Characterisation of endoglucanases EGB and EGC from Fibrobacter succinogenes - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Accéder directement au contenu
Article Dans Une Revue Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology Année : 2000

Characterisation of endoglucanases EGB and EGC from Fibrobacter succinogenes

Résumé

The enzymatic properties of two endoglucanases from Fibrobacter succinogenes, EGB and EGG, were analysed. EGB and EGC were purified from recombinant Escherichia coli cultures expressing their gene. The failure of purification of EGB by classical techniques led us to produce antipeptide antibodies that allowed immunopurification of the protein from E. coli as well as its detection in F. succinogenes cultures. Synthetic peptides were selected from the predicted primary structure of EGB, linked to bovine serum albumin and used as immunogens to obtain specific antibodies. One of the polyclonal antipeptide antisera was used to purify EGB. EGC was purified by affinity chromatography with Ni-NTA resin. The endo mode of action of the two enzymes on carboxymethyl-cellulose was different. The values of K-m and V-max were respectively 13.6 mg/ml and 46 mu mol/min mg protein for EGB, and 7 mg/ml and 110 mu mol/min mg protein for EGG. The reactivity of the antipeptide and the anti-EGG sera with F. succinogenes proteins of molecular mass different from that of EGB and EGC produced in E. coli suggested post-translational modification of the two enzymes in F. succinogenes cultures. Expression of endB and endC genes in F. succinogenes was confirmed by RT-PCR.

Dates et versions

hal-02698440 , version 1 (01-06-2020)

Identifiants

Citer

Christel Béra Maillet, Veronique V. Broussolle, Peter Pristas, Jean-Pierre Girardeau, Geneviève Gaudet, et al.. Characterisation of endoglucanases EGB and EGC from Fibrobacter succinogenes. Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 2000, 1476 (2), pp.191-202. ⟨10.1016/S0167-4838(99)00255-1⟩. ⟨hal-02698440⟩

Collections

INRA INRAE
13 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More