Enzyme production of d-gluconic acid and glucose oxidase: successful tales of cascade reactions

This review mainly focuses on the use of glucose oxidase in the production of D-gluconic acid, which is a reactant of undoubtable interest in different industrial areas. The enzyme has been used in numerous instances as a model reaction to study the problems of oxygen supply in bioreactors. One of the main topics in this review is the problem of the generated side product, hydrogen peroxide, as it is an enzyme-inactivating reagent. Different ways to remove hydrogen peroxide have been used, such as metal catalysts and use of whole cells; however, the preferred method is the coupling glucose oxidase with catalase. The different possibilities of combining these enzymes have been discussed (use of free enzymes, independently immobilized enzymes or co-immobilized enzymes). Curiously, some studies propose the addition of hydrogen peroxide to this co-immobilized enzyme system to produce oxygen in situ. Other cascade reactions directed toward the production of gluconic acid from polymeric substrates will be presented; these will mainly involve the transformation of polysaccharides (amylases, cellulases, etc.) but will not be limited to those (e.g., gluconolactonase). In fact, glucose oxidase is perhaps one of most successful enzymes, and it is involved in a wide range of cascade reactions. Finally, other applications of the enzyme have been reviewed, always based on the production of D-gluconic acid, which produces a decrease in the pH, a decrease in the oxygen availability or the production of hydrogen peroxide; in many instances, cascade reactions are also utilized. Thus, this review presents many different cascade reactions and discusses the advantages/drawbacks of the use of co-immobilized enzymes.We gratefully recognize the financial support from Ministerio de Ciencia e Innovación-Spanish Government and FEDER funds (project number CTQ2017-86170-R, RTI2018-095291-BI00, MAT2017-87579-R) and Generalitat Valenciana (PROMETEO/2018/076). DC thank to Ministerio de Ciencia e Innovacion-Spanish Government by a FPI. PWT thanks to the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES) – Finance Code 001

Combi-CLEAs of glucose oxidase and catalase for conversion of glucose to gluconic acid eliminating the hydrogen peroxide to maintain enzyme activity in a bubble column reactor

© The Author(s).In this study combined cross-linked aggregates of catalase from bovine liver and glucose-oxidase from Aspergillus niger were prepared, and the effects of the precipitant and crosslinking agents, as well as the use of bovine serum albumin (BSA) as a feeder protein, on enzyme immobilization yield and thermal stability of both enzymes, were evaluated. Combi- crosslinking of enzyme aggregates (CLEAs) prepared using dimethoxyethane as precipitant, 25 mM glutaraldehyde and BSA/enzymes mass ratio of 5.45 (w/w), exhibited the highest enzyme activities and stabilities at 40 °C, pH 6.0, and 250 rpm for 5 h. The stability of both immobilized enzymes was fairly similar, eliminating one of the problems of enzyme coimmobilization. Combi-CLEAs were used in gluconic acid (GA) production in a bubble column reactor operated at 40 °C, pH 6.0 and 10 vvm of aeration, using 26 g L−1 glucose as the substrate. Results showed conversion of around 96% and a reaction course very similar to the same process using free enzymes. The operational half-life was 34 h, determined from kinetic profiles and the first order inactivation model. Combi-CLEAs of glucose-oxidase and catalase were shown to be a robust biocatalyst for applications in the production of gluconic acid from glucose.This work was supported by the National Council for Scientific and Technological Development (CNPq) (grants #141647/2013-2 and #402850/2013-0), and in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior–Brazil (CAPES)–Finance Code 001, and by the Comunidad Autónoma de Madrid (project Ref. IND2017/IND-7640) and MINECO from Spanish Government, (project number CTQ2017-86170-R).Peer reviewe

Effect of Concentrated Salts Solutions on the Stability of Immobilized Enzymes: Influence of Inactivation Conditions and Immobilization Protocol

