Xylanase Production By Thermobacillus Xylanilyticus is Impaired By Population Diversification But Can Be Mitigated Based On the Management of Cheating Behavior

Background: The microbial production of hemicellulasic cocktails is still a challenge for the sector of biorefineries and agro-waste valorization. In this work, the production of hemicellulolytic enzymes by Thermobacillus xylanilyticus has been considered. This microorganism is of interest since it is able to produce an original set of thermostable hemicellulolytic enzymes, and notably a xylanase GH11, Tx-xyn11. However, cell-to-cell heterogeneities impairs the production capability of the whole microbial population.Results: Sequential cultivations of the strain on xylan as a carbon source has been considered in order to highlight and better understand this cell-to-cell heterogeneity. Successive cultivations pointed out a fast decrease of xylanase activity (loss of ~75%) after 23.5 generations. Accordingly, the expression of the Tx-xyn11 gene decreased drastically and followed the same trend as the xylanase activity. Flow cytometry analyses pointed out that two subpopulations, differing at the level of their light scattering properties, were potentially involved in this progressive loss of enzymatic activities. Interestingly, upon successive cultivations on xylan, the subpopulation exhibiting low forward scatter (FSC) signal. Additionally, the evolution of the ratio between the two subpopulations was correlated to the decrease in xylanase activity. Cell sorting and direct observation of the sorted subpopulations revealed that the low-FSC subpopulation was not sporulating, whereas the high-FSC subpopulation contained cells at the onset of the sporulation stage. Serial cultivations on glucose, followed by the addition of a xylan pulse led to a ~1.5-fold to ~15-fold improvement of xylanase, depending on the moment for pulse addition, , suggesting that alternating cultivation conditions could lead to an efficient population management strategy for the production of xylanase. Conclusions: Taken altogether, the data from this study point out that a cheating behaviour is responsible for the progressive reduction in xylanase activity during serial cultivations of T. xylanilyticus. Alternating cultivation condition between glucose and xylan could be used as an efficient strategy for promoting population stability and higher enzymatic productivity from this bacterium.

Superior operational stability of immobilized l-asparaginase over surface-modified carbon nanotubes

Abstractl-asparaginase (ASNase, EC 3.5.1.1) is an enzyme that catalyzes the l-asparagine hydrolysis into l-aspartic acid and ammonia, being mainly applied in pharmaceutical and food industries. However, some disadvantages are associated with its free form, such as the ASNase short half-life, which may be overcome by enzyme immobilization. In this work, the immobilization of ASNase by adsorption over pristine and modified multi-walled carbon nanotubes (MWCNTs) was investigated, the latter corresponding to functionalized MWCNTs through a hydrothermal oxidation treatment. Different operating conditions, including pH, contact time and ASNase/MWCNT mass ratio, as well as the operational stability of the immobilized ASNase, were evaluated. For comparison purposes, data regarding the ASNase immobilization with pristine MWCNT was detailed. The characterization of the ASNase-MWCNT bioconjugate was addressed using different techniques, namely Transmission Electron Microscopy (TEM), Thermogravimetric Analysis (TGA) and Raman spectroscopy. Functionalized MWCNTs showed promising results, with an immobilization yield and a relative recovered activity of commercial ASNase above 95% under the optimized adsorption conditions (pH 8, 60 min of contact and 1.5 × 10–3 g mL−1 of ASNase). The ASNase-MWCNT bioconjugate also showed improved enzyme operational stability (6 consecutive reaction cycles without activity loss), paving the way for its use in industrial processes.

Bimetallic MOF-Derived Synthesis of Cobalt-Cerium Oxide Supported Phosphotungstic Acid Composites for the Oleic Acid Esterification

The impregnation of phosphotungstic acid (HPW) with porous cobalt-cerium oxide (HPW@CoCeO) has been prepared by pyrolysis of CoCe-MOF and used for the production of methyl oleate from oleic acid and methanol. FTIR, XRD, SEM, TEM, N2 adsorption/desorption, and NH3-TPD were characterized for the prepared composites. Simultaneously, the effects of reaction time, substrate molar ratio, temperature, and catalyst loading on catalytic activity were highlighted, and the conversion of 67.2% was reached after 4 h at 60°C. Importantly, HPW@CoCeO was reusabe and reused more than eight times, and the oleic acid conversion could be maintained at 61.8% without significant activity loss. Thus, the HPW@CoCeO composite could be used as acid catalysts for sustainable energy production.

