Vignette d'image

Neuhaus, Jean-Marc

Nom
Neuhaus, Jean-Marc
Affiliation principale
Fonction
Professeur ordinaire
Email
jean-marc.neuhaus@unine.ch
Identifiants
https://libra.unine.ch/handle/123456789/300

Résultat de la recherche

Filtres

2
2
2
Réinitialiser les filtres

Paramètres

Voici les éléments 1 - 2 sur 2
  • Publication
    Métadonnées seulement
    Identification of novel secreted proteases during extracellular proteolysis by dermatophytes at acidic pH
    (2011)
    Sriranganadane, D.
    ;
    Waridel, P.
    ;
    Salamin, K.
    ;
    Feuermann, M.
    ;
    Mignon, B.
    ;
    Staib, P.
    ;
    Neuhaus, Jean-Marc 
    ;
    Quadroni, M.
    ;
    Monod, M.
    The dermatophytes are a group of closely related fungi which are responsible for the great majority of superficial mycoses in humans and animals. Among various potential virulence factors, their secreted proteolytic activity attracts a lot of attention. Most dermatophyte-secreted proteases which have so far been isolated in vitro are neutral or alkaline enzymes. However, inspection of the recently decoded dermatophyte genomes revealed many other hypothetical secreted proteases, in particular acidic proteases similar to those characterized in Aspergillus spp. The validation of such genome predictions instigated the present study on two dermatophyte species, Microsporum canis and Arthroderma benhamiae. Both fungi were found to grow well in a protein medium at acidic pH, accompanied by extracellular proteolysis. Shotgun MS analysis of secreted protein revealed fundamentally different protease profiles during fungal growth in acidic versus neutral pH conditions. Most notably, novel dermatophyte-secreted proteases were identified at acidic pH such as pepsins, sedolisins and acidic carboxypeptidases. Therefore, our results not only support genome predictions, but demonstrate for the first time the secretion of acidic proteases by dermatophytes. Our findings also suggest the existence of different pathways of protein degradation into amino acids and short peptides in these highly specialized pathogenic fungi.
  • Publication
    Métadonnées seulement
    High-yield production and purification of recombinant T7-tag mature streptavidin in glucose-stressed E. coli
    (2008)
    Humbert, N.
    ;
    Schurmann, P.
    ;
    Zocchi, A.
    ;
    Neuhaus, Jean-Marc 
    ;
    Ward, T. R.
    The overexpression of toxic recombinant proteins is often problematic, leading to either low production levels or inclusion bodies. Streptavidin is no exception and thus the highest production level reported to date for streptavidin is 70 mg/L of functional protein. Herein, we report on the production in Escherichia coli and the purification of a recombinant mature streptavidin bearing a T7-tag. Optimization of critical parameters, including the glucose concentration, the pH and the time of induction as well as the use of BL21(DE3)pLysS cell strain, affords up to 120 mg/L functional streptavidin in soluble form. The yield can be further increased by an osmotic stress during the preculture by adding highly concentrated glucose before the inoculation of the culture medium, thus affording reproducibly 230 mg/L of soluble streptavidin. A single denaturing-renaturing step and affinity chromatography afford highly active tetrameric protein with >3.8/4.0 active sites.