Single molecule image sequences of Thermothielavioides terrestris AA9E (TtAA9E) on cellulose in oxygen scavenging systems

Purified and Cy5-labeled lytic polysaccharide monooxygenase (LPMO) Thermothielavioides terrestris AA9E (TtAA9E) interacting at the surface of crystalline cellulose fibrils were imaged by Total Internal Reflectance Fluorescence Microscopy (TIRFM) in two different oxygen scavenging systems: Glucose Oxidase-Catalase (GODCAT) and Protocatechuic Acid/Protocatechuate-3,4-dioxygenase (PCA/PCD). The datasets (movies) show how oxygen and hydrogen peroxide availability, and reductant type/availability impact specific binding of the LPMO to highly ordered cellulose fibrils., , , # Single Molecule Image Sequences of *Thermothielavioides terrestris* AA9E (*Tt*AA9E) on Cellulose in Oxygen Scavenging System

Dataset DOI: [10.5061/dryad.9ghx3ffrh](10.5061/dryad.9ghx3ffrh)

Single-molecule fluorescence images of individual Cy5-labeled enzyme molecules binding and unbinding from cellulose fibrils were imaged using 488 nm and 637 nm excitation laser reflected by a multiline dichroic mirror (FF500/646-Di01, Semrock) and focused at the back aperture of a 1.49 NA 60X oil-immersion objective (Olympus) to provide total-internal-reflection (TIR) excitation at the cover glass / water interface across a ~50 µm diameter area in the object plane. Sample emission was collected and imaged by the same objective onto the 512x512-pixel sensor of an electron multiplying CCD (EMCCD) camera (Photonmax, Princeton Instruments). A 37 nm wide bandpass filter centered at 676 nm was used to isolate Cy5 fluorescence excited at 637 nm. For the binding time measurements, the overall magnificatio...,