Hybrid Structure of TiO2-Graphitic Carbon as a Support of Pt Nanoparticles for Catalyzing Oxygen Reduction Reaction

The durability of catalysts in fuel cells is a longstanding issue that needs to be resolved. Catalyst stability of the fuel cell has always been a problem, studies are underway to address them. Herein, to address this issue, we synthesize a hybrid structure consisting of SP carbon (SP) as the graphitic carbon and TiO2 as the metal oxide using a microwave method for use as a support for Pt nanoparticles. Anatase TiO2 and Pt nanoparticles with sizes of ~5 and 3.5 ± 1.4 nm, respectively, are uniformly dispersed on a modified graphitic SP carbon support (Pt-TiO2-SP). This supported Pt catalyst exhibits significantly improves durability in the oxygen reduction reaction (ORR). Furthermore, the Pt-TiO2-SP carbon hybrid catalyst manifests superior electrocatalytic stability and higher onset potential in ORR than those exhibited by Pt-SP carbon without TiO2. Pt-TiO2-SP exhibits an activity loss of less than 68 mV after 5000 electrochemical cycles, whereas an activity loss of ~100 mV is observed for Pt-SP carbon in a stability test. These results suggest that the strong metal–support interaction in TiO2-supported Pt catalyst significantly enhances the activity of Pt nanocatalyst.

Functioned catalysts with magnetic core applied in the ibuprofen degradation

In the present work, the performance of Ag/ZnO/CoFe2O4 magnetic photocatalysts in the photocatalytic degradation of Ibuprofen (IBP) was evaluated. This study considered the use of pure Ag/ZnO (5% Ag) and also use the Ag/ZnO/CoFe2O4 magnetic catalysts containing different amounts (5, 10 and 15% wt) of cobalt ferrite (CoFe2O4). The catalysts were characterized by Scanning Electron Microscopy (SEM), X-ray diffraction (XRD) and photoacoustic Spectroscopy. To carried the photocatalytic degradation reaction, different concentrations of the ibuprofen contaminant solution (10, 20 and 30 ppm) and different concentrations of photocatalyst were tested (0.3 gL−1, 0.5 gL−1 and 1.0 gL−1). The reaction parameters studied were: IBP concentration, catalyst concentration, adsorption and photolysis, influence of the matrix, radiation source (solar and artificial) and the effect of organic additive. At the end of the photocatalytic tests, the best operation conditions were defined. Considering the obtained results of degradation efficiency and magnetic separation, the optimal parameters selected to proceed with the other tests of the study were: ibuprofen solution concentration 10 ppm, Ag/ZnO/CoFe2O4(5%) catalyst at a concentration of 0.3 g L−1 and pH 4.5 of the reaction medium. The results indicated the feasibility of magnetic separation of the synthesized catalysts. A long duration test indicated that the catalyst exhibits stability throughout the degradation reaction, as more than 80% of ibuprofen was degraded after 300 minutes. The photocatalytic activity was directly affected by the ferrite load. The higher the nominal load of ferrite, the lower the performance in ibuprofen degrading. It was also observed that the smallest amount of ferrite studied was enough for the catalyst to be recovered and reused. The adsorption and photolysis tests did not show significant results in the IBP degradation. In addition, it was possible to verify that the aqueous matrix, the use of solar radiation and the addition of additive (acid formic) interfered direct in the process. The catalyst reuse tests indicated that it can be recovered and reused at least three times without considerable catalytic activity loss.

Aza-Reversine Promotes Reprogramming of Lung (MRC-5) and Differentiation of Mesenchymal Cells into Osteoblasts

Reversine or 2-(4-morpholinoanilino)-N6-cyclohexyladenine was originally identified as a small organic molecule that induces dedifferentiation of lineage-committed mouse myoblasts, C2C12, and redirects them into lipocytes or osteoblasts under lineage-specific conditions (LISCs). Further, it was proven that this small molecule can induce cell cycle arrest and apoptosis and thus selectively lead cancer cells to cell death. Further studies demonstrated that reversine, and more specifically the C2 position of the purine ring, can tolerate a wide range of substitutions without activity loss. In this study, a piperazine analog of reversine, also known as aza-reversine, and a biotinylated derivative of aza-reversine were synthesized, and their potential medical applications were investigated by transforming the endoderm originates fetal lung cells (MRC-5) into the mesoderm originated osteoblasts and by differentiating mesenchymal cells into osteoblasts. Moreover, the reprogramming capacity of aza-reversine and biotinylated aza-reversine was investigated against MRC-5 cells and mesenchymal cells after the immobilization on PMMA/HEMA polymeric surfaces. The results showed that both aza-reversine and the biofunctionalized, biotinylated analog induced the reprogramming of MRC-5 cells to a more primitive, pluripotent state and can further transform them into osteoblasts under osteogenic culture conditions. These molecules also induced the differentiation of dental and adipose mesenchymal cells to osteoblasts. Thus, the possibility to load a small molecule with useful “information” for delivering that into specific cell targets opens new therapeutic personalized applications.

