Cell Grow (21)

Growth of C. thermocellum on cellobiose was measured as a function of optical density by

spectrophotometry (Novaspec II; Biochrom Ltd., Cambridge, UK) at 600 nm (OD600)
immediately after briefly vortexing the tube. A dry mass measurement of approximately 0.5 g/L
was found to be correlated with an OD600 of 1, which is in agreement with previous observations
(Payot et al. 1998). Cell growth on insoluble -cellulose was determined indirectly by measuring
total protein content of samples, using a modification of the
Bradford method (Bradford 1976). Samples (10 mL) were centrifuged (8000g for 15 min) and
supernatant was removed. Pellets were washed with 0.9% (m/v) NaCl and resuspended in 2 mL
of 0.2 mol/L NaOH. Suspensions were incubated 10 min in a boiling water bath. After cooling
the samples were centrifuged (8000g for 15 min) and the supernatants were collected for
protein assays (Desvaux et al. 2001).