Abstract
Extremozymes have gained considerable interest as they could meet industrial requirements. Among these, SsoPox is a hyperthermostable enzyme isolated from the archaeon Sulfolobus solfataricus. This enzyme is a lactonase catalyzing the hydrolysis of acyl-homoserine lactones; these molecules are involved in Gram-negative bacterial communication referred to as quorum sensing. SsoPox exhibits promiscuous phosphotriesterase activity for the degradation of organophosphorous chemicals including insecticides and chemical warfare agents. Owing to its bi-functional catalytic abilities as well as its intrinsic stability, SsoPox is appealing for many applications, having potential uses in the agriculture, defense, food and health industries. Here we investigate the biotechnological properties of the mutant SsoPox-W263I, a variant with increased lactonase and phosphotriesterase activities. We tested enzyme resistance against diverse process-like and operating conditions such as heat resistance, contact with organic solvents, sterilization, storage and immobilization. Bacterial secreted materials from both Gram-negative and positive bacteria were harmless on SsoPox-W263I activity and could reactivate heat-inactivated enzyme. SsoPox showed resistance to harsh conditions demonstrating that it is an extremely attractive enzyme for many applications. Finally, the potential of SsoPox-W263I to be active at subzero temperature is highlighted and discussed in regards to the common idea that hyperthermophile enzymes are nearly inactive at low temperatures.
Highlights
To classical antimicrobial agents[24,25,26]
Our results show that SsoPox-W263I maintains most of its activity after extremely harsh treatments, highlighting the tremendous potential for using this enzyme in existing industrial production lines
We investigated the resistance of SsoPox-W263I submitted to a heat shock from 40 °C up to 120 °C in liquid form and 150 °C in solid form for 5 minutes using a dry bath to simulate reticulation step used for example in textile processes (Fig. 1)
Summary
To classical antimicrobial agents (e.g. antibiotics)[24,25,26]. Industrial applications using QQ strategy mainly include medical devices such as catheters, aerosols or dressings and have been exhaustively reviewed elsewhere[27,28]. OP were considered for military ends leading to the synthesis of noxious chemical warfare nerve agents (CWNA)[36,37] These compounds constitute a serious threat for civil and military populations but no satisfactory external decontamination method is currently available[38]. OP-degrading enzymes have emerged as potential bioremediation alternatives[38,39] Considering both its lactonase and phosphotriesterase capabilities as well as its exceptional thermal stability (Tm = 106 °C), many applications involving SsoPox are considered[38,40,41,42,43,44,45,46]. Variant SsoPox-W263I, which exhibits higher lactonase and phosphotriesterase activity, is a promising candidate for the external bioremediation of OP, including liquid decontamination solutions, filtration systems, and auto-decontaminating textiles or materials. Our results show that SsoPox-W263I maintains most of its activity after extremely harsh treatments, highlighting the tremendous potential for using this enzyme in existing industrial production lines
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