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A novel peptidyl-lys metalloendopeptidase (Tc-LysN) from Tramates coccinea was recombinantly expressed in Komagataella phaffii using the native pro-protein sequence. The peptidase was secreted into the culture broth as zymogen (~38 kDa) and mature enzyme (~19.8 kDa) simultaneously. The mature Tc-LysN was purified to homogeneity with a single step anion-exchange chromatography at pH 7.2. N-terminal sequencing using TMTpro Zero and mass spectrometry of the mature Tc-LysN indicated that the pro-peptide was cleaved between the amino acid positions 184 and 185 at the Kex2 cleavage site present in the native pro-protein sequence. The pH optimum of Tc-LysN was determined to be 5.0 while it maintained ≥60% activity between pH values 4.5—7.5 and ≥30% activity between pH values 8.5—10.0, indicating its broad applicability. The temperature maximum of Tc-LysN was determined to be 60 °C. After 18 h of incubation at 80 °C, Tc-LysN still retained ~20% activity. Organic solvents such as methanol and acetonitrile, at concentrations as high as 40% (v/v), were found to enhance Tc-LysN’s activity up to ~100% and ~50%, respectively. Tc-LysN’s thermostability, ability to withstand up to 8 M urea, tolerance to high concentrations of organic solvents, and an acidic pH optimum make it a viable candidate to be employed in proteomics workflows in which alkaline conditions might pose a challenge. The nano-LC-MS/MS analysis revealed bovine serum albumin (BSA)’s sequence coverage of 84% using Tc-LysN which was comparable to the sequence coverage of 90% by trypsin peptides.
Photovoltaic-heat pump (PV-HP) combinations with battery and energy management systems are becoming increasingly popular due to their ability to increase the autarchy and utilization of self-generated PV electricity. This trend is driven by the ongoing electrification of the heating sector and the growing disparity between growing electricity costs and reducing feed-in tariffs in Germany. Smart control strategies can be employed to control and optimize the heat pump operation to achieve higher self-consumption of PV electricity. This work presents the evaluation results of a smart-grid ready controlled PV-HP-battery system in a single-family household in Germany, using 1-minute-high-resolution field measurement data. Within 12 months evaluation period, a self-consumption of 43% was determined. The solar fraction of the HP amounts to 36%, enabled also due to higher set temperatures for space heating and domestic hot water production. Accordingly, the SPF decreases by 4.0% the space heating and by 5.7% in the domestic hot water mode. The combined seasonal performance factor for the heat pump system increases from 4.2 to 6.7, when only considering the electricity taken from the grid and disregarding the locally generated electricity supplied from photovoltaic and battery units.
Batteries typically consist of multiple individual cells connected in series. Here we demonstrate single-cell state of charge (SOC) and state of health (SOH) diagnosis in a 24 V class lithium-ion battery. To this goal, we introduce and apply a novel, highly efficient algorithm based on a voltage-controlled model (VCM). The battery, consisting of eight single cells, is cycled over a duration of five months under a simple cycling protocol between 20 % and 100 % SOC. The cell-to-cell standard deviations obtained with the novel algorithm were 1.25 SOC-% and 1.07 SOH-% at beginning of cycling. A cell-averaged capacity loss of 9.9 % after five months cycling was observed. While the accuracy of single-cell SOC estimation was limited (probably owed to the flat voltage characteristics of the lithium iron phosphate, LFP, chemistry investigated here), single-cell SOH estimation showed a high accuracy (2.09 SOH-% mean absolute error compared to laboratory reference tests). Because the algorithm does not require observers, filters, or neural networks, it is computationally very efficient (three seconds analysis time for the complete data set consisting of eight cells with approx. 780.000 measurement points per cell).
