Acyl-enzyme intermediate 2026-10-20
A covalent enzyme intermediate bearing an acyl group during catalysis. Its formation and breakdown can occur at different rates, producing burst kinetics if the first released reaction product reports a fast step.
A detectable burst kinetics phase requires rapid formation and detection of an initial reaction product, followed by a slower step that regenerates competent enzyme. A common hydrolysis mechanism is
where can be a covalent acyl-enzyme intermediate. With excess substrate, rapid binding and , the first turnover rapidly releases and traps much of the enzyme as . Subsequent turnovers wait for the slow regeneration step. The measured product must report the fast step: monitoring only need not reveal a burst. Fast mixing and sufficient time resolution are needed, while spontaneous substrate hydrolysis, depletion and enzyme inactivation must be excluded.
For an illustrative saturated two-step cycle, initially unmodified enzyme concentration gives
The second term approaches a burst amplitude near when , followed by the slower steady-state slope. A covalent intermediate is a useful example, not a logical requirement for every possible burst mechanism.