The cellulosomes from Clostridium cellulolyticum: identification of new components and synergies between complexes.
Cellulosomes produced by Clostridium cellulolyticum grown on cellulose were purified and separated using anion-exchange chromatography. SDS/PAGE analysis of six fractions showed variations in their cellulosomal protein composition. Hydrolytic activity on carboxymethyl cellulose, xylan, crystalline cellulose and hatched straw differed from one fraction to another. Fraction F1 showed a high level of activity on xylan, whereas fractions F5 and F6 were most active on crystalline cellulose and carboxymethyl cellulose, respectively. Several cellulosomal components specific to fractions F1, F5 and F6 were investigated using MS analysis. Several hemicellulases were identified, including three xylanases in F1, and several cellulases belonging to glycoside hydrolase families 9 and 5 and, a cystein protease inhibitor were identified in F5 and F6. Synergies were observed when two or three fractions were combined. A mixture containing fractions F1, F3 and F6 showed the most divergent cellulosomal composition, the most synergistic effects and the highest level of activity on straw (the most heterogeneous substrate tested). These findings show that on complex substrates such as straw, synergies occur between differently composed cellulosomes and the degradation efficiency of the cellulosomes is correlated with their enzyme diversity.
Fendri
Imen
I
Laboratoire de Chimie Bactérienne, CNRS, UPR9043, IMM, Marseille, France.
Tardif
Chantal
C
Fierobe
Henri-Pierre
HP
Lignon
Sabrina
S
Valette
Odile
O
Pagès
Sandrine
S
Perret
Stéphanie
S
D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D002480:Cellulase / Q000302:isolation & purification / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid / Q000379:methods; D046973:Ruminiclostridium cellulolyticum / Q000187:drug effects / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000302:isolation & purification / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000302:isolation & purification / Q000378:metabolism; D002479:Inclusion Bodies; D013058:Mass Spectrometry / Q000379:methods
D001426:Bacterial Proteins; D015853:Cysteine Proteinase Inhibitors; D002482:Cellulose; D006026:Glycoside Hydrolases; C023305:hemicellulase; D002480:Cellulase