Frailty in Stroke; a Narrated Review

This narrative review provides a summary introduction to the relationship between stroke and physical and cognitive frailty syndromes and the neuro-inflammatory similarities (including inflammaging) between the two. The review argues the potential effects of Post COVID-19 Neurological Syndrome (PCNS, also known as Long COVID) with similar pathophysiology. Many patients who have suffered from acute stroke experience long-lasting symptoms affecting several organs including fatigue, brain fog, reduced physical activity, loss of energy, and loss of cognitive reserve, culminating in the loss of independence and poor quality of life. This is very similar to the emerging reports of PCNS from different parts of the world. Stroke, particularly in older adults with comorbidities appears to impact the health and welfare of patients by reducing central neuronal input and neuromuscular function, with muscular atrophy and neuropsychiatric complications. The cumulative effects can potentially lead to a range of physical and cognitive frailty syndromes, which, in many cases may be attributed to persistent, maladapted, low grade, chronic inflammation. Meanwhile, post-COVID-19 Neurological Syndrome (also known as Long COVID Syndrome) appears to share a similar trajectory, adding further urgency for investigations into the mechanisms underlying this constellation of symptoms.

Facile preparation and photocatalytic performance of anatase TiO2/nanocellulose composite

Anatase TiO2/nanocellulose composite was prepared for the first time via a one-step method at a relatively low temperature by using cellulose nanofibers as carrier and tetrabutyl titanate as titanium precursor. The morphology, structure and element composition of the composite were characterized by SEM, EDS, TEM, XRD and XPS. The specific surface area and thermal stability of the composite were investigated by N2 adsorption-desorption and thermogravimetric analysis, respectively. In addition, the prepared composite was used for the photocatalytic degradation of methyl orange (aqueous solution, 40 mg·L-1). It was found that the composite had a good morphology and anatase crystal structure, and Ti-O-C bond was formed between TiO2 and nanocellulose. The specific surface area of composite was increased and the thermal stability was decreased compared with the cellulose nanofiber. Moreover, the degradation rate of methyl orange was achieved as 99.72% within 30 min, and no obvious activity loss was observed after five cycles. This work might give some insights into the design of efficient photocatalysts for the treatment of organic dye wastewater.

Insights into the Structure of Rubisco from Dinoflagellates-in Silico Studies

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is one of the best studied enzymes. It is crucial for photosynthesis, and thus for all of biosphere’s productivity. There are four isoforms of this enzyme, differing by amino acid sequence composition and quaternary structure. However, there is still a group of organisms, dinoflagellates, single-cell eukaryotes, that are confirmed to possess Rubisco, but no successful purification of the enzyme of such origin, and hence a generation of a crystal structure was reported to date. Here, we are using in silico tools to generate the possible structure of Rubisco from a dinoflagellate representative, Symbiodinium sp. We selected two templates: Rubisco from Rhodospirillum rubrum and Rhodopseudomonas palustris. Both enzymes are the so-called form II Rubiscos, but the first is exclusively a homodimer, while the second one forms homo-hexamers. Obtained models show no differences in amino acids crucial for Rubisco activity. The variation was found at two closely located inserts in the C-terminal domain, of which one extends a helix and the other forms a loop. These inserts most probably do not play a direct role in the enzyme’s activity, but may be responsible for interaction with an unknown protein partner, possibly a regulator or a chaperone. Analysis of the possible oligomerization interface indicated that Symbiodinium sp. Rubisco most likely forms a trimer of homodimers, not just a homodimer. This hypothesis was empowered by calculation of binding energies. Additionally, we found that the protein of study is significantly richer in cysteine residues, which may be the cause for its activity loss shortly after cell lysis. Furthermore, we evaluated the influence of the loop insert, identified exclusively in the Symbiodinium sp. protein, on the functionality of the recombinantly expressed R. rubrum Rubisco. All these findings shed new light onto dinoflagellate Rubisco and may help in future obtainment of a native, active enzyme.