In a randomized controlled cross-over study ten male runners (26.7 ± 4.9 years; recent 5-km time: 18:37 ± 1:07 min:s) performed an incremental treadmill test (ITT) and a 3-km time trial (3-km TT) on a treadmill while wearing either carbon fiber insoles with downwards curvature or insoles made of butyl rubber (control condition) in light road racing shoes (Saucony Fastwitch 9). Oxygen uptake, respiratory exchange ratio, heart rate, blood lactate concentration, stride frequency, stride length and time to exhaustion were assessed during ITT. After ITT, all runners rated their perceived exertion, perceived shoe comfort and perceived shoe performance. Running time, heart rate, blood lactate levels, stride frequency and stride length were recorded during, and shoe comfort and shoe performance after, the 3-km TT. All parameters obtained during or after the ITT did not differ between the two conditions [range: p = 0.188 to 0.948 (alpha value: 0.05); Cohen's d = 0.021 to 0.479] despite the rating of shoe comfort showing better scores for the control insoles (p = 0.001; d = −1.646). All parameters during and after the 3-km TT showed no differences (p = 0.200 to 1.000; d = 0.000 to 0.501) between both conditions except for shoe comfort showing better scores for control insoles (p = 0.017; d = −0.919). Running with carbon fiber insoles with downwards curvature did not change running performance or any submaximal or maximal physiological or biomechanical parameter and perceived exertion compared to control condition. Shoe comfort is impaired while running with carbon fiber insoles. Wearing carbon fiber insoles with downwards curvature during treadmill running is not beneficial when compared to running with control insoles.
Injury prevention is essential in running due to the risk of overuse injury development. Tailoring running shoes to individual needs may be a promising strategy to reduce this risk. Novel manufacturing processes allow the production of individualised running shoes that incorporate features that meet individual biomechanical and experiential needs. However, specific ways to individualise footwear to reduce injury risk are poorly understood. Therefore, this scoping review provides an overview of (1) footwear design features that have the potential for individualisation; and (2) the literature on the differential responses to footwear design features between selected groups of individuals. These purposes focus exclusively on reducing the risk of overuse injuries. We included studies in the English language on adults that analysed: (1) potential interaction effects between footwear design features and subgroups of runners or covariates (e.g., age, sex) for running-related biomechanical risk factors or injury incidences; (2) footwear comfort perception for a systematically modified footwear design feature. Most of the included articles (n = 107) analysed male runners. Female runners may be more susceptible to footwear-induced changes and overuse injury development; future research should target more heterogonous sampling. Several footwear design features (e.g., midsole characteristics, upper, outsole profile) show potential for individualisation. However, the literature addressing individualised footwear solutions and the potential to reduce biomechanical risk factors is limited. Future studies should leverage more extensive data collections considering relevant covariates and subgroups while systematically modifying isolated footwear design features to inform footwear individualisation.
For the treatment of bone defects, biodegradable, compressive biomaterials are needed as replacements that degrade as the bone regenerates. The problem with existing materials has either been their insufficient mechanical strength or the excessive differences in their elastic modulus, leading to stress shielding and eventual failure. In this study, the compressive strength of CPC ceramics (with a layer thickness of more than 12 layers) was compared with sintered β-TCP ceramics. It was assumed that as the number of layers increased, the mechanical strength of 3D-printed scaffolds would increase toward the value of sintered ceramics. In addition, the influence of the needle inner diameter on the mechanical strength was investigated. Circular scaffolds with 20, 25, 30, and 45 layers were 3D printed using a 3D bioplotter, solidified in a water-saturated atmosphere for 3 days, and then tested for compressive strength together with a β-TCP sintered ceramic using a Zwick universal testing machine. The 3D-printed scaffolds had a compressive strength of 41.56 ± 7.12 MPa, which was significantly higher than that of the sintered ceramic (24.16 ± 4.44 MPa). The 3D-printed scaffolds with round geometry reached or exceeded the upper limit of the compressive strength of cancellous bone toward substantia compacta. In addition, CPC scaffolds exhibited more bone-like compressibility than the comparable β-TCP sintered ceramic, demonstrating that the mechanical properties of CPC scaffolds are more similar to bone than sintered β-TCP ceramics.
Recently, photovoltaic (PV) with energy storage systems (ESS) have been widely adopted in buildings to overcome growing power demands and earn financial benefits. The overall energy cost can be optimized by combining a well-sized hybrid PV/ESS system with an efficient energy management system (EMS). Generally, EMS is implemented within the overall functions of the Building Automation System (BAS). However, due to its limited computing resources, BAS cannot handle complex algorithms that aim to optimize energy use in real-time under different operating conditions. Furthermore, islanding the building's local network to maximize the PV energy share represents a challenging task due to the potential technical risks. In this context, this article addresses an improved approach based on upgrading the BAS data analytics capability by means of an edge computing technology. The edge communicates with the BAS low-level controller using a serial communication protocol. Taking advantage of the high computing ability of the edge device, an optimization-based EMS of the PV/ESS hybrid system is implemented. Different testing scenarios have been carried out on a real prototype with different weather conditions, and the results show the implementation feasibility and technical performance of such advanced EMS for the management of building energy resources. It has also been proven to be feasible and advantageous to operate the local energy network in island mode while ensuring system safety. Additionally, an estimated energy saving improvement of 6.23 % has been achieved using optimization-based EMS compared to the classical rule-based EMS, with better ESS constraints fulfillment.
