GH32 family activity: a topological approach through protein contact networks.

KEY MESSAGE: The application of Protein Contact Networks methodology allowed to highlight a novel response of border region between the two domains to substrate binding. Glycoside hydrolases (GH) are enzymes that mainly hydrolyze the glycosidic bond between two carbohydrates or a carbohydrate and a non-carbohydrate moiety. These enzymes are involved in many fundamental and diverse biological processes in plants. We have focused on the GH32 family, including enzymes very similar in both sequence and structure, each having however clear specificities of substrate preferences and kinetic properties. Structural and topological differences among proteins of the GH32 family have been here identified by means of an emerging approach (Protein Contact network, PCN) based on the formalization of 3D structures as contact networks among amino-acid residues. The PCN approach proved successful in both reconstructing the already known functional domains and in identifying the structural counterpart of the properties of GH32 enzymes, which remain uncertain, like their allosteric character. The main outcome of the study was the discovery of the activation upon binding of the border (cleft) region between the two domains. This reveals the allosteric nature of the enzymatic activity for all the analyzed forms in the GH32 family, a character yet to be highlighted in biochemical studies. Furthermore, we have been able to recognize a topological signature (graph energy) of the different affinity of the enzymes towards small and large substrates.

Sludge reduction in a small wastewater treatment plant by electro-kinetic disintegration.

Sludge reduction in a wastewater treatment plant (WWTP) has recently become a key issue for the managing companies, due to the increasing constraints on the disposal alternatives. Therefore, all the solutions proposed with the aim of minimizing sludge production are receiving increasing attention and are tested either at laboratory or full-scale to evaluate their real effectiveness. In the present paper, electro-kinetic disintegration has been applied at full-scale in the recycle loop of the sludge drawn from the secondary settlement tank of a small WWTP for domestic sewage. After the disintegration stage, the treated sludge was returned to the biological reactor. Three different percentages (50, 75 and 100%) of the return sludge flow rate were subjected to disintegration and the effects on the sludge production and the WWTP operation efficiency evaluated. The long-term observations showed that the electro-kinetic disintegration was able to drastically reduce the amount of biological sludge produced by the plant, without affecting its treatment efficiency. The highest reduction was achieved when 100% return sludge flow rate was subjected to the disintegration process. The reduced sludge production gave rise to a considerable net cost saving for the company which manages the plant.