This paper aims to investigate the effects of some salts (NaCl, (NH4)2SO4 and Na2SO4) at pH 5.0, 7.0 and 9.0 on the stability of 13 different immobilized enzymes: five lipases, three proteases, two glycosidases, and one laccase, penicillin G acylase and catalase. The enzymes were immobilized to prevent their aggregation. Lipases were immobilized via interfacial activation on octyl agarose or on glutaraldehyde-amino agarose beads, proteases on glyoxyl agarose or glutaraldehyde-amino agarose beads. The use of high concentrations of salts usually has some effects on enzyme stability, but the intensity and nature of these effects depends on the inactivation pH, nature and concentration of the salt, enzyme and immobilization protocol. The same salt can be a stabilizing or a destabilizing agent for a specific enzyme depending on its concentration, inactivation pH and immobilization protocol. Using lipases, (NH4)2SO4 generally permits the highest stabilities (although this is not a universal rule), but using the other enzymes this salt is in many instances a destabilizing agent. At pH 9.0, it is more likely to find a salt destabilizing effect than at pH 7.0. Results confirm the difficulty of foreseeing the effect of high concentrations of salts in a specific immobilized enzyme

Performance of a proposed event-type based analysis for the Cherenkov Telescope Array

The Cherenkov Telescope Array (CTA) will be the next-generation observatory in the field of very-high-energy (20 GeV to 300 TeV) gamma-ray astroparticle physics. Classically, data analysis in the field maximizes sensitivity by applying quality cuts on the data acquired. These cuts, optimized using Monte Carlo simulations, select higher quality events from the initial dataset. Subsequent steps of the analysis typically use the surviving events to calculate one set of instrument response functions (IRFs). An alternative approach is the use of event types, as implemented in experiments such as the Fermi-LAT. In this approach, events are divided into sub-samples based on their reconstruction quality, and a set of IRFs is calculated for each sub-sample. The sub-samples are then combined in a joint analysis, treating them as independent observations. This leads to an improvement in performance parameters such as sensitivity, angular and energy resolution. Data loss is reduced since lower quality events are included in the analysis as well, rather than discarded. In this study, machine learning methods will be used to classify events according to their expected angular reconstruction quality. We will report the impact on CTA high-level performance when applying such an event-type classification, compared to the classical procedure

Chasing Gravitational Waves with the Chereknov Telescope Array

Presented at the 38th International Cosmic Ray Conference (ICRC 2023), 2023 (arXiv:2309.08219)2310.07413International audienceThe detection of gravitational waves from a binary neutron star merger by Advanced LIGO and Advanced Virgo (GW170817), along with the discovery of the electromagnetic counterparts of this gravitational wave event, ushered in a new era of multimessenger astronomy, providing the first direct evidence that BNS mergers are progenitors of short gamma-ray bursts (GRBs). Such events may also produce very-high-energy (VHE, > 100GeV) photons which have yet to be detected in coincidence with a gravitational wave signal. The Cherenkov Telescope Array (CTA) is a next-generation VHE observatory which aims to be indispensable in this search, with an unparalleled sensitivity and ability to slew anywhere on the sky within a few tens of seconds. New observing modes and follow-up strategies are being developed for CTA to rapidly cover localization areas of gravitational wave events that are typically larger than the CTA field of view. This work will evaluate and provide estimations on the expected number of of gravitational wave events that will be observable with CTA, considering both on- and off-axis emission. In addition, we will present and discuss the prospects of potential follow-up strategies with CTA

Sensitivity of the Cherenkov Telescope Array to the gamma-ray emission from neutrino sources detected by IceCube

Gamma-ray observations of the astrophysical neutrino sources are fundamentally important for understanding the underlying neutrino production mechanism. We investigate the Cherenkov Telescope Array (CTA) ability to detect the very-high-energy (VHE) gamma-ray counterparts to the neutrino-emitting Active Galaxies. The CTA performance under different configurations and array layouts is computed based on the neutrino and gamma-ray simulations of steady and transient types of sources, assuming that the neutrino events are detected with the IceCube neutrino telescope. The CTA detection probability is calculated for both CTA sites taking into account the visibility constraints. We find that, under optimal observing conditions, CTA could observe the VHE gamma-ray emission from at least 3 neutrino events per year

