Table II.
Influence of proteinase K on size and composition of (1→3),(1→4)-β-glucan synthesized in vitro
The high- and low-mass fractions pooled for these analyses are shown in Figure 5 (chromatographic profiles of synthase reactions without CHAPS were similar; data not shown). After dialysis against deionized water, the polymers and oligomers were freeze dried, digested with B. subtilis endoglucanase, and cellobiosyl- and cellotetraosyl-(1→3)-glucose oligomers were separated by HPAEC. Radioactivity recovered in each oligomer was corrected for no. of glucose equivalents in the calculation of trimer:tetramer ratios.
| Oligomer
|
No Proteinase
|
0.1 mg mL-1
|
0.2 mg mL-1
|
|||
|---|---|---|---|---|---|---|
| High mass | Low mass | High mass | Low mass | High mass | Low mass | |
| Experiment I: no detergent | ||||||
| Trimer: tetramer | 3.37 | 0.48 | 3.44 | 0.48 | 3.13 | 0.22 |
| % Product | 66.7 | 32.3 | 81.1 | 18.9 | 30.3 | 69.7 |
| Experiment II: + 0.1% (w/v) CHAPS | ||||||
| Trimer: tetramer | 2.75 | 0.44 | 2.44 | 0.14 | 1.88 | 0.21 |
| % Product | 78.3 | 21.7 | 64.1 | 35.9 | 47.9 | 52.1 |