TY - JOUR
T1 - Improving enzymatic polyurethane hydrolysis by tuning enzyme sorption
AU - Gamerith, Caroline
AU - Herrero Acero, Enrique
AU - Pellis, Alessandro
AU - Ortner, Andreas
AU - Vielnascher, Robert
AU - Luschnig, Daniel
AU - Zartl, Barbara
AU - Haernvall, Karolina
AU - Zitzenbacher, Sabine
AU - Strohmeier, Gernot
AU - Hoff, Oskar
AU - Steinkellner, Georg
AU - Gruber, Karl
AU - Ribitsch, Doris
AU - Guebitz, Georg M.
PY - 2016/3/4
Y1 - 2016/3/4
N2 - In this study we investigated the ability of amidases to hydrolyse polyurethane polyester co-polymers. In order to improve enzyme adsorption, a polyamidase from Nocardia farcinica (PA) was fused to a polymer binding module from a polyhydroxyalkanoate depolymerase from Alcaligenes faecalis (PA_PBM). The activity of these enzymes and of various commercially available amidases on a synthesized soluble model substrate was compared. The recombinant native PA showed the highest activity of 10.5 U/mg followed by PA_PBM with an activity of 1.13 U/mg. Both enzymes were able to cleave the urethane bond in polyurethane-polyesters with different degree of crystallinity as shown by FTIR. According to LC-TOF analysis the monomer 4,4′-diaminodiphenylmethane (MDA) and the oligomers 4-hydroxybutyl (3-(3-aminobenzyl)phenyl)carbamate, bis(4-hydroxybutyl) (methylenebis(3,1-phenylene))dicarbamate and 4-(((3-(3-(((4-hydroxybutoxy)carbonyl)amino)benzyl)phenyl)carbamoyl)oxy)butyl (4-hydroxybutyl) adipate were released. The polymer with a higher content of the rigid segment, MDA, was hydrolysed to a lower extent. Interestingly, despite the lower activity on the soluble model substrate, the PA_PBM fusion enzyme was up to 4 times more active on the polymer when compared with the native enzyme, confirming the relevance of enzyme adsorption for efficient hydrolysis.
AB - In this study we investigated the ability of amidases to hydrolyse polyurethane polyester co-polymers. In order to improve enzyme adsorption, a polyamidase from Nocardia farcinica (PA) was fused to a polymer binding module from a polyhydroxyalkanoate depolymerase from Alcaligenes faecalis (PA_PBM). The activity of these enzymes and of various commercially available amidases on a synthesized soluble model substrate was compared. The recombinant native PA showed the highest activity of 10.5 U/mg followed by PA_PBM with an activity of 1.13 U/mg. Both enzymes were able to cleave the urethane bond in polyurethane-polyesters with different degree of crystallinity as shown by FTIR. According to LC-TOF analysis the monomer 4,4′-diaminodiphenylmethane (MDA) and the oligomers 4-hydroxybutyl (3-(3-aminobenzyl)phenyl)carbamate, bis(4-hydroxybutyl) (methylenebis(3,1-phenylene))dicarbamate and 4-(((3-(3-(((4-hydroxybutoxy)carbonyl)amino)benzyl)phenyl)carbamoyl)oxy)butyl (4-hydroxybutyl) adipate were released. The polymer with a higher content of the rigid segment, MDA, was hydrolysed to a lower extent. Interestingly, despite the lower activity on the soluble model substrate, the PA_PBM fusion enzyme was up to 4 times more active on the polymer when compared with the native enzyme, confirming the relevance of enzyme adsorption for efficient hydrolysis.
KW - Polyurethane
KW - Polyamidase
KW - Enzymatic degradation
KW - Polyurethane model substrate
KW - Functionalization
U2 - 10.1016/j.polymdegradstab.2016.02.025
DO - 10.1016/j.polymdegradstab.2016.02.025
M3 - Article
VL - 132
SP - 69
EP - 77
JO - Polymer degradation and stability
JF - Polymer degradation and stability
ER -