Using high-speed scanning atomic force microscopy, we directly observed single-molecular enzymatic elongation of hyaluronan polymer chains at intervals of 10 s on a mica or lipid bilayer surface, on which Pasteurella multocida hyaluronic acid synthase (pmHAS) was immobilized. The reaction was started by the addition of both UDP-glucuronic acid and UDP-N-acetylglucosamine monomers. The average catalytic elongation rate constant (kcat) was found to be 1.8 mer s-1 from one active enzyme physically adsorbed on a mica surface. When pmHAS was immobilized by inserting its hydrophobic tail part into lipid bilayers, most of the enzymes retained their activity, and the k cat values were found to be in the range 1-10 mer s-1 for 29 enzymes (average was kcat = 2-4 mer s-1). These k cat values were lowest level of kcat = 1-100 s -1 obtained in bulk solution by radioisotope methods.
|Number of pages||4|
|Journal||Journal of the American Chemical Society|
|Publication status||Published - Dec 19 2012|
All Science Journal Classification (ASJC) codes
- Colloid and Surface Chemistry