LAUSR

The sufficient structural thermostability of a biological macromolecule is an overriding need for green nanoreactors and nanofactories to secure high activity. However, little is still known about what specific structural motif is responsible for it. Here, graph theory was employed to examine if the temperature-dependent noncovalent interactions and metal bridges, as identified in the structures of Escherichia coli class II fructose 1,6-bisphosphate aldolase, could shape a systematic fluidic grid-like mesh network with topological grids to regulate the structural thermostability of the wild-type construct and its evolved variants in each generation upon decyclization. The results indicated that the biggest grids may govern the temperature thresholds for their tertiary structural perturbations but without affecting the catalytic activities. Moreover, lower grid-based systematic thermal instability may facilitate structural thermostability, but a highly independent thermostable grid may still be required to serve as a critical anchor to secure the stereospecific thermoactivity. Its end melting temperature thresholds, together with the start ones of the biggest grids in the evolved variants, may confer high temperature sensitivity against thermal inactivation. Collectively, this computational study may have widespread significance in advancing our complete understanding and biotechnology of the thermoadaptive mechanism of the structural thermostability of a biological macromolecule.