Chasing Gravitational Waves with the Chereknov Telescope Array

Presented at the 38th International Cosmic Ray Conference (ICRC 2023), 2023 (arXiv:2309.08219)2310.07413International audienceThe detection of gravitational waves from a binary neutron star merger by Advanced LIGO and Advanced Virgo (GW170817), along with the discovery of the electromagnetic counterparts of this gravitational wave event, ushered in a new era of multimessenger astronomy, providing the first direct evidence that BNS mergers are progenitors of short gamma-ray bursts (GRBs). Such events may also produce very-high-energy (VHE, > 100GeV) photons which have yet to be detected in coincidence with a gravitational wave signal. The Cherenkov Telescope Array (CTA) is a next-generation VHE observatory which aims to be indispensable in this search, with an unparalleled sensitivity and ability to slew anywhere on the sky within a few tens of seconds. New observing modes and follow-up strategies are being developed for CTA to rapidly cover localization areas of gravitational wave events that are typically larger than the CTA field of view. This work will evaluate and provide estimations on the expected number of of gravitational wave events that will be observable with CTA, considering both on- and off-axis emission. In addition, we will present and discuss the prospects of potential follow-up strategies with CTA

Chasing Gravitational Waves with the Chereknov Telescope Array

Presented at the 38th International Cosmic Ray Conference (ICRC 2023), 2023 (arXiv:2309.08219)2310.07413International audienceThe detection of gravitational waves from a binary neutron star merger by Advanced LIGO and Advanced Virgo (GW170817), along with the discovery of the electromagnetic counterparts of this gravitational wave event, ushered in a new era of multimessenger astronomy, providing the first direct evidence that BNS mergers are progenitors of short gamma-ray bursts (GRBs). Such events may also produce very-high-energy (VHE, > 100GeV) photons which have yet to be detected in coincidence with a gravitational wave signal. The Cherenkov Telescope Array (CTA) is a next-generation VHE observatory which aims to be indispensable in this search, with an unparalleled sensitivity and ability to slew anywhere on the sky within a few tens of seconds. New observing modes and follow-up strategies are being developed for CTA to rapidly cover localization areas of gravitational wave events that are typically larger than the CTA field of view. This work will evaluate and provide estimations on the expected number of of gravitational wave events that will be observable with CTA, considering both on- and off-axis emission. In addition, we will present and discuss the prospects of potential follow-up strategies with CTA

Interpolation of Instrument Response Functions for the Cherenkov Telescope Array in the Context of pyirf

The Cherenkov Telescope Array (CTA) will be the next generation ground-basedvery-high-energy gamma-ray observatory, constituted by tens of Imaging AtmosphericCherenkov Telescopes at two sites once its construction and commissioning are finished. Like its predecessors, CTA relies on Instrument Response Functions (IRFs) to relate the observed and reconstructed properties to the true ones of the primary gamma-ray photons. IRFs are needed for the proper reconstruction of spectral and spatial information of the observed sources and are thus among the data products issued to the observatory users. They are derived from Monte Carlo simulations, depend on observation conditions likethe telescope pointing direction or the atmospheric transparency and can evolve with time as hardware ages or is replaced. Producing a complete set of IRFs from simulations for every observation taken is a time-consuming task and not feasible when releasing data products on short timescales. Consequently, interpolation techniques on simulated IRFs are investigated to quickly estimate IRFs for specific observation conditions. However, as some of the IRFs constituents are given as probability distributions, specialized methods are needed. This contribution summarizes and compares the feasibility of multiple approaches to interpolate IRF components in the context of the pyirf python software package and IRFs simulated for the Large-Sized Telescope prototype (LST-1). We will also give an overview of the current functionalities implemented in pyirf

Variability studies of active galactic nuclei from the long-term monitoring program with the Cherenkov Telescope Array

Blazars are active galactic nuclei (AGN) with a relativistic jet oriented toward the observer. This jet is composed of accelerated particles which can display emission over the entire electromagnetic spectrum. Spectral variability has been observed on short- and long-time scales in AGN, with a power spectral density (PSD) that can show a break at frequencies below the well-known red-noise process. This break frequency in the PSD has been observed in X-rays to scale with the accretion regime and the mass of the central black hole. It is expected that a break could also be seen in the very-high-energy gamma rays, but constraining the shape of the PSD in these wavelengths has not been possible with the current instruments. The Cherenkov Telescope Array (CTA) will be more sensitive by a factor of five to ten depending on energy than the current generation of imaging atmospheric Cherenkov telescopes, therefore it will be possible with CTA to reconstruct the PSD with a high accuracy, bringing new information about AGN variability. In this work, we focus on the AGN long-term monitoring program planned with CTA. The program is proposed to begin with early-start observing campaigns with CTA precursors. This would allow us to probe longer time scales on the AGN PSD