Purification, crystallization and preliminary crystallographic analysis of the catalytic domain of the extracellular cellulase CBHI from Trichoderma harzianum.
The filamentous fungus Trichoderma harzianum has a considerable cellulolytic activity that is mediated by a complex of enzymes which are essential for the hydrolysis of microcrystalline cellulose. These enzymes were produced by the induction of T. harzianum with microcrystalline cellulose (Avicel) under submerged fermentation in a bioreactor. The catalytic core domain (CCD) of cellobiohydrolase I (CBHI) was purified from the extracellular extracts and submitted to robotic crystallization. Diffraction-quality CBHI CCD crystals were grown and an X-ray diffraction data set was collected under cryogenic conditions using a synchrotron-radiation source.
Colussi
Francieli
F
Instituto de Física de São Carlos, Universidade de São Paulo, Avenida Trabalhador Saocarlense 400, 13560-970 São Carlos-SP, Brazil.
Textor
Larissa C
LC
Serpa
Viviane
V
Maeda
Roberto Nobuyuki
RN
Pereira
Nei
N
Polikarpov
Igor
I
D020134:Catalytic Domain; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000737:chemistry* / Q000302:isolation & purification; D003461:Crystallography; D018360:Crystallography, X-Ray; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D014242:Trichoderma / Q000201:enzymology*
D005656:Fungal Proteins; D043366:Cellulose 1,4-beta-Cellobiosidase
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