Enzyme‐assisted HPTLC method for the simultaneous analysis of inositol phosphates and phosphate
(2023)
Background
The analysis of myo‐inositol phosphates (InsPx) released by phytases during phytic acid degradation is challenging and time‐consuming, particularly in terms of sample preparation, isomer separation, and detection. However, a fast and robust analysis method is crucial when screening for phytases during protein engineering approaches, which result in a large number of samples, to ensure reliable identification of promising novel enzymes or target variants with improved characteristics, for example, pH range, thermal stability, and phosphate release kinetics.
Results
The simultaneous analysis of several InsPx (InsP1‐InsP4 and InsP5 + 6) as well as free phosphate was established on cellulose HPTLC plates using a buffered mobile phase. Inositol phosphates were subsequently stained using a novel enzyme‐assisted staining procedure. Immobilized InsPx were hydrolyzed by a phytase solution of Quantum® Blueliquid 5G followed by a molybdate reagent derivatization. Resulting blue zones were captured by DAD scan. The method shows good repeatability (intra‐day and intra‐lab) with maximum deviations of the Rf value of 0.01. The HPTLC method was applied to three commercially available phytases at two pH levels relevant to the gastrointestinal tract of poultry (pH 5.5 and pH 3.6) to observe their phytate degradation pattern and thus visualize their InsPx fingerprint.
Conclusion
This HPTLC method presents a semi‐high‐throughput analysis for the simultaneous analysis of phytic acid and the resulting lower inositol phosphates after its enzymatic hydrolysis and is also an effective tool to visualize the InsPx fingerprints and possible accumulations of inositol phosphates.
Building energy management systems (BEMSs), dedicated to sustainable buildings, may have additional duties, such as hosting efficient energy management systems (EMSs) algorithms. This duty can become crucial when operating renewable energy sources (RES) and eventual electric energy storage systems (ESSs). Sophisticated EMS approaches that aim to manage RES and ESSs in real time may need high computing capabilities that BEMSs typically cannot provide. This article addresses and validates a fuzzy logic-based EMS for the optimal management of photovoltaic (PV) systems with lead-acid ESSs using an edge computing technology. The proposed method is tested on a real smart grid prototype in comparison with a classical rule-based EMS for different weather conditions. The goal is to investigate the efficacy of islanding the building local network as a control command, along with ESS power control. The results show the implementation feasibility and performance of the fuzzy algorithm in the optimal management of ESSs in both operation modes: grid-connected and islanded modes.
Immunosorbent turnip vein clearing virus (TVCV) particles displaying the IgG-binding domains D and E of Staphylococcus aureus protein A (PA) on every coat protein (CP) subunit (TVCVPA) were purified from plants via optimized and new protocols. The latter used polyethylene glycol (PEG) raw precipitates, from which virions were selectively re-solubilized in reverse PEG concentration gradients. This procedure improved the integrity of both TVCVPA and the wild-type subgroup 3 tobamovirus. TVCVPA could be loaded with more than 500 IgGs per virion, which mediated the immunocapture of fluorescent dyes, GFP, and active enzymes. Bi-enzyme ensembles of cooperating glucose oxidase and horseradish peroxidase were tethered together on the TVCVPA carriers via a single antibody type, with one enzyme conjugated chemically to its Fc region, and the other one bound as a target, yielding synthetic multi-enzyme complexes. In microtiter plates, the TVCVPA-displayed sugar-sensing system possessed a considerably increased reusability upon repeated testing, compared to the IgG-bound enzyme pair in the absence of the virus. A high coverage of the viral adapters was also achieved on Ta2O5 sensor chip surfaces coated with a polyelectrolyte interlayer, as a prerequisite for durable TVCVPA-assisted electrochemical biosensing via modularly IgG-assembled sensor enzymes.