Hydrolysis of amorphous and crystalline cellulose by heterologously produced cellulases of Melanocarpus albomyces.
Three thermostable neutral cellulases from Melanocarpus albomyces, a 20-kDa endoglucanase (Cel45A), a 50-kDa endoglucanase (Cel7A), and a 50-kDa cellobiohydrolase (Cel7B) heterologously produced in a recombinant Trichoderma reesei were purified and studied in hydrolysis (50 degrees C, pH 6.0) of crystalline and amorphous cellulose. To improve their efficiency, M. albomyces cellulases naturally harboring no cellulose-binding module (CBM) were genetically modified to carry the CBM of T. reesei CBHI/Cel7A, and were studied under similar experimental conditions. Hydrolysis performance and product profiles were used to evaluate hydrolytic features of the investigated enzymes. Each cellulase proved to be active against the tested substrates; the cellobiohydrolase Cel7B had greater activity than the endoglucanases Cel45A and Cel7A against crystalline cellulose, whereas in the case of amorphous substrate the order was reversed. Evidence of synergism was observed when mixtures of the novel enzymes were applied in a constant total protein dosage. Presence of the CBM improved the hydrolytic potential of each enzyme in all experimental configurations; it had a greater effect on the endoglucanases Cel45A and Cel7A than the cellobiohydrolase Cel7B, especially against crystalline substrate. The novel cellobiohydrolase performed comparably to the major cellobiohydrolase of T. reesei (CBHI/Cel7A) under the applied experimental conditions.
Szijártó
Nóra
N
Department of Applied Biotechnology and Food Science, Budapest University of Technology and Economics, P.O. Box 91, H-1521 Budapest, Hungary. nora_szijarto@mkt.bme.hu
Siika-Aho
Matti
M
Tenkanen
Maija
M
Alapuranen
Marika
M
Vehmaanperä
Jari
J
Réczey
Kati
K
Viikari
Liisa
L
D001203:Ascomycota / Q000201:enzymology*; D002480:Cellulase / Q000737:chemistry* / Q000302:isolation & purification; D002482:Cellulose / Q000737:chemistry*; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D006868:Hydrolysis; D007700:Kinetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
D005656:Fungal Proteins; D011994:Recombinant Proteins; D002482:Cellulose; D002480:Cellulase