TY - JOUR
T1 - Tailoring cutinase activity towards polyethylene terephthalate and polyamide 6,6 fibers
AU - Araújo, Rita
AU - Silva, Carla
AU - O'Neill, Alexandre
AU - Micaelo, Nuno
AU - Guebitz, Georg
AU - Soares, Cláudio M.
AU - Casal, Margarida
AU - Cavaco-Paulo, Artur
PY - 2007/3/10
Y1 - 2007/3/10
N2 - Cutinase from Fusarium solani pisi was genetically modified near the active site, by site-directed mutagenesis, to enhance its activity towards polyethylene terephthalate (PET) and polyamide 6,6 (PA 6,6) fibers. The mutations L81A, N84A, L182A, V184A and L189A were done to enlarge the active site in order to better fit a larger polymer chain. Modeling studies have shown enhanced free energy stabilization of model substrate tetrahedral intermediate (TI) bound at the enzyme active site for all mutants, for both model polymers. L81A and L182A showed an activity increase of four- and five-fold, respectively, when compared with the wild type, for PET fibers. L182A showed the one- and two-fold higher ability to biodegrade aliphatic polyamide substrates. Further studies in aliphatic polyesters seem to indicate that cutinase has higher ability to recognize aliphatic substrates.
AB - Cutinase from Fusarium solani pisi was genetically modified near the active site, by site-directed mutagenesis, to enhance its activity towards polyethylene terephthalate (PET) and polyamide 6,6 (PA 6,6) fibers. The mutations L81A, N84A, L182A, V184A and L189A were done to enlarge the active site in order to better fit a larger polymer chain. Modeling studies have shown enhanced free energy stabilization of model substrate tetrahedral intermediate (TI) bound at the enzyme active site for all mutants, for both model polymers. L81A and L182A showed an activity increase of four- and five-fold, respectively, when compared with the wild type, for PET fibers. L182A showed the one- and two-fold higher ability to biodegrade aliphatic polyamide substrates. Further studies in aliphatic polyesters seem to indicate that cutinase has higher ability to recognize aliphatic substrates.
KW - Biocatalysis
KW - Cutinase
KW - Polyamide 6,6
KW - Polyester
KW - Site-directed mutagenesis
UR - http://www.scopus.com/inward/record.url?scp=33847353686&partnerID=8YFLogxK
U2 - 10.1016/j.jbiotec.2006.12.028
DO - 10.1016/j.jbiotec.2006.12.028
M3 - Article
C2 - 17306400
AN - SCOPUS:33847353686
SN - 0168-1656
VL - 128
SP - 849
EP - 857
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 4
ER -