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Visualization of cellulases bound to cellulose microfibrils: Evidence for endo/exo synergism

Conference ·
OSTI ID:141961
;  [1];  [2]
  1. NREL, Golden, CO (United States)
  2. Colorado State Univ., Fort Collins, CO (United States)
Monoclonal antibodies (MAbs) specific for cellobiohydrolase I (CBH I) and endoglucanase I (EG I) were conjugated to 10-nm and 15-nm colloidal gold particles, respectively. The binding of CBH I and EG I to cellulose was visualized by utilizing the MAb-colloidal gold probes. The visualization procedure involved immobilization of cellulose microfibrils on copper EM grids, incubation of the cellulose-coated grids with cellulase(s), binding of MAb-colloidal gold conjugates to cellulase(s), and visualization via transmission electron microscopy (TEM). Apparent differences in the substrate binding preferences for gold-labelled CBH I and EG I were observed; however, the actual structure of the cellulose at these sites was not confirmed. Sites showing binding of both enzymes sometimes showed patterns of coordination suggestive of the {open_quotes}progressive{close_quotes} cellulase action model. The sensitivity of this technique was evident when micrographs were observed in 3-D.
OSTI ID:
141961
Report Number(s):
CONF-930304--
Country of Publication:
United States
Language:
English

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