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In this work, a new strategy of biosensor design is developed based on the assembly of amyloid beta and its multiple interactions with other bioactive species. These interactions can enable amyloid beta peptide as a multifunctional sensing element, so the immobilization of sensing probe and the step-by-step modification of the sensing interface have all been dispensed with. Instead, the kinetics of the assembly of a peptide-based catalytic network serves to convert the quantity of analyte into amplified signal readout. The designed dynamic assembling and biosensing system has also been successfully applied in detecting the activity of polyglutamylation, an essential post translation modification controlling cell skeleton and cell cycle, in biological complex samples. Further studies reveal that the serum abundance of a polyglutamylase, tubulin tyrosine ligase-like protein 12, may show parallel with the degree of development of prostate cancer and the discrimination between early cancerous development and benign conditions. And the obtained result is more distinct than that based on PSA detection, the current gold standard. This study may also point to the prospective of extending this design strategy to broader range of biosensing applications in the future.

Allozyme variation was assessed in a collection of pearl millet cultivars derived from three different regions of traditional cultivation of this cereal in Tunisia and West African countries. Using starch gel electrophoresis, 11 loci corresponding to seven enzyme staining systems were identified. Important within-cultivar genetic diversity was detected. Genetic diversity within Tunisian cultivars was similar and was more pronounced than for West African ones. This result could be explained by the low level of the human selection characterizing the Tunisian cultivars. Most of the total genetic diversity is maintained within subregions suggesting little genetic differentiation. Cluster analysis did not reveal separate clusters for the various subregion cultivars. However the cluster analysis should help in the selection of interesting genotypes for future breeding programs.

Peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidase F (PNGase F) and endo-β-N-acetyl glucosaminidase F (Endo F) activities were monitored during cultivation ofFlavobacterium meningosepticumusing a new fluorescence-HPLC procedure based on a commercially available substrate. The PNGase F activity reached a maximum level at the end of the log phase and remained constant during the stationary phase, while Endo F continuously increased until late stationary phase. PNGase F obtained at the end of the log phase was less contaminated by other proteins compared with late stationary phase.

We have previously isolated mannoside and xylomannoside oligosaccharides with one or two terminal reducingN-acetylglucosamine residues from the extracellular medium of white campion (Silene alba) suspension culture. We have now demonstrated the presence of peptide-N4-(N-acetylglucosaminyl)asparagine amidase (PNGase) activity in cell extracts as well in the culture medium that could explain the production of those compounds. An additional xylomannoside, (GlcNAc)Man3(Xyl)GlcNAc(Fuc)GlcNAc, was characterized, and1H- and13C-NMR assignments for the oligosaccharide Man3(Xyl)GlcNAc(Fuc)GlcNAc were obtained using homonuclear and heteronuclear spectroscopy (COSY).

K99 lectin fromEscherichia coliwas purified and biotinylated via its carboxyl groups using biocytin hydrazide and a water soluble carbodiimide. Biotinylation of two out of the nine carboxyl groups was sufficient to permit detection of the lectin by avidin and did not cause any loss of the haemagglutinating activity. It was demonstrated that the biotinylated K99 lectin retained other important properties of native K99 and that it will probably become a very sensitive detecting reagent. Indeed, it was able to bind to HeLa cells, as do intact bacteria carrying K99 fimbriae, and also to recognizeN-glycolyl-neuraminyl-lactosyl-ceramide in an overlay binding assay.

The peptide-N4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) earlier described [Lhernould S., Karamanos Y., Bourgerie S., Strecker G., Julien R., Morvan H. (1992)Glycoconjugate J9:191–97] was partially purified from culturedSilene albacells using affinity chromatography. The enzyme is active between pH 3.0 and 6.5, and is stable in the presence of moderate concentrations of several other protein unfolding chemicals, but is readily inactivated by SDS. Although the enzyme cleaves the carbohydrate from a variety of animal and plant glycopeptides, it does not hydrolyse the carbohydrate from most of the corresponding unfolded glycoproteins in otherwise comparable conditions. The substrate specificity of this plant PNGase supports the hypothesis that this enzyme could be at the origin of the production of ‘unconjugated N-glycans’ in a suspension medium of culturedSilene albacells.

In this study it was shown that the O101 lipopolysaccharide isolated from Escherichia coli B41 did not contain an O-specific polysaccharide and that its sugar moiety is probably restricted to the core oligosaccharide. It is characterized by the presence of galactose, glucose, N-acetylglucosamine, heptose and 3-deoxy-d-manno-2-octulosonic acid and the fatty acid composition is typical of an Enterobacteriaceae lipopolysaccharide. Methylation analysis indicated terminal non-reducing galactose and glucose and also 1,2-linked glucose which is a substitution pattern typical of an E. coli lipopolysaccharide core oligosaccharide. The obtained structural information is sufficient to explain the previously observed interactions between the O101 lipopolysaccharide and the K99 lectin.