Enzymatic degradation of poly(L-lactide) film by proteinase K: quartz crystal microbalance and atomic force microscopy study.
Enzymatic degradation of the poly(L-lactide) (PLLA) amorphous film by proteinase K has been investigated by combination of the complementary techniques of quartz crystal microbalance and atomic force microscopy (AFM). The erosion rate increased with increasing enzyme concentrations and attained to be constant under the condition of [proteinase K] > 100 microg/mL. The amount of the enzyme molecules adsorbed to the film was quantitatively evaluated at various concentrations by AFM, and it revealed that the erosion rate is determined by the amount of adsorbed enzyme. Adsorption of proteinase K was irreversible despite lack of the binding domain, so that the enzyme molecules on the film surface could be observed directly by AFM. Transformation of the enzyme molecule caused by packing in high density on the surface was observed at higher enzyme concentrations. The "footprint" of the individual proteinase K molecule on the PLLA film after enzymatic degradation suggests that the enzyme moves on the surface to hydrolyze the film around it.
Yamashita
Koichi
K
Advanced Development & Supporting Center, RIKEN Institute, 2-1 Hirosawa, Wako-shi, Saitama, 351-0198 Japan. koyama@riken.jp
Kikkawa
Yoshihiro
Y
Kurokawa
Kenji
K
Doi
Yoshiharu
Y
D000327:Adsorption; D001673:Biodegradation, Environmental; D019286:Endopeptidase K / Q000378:metabolism*; D007700:Kinetics; D018625:Microscopy, Atomic Force; D008970:Molecular Weight; D011091:Polyesters / Q000378:metabolism*; D011791:Quartz
D011091:Polyesters; D011791:Quartz; C033616:poly(lactide); D019286:Endopeptidase K