence analysis revealed low (<50%) sequence identities with known enzymes sequences, meaning new enzymes from unknown genomes have been discovered. To our knowledge this is the first functional screening that was realized with a metagenomic library from algal biofilms and this is the first cellulase identified by marine metagenomics. A second library has been constructed from algae sample from summer 2012 and is currently being screened. New enzymatic tests are being developed for the identification of enzymes degrading specific algal polysaccharides like agarases, carrageenases, alginate lyases, laminarinases,… Those very specific enzymes aren’t well known yet, and our metagenomic approach will probably help us to identify new families and structures of those algal biomass hydrolyzing enzymes. [less ▲]

D31-1238: Développement et utilisation des anticorps du jaune d'oeuf en vue du contrôle des Salmonella spp. dans le tube digestif du poulet de chair (Rapport final)

Report (2013)

Functional screening of a metagenomic library from algal biofilms

in Smagghe, Guy; Boeckx, Pascal; Bossier, Peter (Eds.) et al Communications in Agricultural and Applied Biological Sciences (2013, February 08)

Macroalgae, and particularly their lignin-free polysaccharides, are increasingly used for their gelling and therapeutic properties and for the production of biofuels and renewable chemical compounds. To ... [more ▼]

Macroalgae, and particularly their lignin-free polysaccharides, are increasingly used for their gelling and therapeutic properties and for the production of biofuels and renewable chemical compounds. To extract, hydrolyze and purify this biomass, algae hydrolyzing enzymes are needed. Our work aims to identify and characterize algal biomass hydrolyzing enzymes expressed by microorganisms living on the surface of algae, by functional metagenomics. Therefore, a microbial DNA extraction method was developed to isolate the gDNA from the microorganisms of the brown algae Ascophyllum nodosum and a metagenomic library was constructed in Escherichia coli. The library was screened for diverse enzymatic activities (esterases, xylanases, cellulases, α-amylases, arabinanases, caseinases and β-glucosidases) on agar plates with specific enzymes substrates. Several new microbial enzymes (esterases, β-glucosidases, α-amylases and cellulases) were identified revealing the wealth of our library. Furthermore, those enzymes had less than 50% sequence identity with known protein sequences; meaning that our approach allows to identify new microbial enzymes expressed by uncultured microorganisms. Plate tests for medium-throughput screening of specific enzymes hydrolyzing algal polysaccharides (agarases, carrageenases and alginate lyases) are currently being developed. Our approach will probably allow us to identify new families of those ill-known enzymes, with particular enzymatic activities. [less ▲]

An eYFP Reporter Gene for the Yeast Two-hybrid System

in Protein Journal (2013), 32(2), 126-130

Influence of lignin in Reticulitermes santonensis: symbiotic interations investigated through proteomics

in Symbiosis (2013)

The gut of lower termites is populated by numerous microbial species belonging to prokaryotes, fungi, yeasts and protists. These micro-organisms are organized in a complex symbiotic system, interacting ... [more ▼]

The gut of lower termites is populated by numerous microbial species belonging to prokaryotes, fungi, yeasts and protists. These micro-organisms are organized in a complex symbiotic system, interacting together and with the insect host. Their likely ability to degrade ligno-cellulosic compounds could lead to improvements in second generation biofuels production. Lignin elimination represents a critical point as this polymer significantly interferes with industrial process of cellulose. Although host produces its own lignin-degrading enzymes, some symbionts may participate in digestion of lignin and its degradation products in termite gut. Here, we compared gut proteomes from R. santonensis after rearing on artificial diets composed of cellulose with and without lignin. The effect of lignin in artificial diets on different parts of the digestive tract was compared through liquid chromatography associated with tandem mass spectrometry (LC-MS/MS) experiments. Enzymatic assays were performed to characterize activities present in R. santonensis digestive tract after feeding on artificial diets. Microscopic observations of microbial communities provided some information on population balances after feeding experiment. [less ▲]

Discrepancies in microbiota composition along the pig gastrointestinal tract between in vivo observations and an in vitro batch fermentation model

in Journal of Animal Science (2012), 90(supplement 4), 393-396

Termites as a tool to improve lignocellulose biomass valorization : study of enzymatic complex in termites and its common symbionts by comprehensive metabolite profiling

Conference (2012, November 16)

Improved in vitro gas fermentation method to assess prebiotic potential of indigestible carbohydrates

Poster (2012, November 09)