Surface Modification of Cellulose Acetate with Cutinase and ...
Surface Modification of Cellulose Acetate with Cutinase and ...
Surface Modification of Cellulose Acetate with Cutinase and ...
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Strategies Towards the Functionalization <strong>of</strong> Bacillus subtilis Subtilisin E for Wool Finishing Applications<br />
subtilisin was secreted to the periplasmic space, (figures 3 <strong>and</strong> 4), which revealed that<br />
pET25b (+) pelB leader sequence was not a suitable signal sequence to export this<br />
protease, in the set <strong>of</strong> conditions tested. In figures 3 <strong>and</strong> 4 representative results<br />
obtained for strain BL21(DE3) are shown for all the parameters tested (culture<br />
temperature, inducer concentration <strong>and</strong> induction phase temperature). Similar results<br />
were obtained for the strains BL21(DE3)pLysS <strong>and</strong> Tuner, using the same<br />
fermentations conditions.<br />
200.0<br />
116.5<br />
97.4<br />
66.2<br />
45.0<br />
31.0<br />
22.5<br />
14.3<br />
Osmotic shock I Osmotic shock II Soluble fraction Insoluble fraction<br />
MW 1 2 3 4 5 6 MW 7 8 9 10 11 12<br />
Figure 2. SDS-PAGE <strong>of</strong> proteins from E. coli BL21(DE3) cells grown at 37 ºC,<br />
induced <strong>with</strong> IPTG 1 mM at 30 ºC. Lanes 1, 4, 7 <strong>and</strong> 10: pET25pro-SubtilisinE. Lanes<br />
2, 5, 8 <strong>and</strong> 11: pET25-pro2SubtilisinE. Lanes 3, 6, 9 <strong>and</strong> 12: pET25-pro4SubtilisinE.<br />
MW: SDS-PAGE St<strong>and</strong>ard, Broad Range (Bio-Rad). The solid arrowheads indicate the<br />
position <strong>of</strong> recombinant proteins.<br />
189
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