Thin films of lignocellulosic nanofibrils (LCNFs) with different chemical compositions were prepared for real-time observation of their enzymatic adsorption and degradation behavior by using a quartz crystal microbalance with dissipation monitoring (QCM-D). LCNFs were obtained by disk milling followed by high-pressure homogenization of Hinoki cypress. The lignin contents were adjusted by the sodium chlorite treatment. The film thickness was adjusted by controlling the concentration of the LCNF suspension, which was determined from its proportional relationship to the UV absorbance of lignin. The enzymatic degradation behavior was investigated with a commercial enzyme mixture. The results of the QCM-D showed that changes in frequency and dissipation in the initial reaction stage were different from the typical changes reported for pure cellulose. To the best of our knowledge, this is the first report of the preparation of thin films of LCNFs with high lignin and hemicellulose contents and their application in a QCM-D study.