The vast repertoire of carbohydrate oxidases:An overview

Carbohydrates are widely abundant molecules present in a variety of forms. For their biosynthesis and modification, nature has evolved a plethora of carbohydrate-acting enzymes. Many of these enzymes are of particular interest for biotechnological applications, where they can be used as biocatalysts or biosensors. Among the enzymes catalysing conversions of carbohydrates are the carbohydrate oxidases. These oxidative enzymes belong to different structural families and use different cofactors to perform the oxidation reaction of CH-OH bonds in carbohydrates. The variety of carbohydrate oxidases available in nature reflects their specificity towards different sugars and selectivity of the oxidation site. Thanks to their properties, carbohydrate oxidases have received a lot of attention in basic and applied research, such that nowadays their role in biotechnological processes is of paramount importance. In this review we provide an overview of the available knowledge concerning the known carbohydrate oxidases. The oxidases are first classified according to their structural features. After a description on their mechanism of action, substrate acceptance and characterisation, we report on the engineering of the different carbohydrate oxidases to enhance their employment in biocatalysis and biotechnology. In the last part of the review we highlight some practical applications for which such enzymes have been exploited

Exploring Deep Learning for In-Field Fault Detection in Microprocessors

Nowadays, due to technology enhancement, faults are increasingly compromising all kinds of computing machines, from servers to embedded systems. Recent advances in ma- chine learning are opening new opportunities to achieve fault detection exploiting hardware metrics inspection, thus avoiding the use of heavy software techniques or product-specific errors reporting mechanisms. This paper investigates the capability of different deep learning models trained on data collected through simulation-based fault injection to generalize over different software applications

Motorcycles that see: Multifocal stereo vision sensor for advanced safety systems in tilting vehicles

Advanced driver assistance systems, ADAS, have shown the possibility to anticipate crash accidents and effectively assist road users in critical traffic situations. This is not the case for motorcyclists, in fact ADAS for motorcycles are still barely developed. Our aim was to study a camera-based sensor for the application of preventive safety in tilting vehicles. We identified two road conflict situations for which automotive remote sensors installed in a tilting vehicle are likely to fail in the identification of critical obstacles. Accordingly, we set two experiments conducted in real traffic conditions to test our stereo vision sensor. Our promising results support the application of this type of sensors for advanced motorcycle safety applications

Mechanistic and Crystallographic Studies of Azoreductase AzoA from Bacillus wakoensis A01

The azoreductase AzoA from the alkali-tolerant Bacillus wakoensis A01 has been studied to reveal its structural and mechanistic details. For this, a recombinant expression system was developed which yields impressive amounts of fully active enzyme. The purified holo enzyme is remarkably solvent-tolerant and thermostable with an apparent melting temperature of 71 °C. The dimeric enzyme contains FMN as a prosthetic group and is strictly NADH dependent. While AzoA shows a negligible ability to use molecular oxygen as an electron acceptor, it is efficient in reducing various azo dyes and quinones. The kinetic and catalytic mechanism has been studied in detail using steady state kinetic analyses and stopped-flow studies. The data show that AzoA performs quinone and azo dye reductions via a two-electron transfer. Moreover, quinones were shown to be much better substrates (kcat values of 100-400 s-1 for several naphtoquinones) when compared with azo dyes. This suggests that the physiological role of AzoA and sequence-related microbial reductases is linked to quinone reductions and that they can better be annotated as quinone reductases. The structure of AzoA has been determined in complex with FMN at 1.8 Å resolution. AzoA displays unique features in the active site providing clues for explaining its catalytic and thermostability features. An uncommon loop, when compared with sequence-related reductases, forms an active site lid with Trp60 acting as an anchor. Several Trp60 mutants have been analyzed disclosing an important role of this residue in the stability of AzoA, while they retained activity. Structural details are discussed in relation to other azo and quinone reductases. This study provides new insights into the molecular functioning of AzoA and sequence-related reductases