Installing hydrolytic activity into a completely <i>de novo </i>protein framework

The design of enzyme-like catalysts tests our understanding of sequence-to-structure/function relationships in proteins. Here we install hydrolytic activity predictably into a completely de novo and thermostable α-helical barrel, which comprises seven helices arranged around an accessible channel. We show that the lumen of the barrel accepts 21 mutations to functional polar residues. The resulting variant, which has cysteine–histidine–glutamic acid triads on each helix, hydrolyses p-nitrophenyl acetate with catalytic efficiencies that match the most-efficient redesigned hydrolases based on natural protein scaffolds. This is the first report of a functional catalytic triad engineered into a de novo protein framework. The flexibility of our system also allows the facile incorporation of unnatural side chains to improve activity and probe the catalytic mechanism. Such a predictable and robust construction of truly de novo biocatalysts holds promise for applications in chemical and biochemical synthesis

Human-in-Loop Decision-Making and Autonomy: Lessons Learnt from the Aviation Industry Transferred to Cyber-Physical Systems

Peer reviewed: TrueIn this study, we reviewed aircraft accidents in order to understand how autonomy and safety has been managed in the aviation industry, with the aim of transferring our findings to autonomous cyber-physical systems (CPSs) in general. Through the qualitative analysis of 26 reports of aircraft accidents that took place from 2016 to 2022, we identified the most common contributing factors and the actors involved in aircraft accidents. We found that accidents were rarely the result of a single event or actor, with the most common contributing factor being non-adherence to standard operating procedures (SOPs). Considering that the aviation industry has had decades to perfect their SOPs, it is important for CPSs not only to consider the actors and causes that may contribute to safety-related issues, but also to consider well-defined reporting practices, as well as the different levels of mechanisms checked by diverse stakeholders, in order to minimise the cascading nature of such events to improve safety. In addition to proposing a new definition of safety, in this study we suggest reviewing high-reliability organisations to offer further insights as part of future research on CPS safety.</jats:p