2.9. Reconstruction experiment to reproduce the secretion and immobilization of environmental cellulase

Autoclaved reed leaves/cellulose were immersed inside a sieve (45 µm mesh, ϕ = 75 mm) for easy collection in a culture tank containing 2 L AAS and 25 C. japonica individuals. After 24 hr, leaves (1mg, 100 µl CAMS, n = 3) and cellulose (25 mg/duplicate, 100 µl CAMS, n = 5) were collected and washed with 1 ml AAS 5 times before measuring reducing sugar production or glucose production. For Western blotting and immunofluorescence, 5 mg cellulose was used for each assay. Alexa Fluor 546 (Life Technologies) was used as the secondary antibody for immunofluorescence.