For the analysis of solid-state 13C nuclear magnetic resonance (NMR)
and Fourier transform infrared (FTIR) spectroscopy, specimens were prepared
according to the following method. Using forceps, we removed
sheets of cellulose film from cellulose pellicles produced by the control
cells. In the mutant F2-2 culture, particulate material was collected by
centrifugation. The pellet was resuspended in 10 mM K-acetate buffer
(pH 5.0) and recentrifuged at 5,000 rpm for 20 min. In the F2-2 culture,
cells grew rapidly to a large concentration. In order to avoid contamination
of the cell fragments, we washed the resultant pellet with a sodium
hydrate solution. The cellulose product (pellicle or particulate material of
the mutant) was stored in 0.1 M NaOH at 80°C overnight and then
washed several times with hot water. The purified cellulose and alkaliinsoluble
materials were dried at 60°C overnight. Avicel (microcrystalline
cellulose) (Merck, Darmstadt, Germany) was treated with 4 M NaOH at
80°C and washed several times with hot water. These products were analyzed
by solid-state 13C NMR and FTIR spectrometry.