In this work, the
copolymer poly[(9,9-dioctylfluorene)-co-(3-hexylthiophene)]
was employed as a matrix for immobilizing
phytase, aiming at the detection of phytic acid. The copolymer was
spread on the air–water interface forming Langmuir monolayers
and phytase adsorbed from the aqueous subphase. The interactions between
the copolymer and the enzyme components were investigated with surface
pressure and surface potential–area isotherms, Brewster angle
microscopy, and polarization modulation infrared reflection–absorption
spectroscopy (PM-IRRAS). The enzyme could be incorporated in the monolayers
from the aqueous subphase, expanding the copolymer films and maintaining
its secondary structure. The polymeric films presented a morphological
heterogeneous pattern at the air–water interface because of
the ability of their chains to fold and entangle, causing inherent
defects in the organization as well as unbalanced lateral distribution
at the air–water interface because of the formation of aggregates.
The interfacial films were transferred to solid supports as Langmuir–Blodgett
films and characterized by PM-IRRAS and scanning electronic microscopy,
which showed not only the co-transfer of the enzyme but also the maintenance
of their heterogeneous morphological pattern. The enzymatic activity
of the blended film was analyzed by UV–vis spectroscopy and
allowed the estimation of the value of the Michaelis constant (13.08
mM), demonstrating the feasibility of the system to selectively detect
phytic acid for biosensing purposes.