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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<strong>Surface</strong> <strong>Modification</strong> <strong>of</strong> <strong>Cellulose</strong> <strong>Acetate</strong> <strong>with</strong> <strong>Cutinase</strong> <strong>and</strong> <strong>Cutinase</strong> Fused to Carbohydrate-binding Modules<br />
CBM <strong>of</strong> Endoglucanase C (CenC) from Cellulomonas fimi, which belongs to the family<br />
CBM4, is able to bind amorphous cellulose (Boraston et al., 2005).<br />
To our knowledge, this is the first report <strong>of</strong> the hydrolysis <strong>of</strong> surface acetyl groups from<br />
CDA <strong>and</strong> CTA <strong>with</strong> a cutinase. It constitutes a promising approach for the partial<br />
regeneration <strong>of</strong> cellulose reactivity <strong>and</strong> hydrophilicity in these fibres, here demonstrated<br />
by the enhanced reactive dye uptake <strong>of</strong> treated fabrics. The production <strong>of</strong> fusion<br />
cutinases <strong>with</strong> new functionalities is here described <strong>and</strong> a comparison <strong>with</strong> cutinase<br />
regarding its efficiency for CDA <strong>and</strong> CTA modification is presented.<br />
2. Materials <strong>and</strong> methods<br />
2.1. Reagents <strong>and</strong> enzymes<br />
The cellulose diacetate <strong>and</strong> triacetate plain woven fabrics used were kindly supplied by<br />
Mitsubishi Rayon Co. Ltd., Tokyo, Japan. The CDA fabric has 41/27 ends/picks per cm<br />
<strong>and</strong> 64 g m -2 . The CTA fabric has 45/31 ends/picks per cm <strong>and</strong> 98 g m -2 .<br />
All other reagents were laboratory grade reagents from Sigma-Aldrich, St. Louis, USA,<br />
unless stated otherwise.<br />
The cutinase (EC 3.1.1.74) from F. solani pisi used in this work was expressed <strong>and</strong><br />
purified as previously reported by Araújo et al., 2007.<br />
Restriction enzymes were purchased from MBI Fermentas (Vilnius, Lithuania) <strong>and</strong><br />
from Roche Diagnostics GmbH (Penzberg, Germany). Accuzyme TM DNA polymerase<br />
was purchased from Bioline GmbH (Luckenwalde, Germany) <strong>and</strong> recombinant Taq<br />
DNA polymerase was purchased from MBI Fermentas. T4 DNA ligase was purchased<br />
from Roche Diagnostics GmbH (Penzberg, Germany).<br />
2.2. Esterase activity assay<br />
Esterase activity was determined following the product release (p-nitrophenol)<br />
continuously through the increase in the absorbance at 400 nm at 30º C. The assay<br />
conditions for the determination <strong>of</strong> cutinase activity were described previously (Matamá<br />
et al., 2006). All the assays were performed at least in triplicate. St<strong>and</strong>ard solutions <strong>of</strong><br />
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