Comparative secretome analyses of two Trichoderma reesei RUT-C30 and CL847 hypersecretory strains.
BACKGROUND
Due to its capacity to produce large amounts of cellulases, Trichoderma reesei is increasingly been researched in various fields of white biotechnology, especially in biofuel production from lignocellulosic biomass. The commercial enzyme mixtures produced at industrial scales are not well characterized, and their proteinaceous components are poorly identified and quantified. The development of proteomic methods has made it possible to comprehensively overview the enzymes involved in lignocellulosic biomass degradation which are secreted under various environmental conditions.


RESULTS
The protein composition of the secretome produced by industrial T. reesei (strain CL847) grown on a medium promoting the production of both cellulases and hemicellulases was explored using two-dimensional electrophoresis and MALDI-TOF or LC-MS/MS protein identification. A total of 22 protein species were identified. As expected, most of them are potentially involved in biomass degradation. The 2D map obtained was then used to compare the secretomes produced by CL847 and another efficient cellulolytic T. reesei strain, Rut-C30, the reference cellulase-overproducing strain using lactose as carbon source and inducer of cellulases.


CONCLUSIONS
This study provides the most complete mapping of the proteins secreted by T. reesei to date. We report on the first use of proteomics to compare secretome composition between two cellulase-overproducing strains Rut-C30 and CL847 grown under similar conditions. Comparison of protein patterns in both strains highlighted many unexpected differences between cellulase cocktails. The results demonstrate that 2D electrophoresis is a promising tool for studying cellulase production profiles, whether for industrial characterization of an entire secretome or for a more fundamental study on cellulase expression at genome-wide scale.
Herpoël-Gimbert
Isabelle
I
INRA, UMR1163, Biotechnologie des Champignons Filamenteux, F-13000 Marseille, France.|Universités Aix-Marseille I & II, UMR1163, BCF, F-13000 Marseille, France.
Margeot
Antoine
A
IFP, Biotechnology Department, Avenue de Bois-Préau, 92852 Rueil-Malmaison Cedex, France.
Dolla
Alain
A
IMR, FRE3083 - CNRS, Institut de Biologie Structurale et Microbiologie, Chemin Joseph-Aiguier, 13402 Marseille cedex 20, France.
Jan
Gwénaël
G
INRA, UMR1253, Science et Technologie du Lait et de l'Oeuf, F-35000 Rennes, France.|Agrocampus Rennes, UMR1253, STLO, F-35000 Rennes, France.
Mollé
Daniel
D
INRA, UMR1253, Science et Technologie du Lait et de l'Oeuf, F-35000 Rennes, France.|Agrocampus Rennes, UMR1253, STLO, F-35000 Rennes, France.
Lignon
Sabrina
S
Plate-forme protéomique, Institut de Biologie structurale et Microbiologie, Chemin Joseph-Aiguier, 13402 Marseille cedex 20, France.
Mathis
Hughes
H
IFP, Biotechnology Department, Avenue de Bois-Préau, 92852 Rueil-Malmaison Cedex, France.
Sigoillot
Jean-Claude
JC
INRA, UMR1163, Biotechnologie des Champignons Filamenteux, F-13000 Marseille, France.|Universités Aix-Marseille I & II, UMR1163, BCF, F-13000 Marseille, France.
Monot
Frédéric
F
IFP, Biotechnology Department, Avenue de Bois-Préau, 92852 Rueil-Malmaison Cedex, France.
Asther
Marcel
M
INRA, UMR1163, Biotechnologie des Champignons Filamenteux, F-13000 Marseille, France.|Universités Aix-Marseille I & II, UMR1163, BCF, F-13000 Marseille, France.