Portable Measurement System for the Characterization of Capacitive Field-Effect Sensors.

A user-friendly, portable, low-cost readout system for the on-site or point-of-care characterization of chemo- and biosensors based on an electrolyte-insulator-semiconductor capacitor (EISCAP) has been developed using a thumb-drive-sized commercial impedance analyzer. The system is controlled by a custom Python script and allows to characterize EISCAP sensors with different methods (impedance spectra, capacitance-voltage, and constant-capacitance modes), which are selected in a user interface. The performance of the portable readout system was evaluated by pH measurements and the detection of the antibiotic penicillin, hereby using EISCAPs consisting of Al/p-Si/SiO/TaO structures and compared to the results obtained with a stationary commercial impedance analyzer.

AprMH1 subtilisin from Bacillus zhangzhouensis MH1: Molecular cloning, characterization, and homology modeling.

The growing demand for robust proteases in industrial applications, particularly those based on the widely used subtilisin family, necessitates the development of novel and improved enzymes. This study reports exploration and characterization of a subtilisin, AprMH1, isolated from a recently identified Bacillus zhangzhouensis MH1 (NCBI Acct. No.

Is oxytocin related to psychiatric symptoms in adolescents with obesity?

Objectives: We aimed to investigate the relation of oxytocin receptor () gene variants ( and ) and serum oxytocin (OXT) levels with psychiatric symptoms in healthy adolescents and adolescents with obesity.

Methods: A total of 250 adolescents with obesity and 250 healthy adolescents were included in this study. Attachment properties, anxiety, and depression were evaluated with self-reports while diagnoses were ascertained with KIDDIE-SADS-PL Turkish version.

Perspectives of aminoacylases in biocatalytic synthesis of N-acyl-amino acids surfactants.

Many industrial processes are performed using harmful chemicals. The current technical synthesis of N-acyl-amino acids relies on acyl chlorides, which are typically obtained from phosgene chemistry. A greener alternative is the application of whole cells or enzymes to carry out synthesis in an environmentally friendly manner.

Development and Calibration of a Microfluidic, Chip-Based Sensor System for Monitoring the Physical Properties of Water Samples in Aquacultures.

In this work, we present a compact, bifunctional chip-based sensor setup that measures the temperature and electrical conductivity of water samples, including specimens from rivers and channels, aquaculture, and the Atlantic Ocean. For conductivity measurements, we utilize the impedance amplitude recorded via interdigitated electrode structures at a single triggering frequency. The results are well in line with data obtained using a calibrated reference instrument.

Template bacteria-free fabrication of surface imprinted polymer-based biosensor for E. coli detection using photolithographic mimics: Hacking bacterial adhesion.

As one class of molecular imprinted polymers (MIPs), surface imprinted polymer (SIP)-based biosensors show great potential in direct whole-bacteria detection. Micro-contact imprinting, that involves stamping the template bacteria immobilized on a substrate into a pre-polymerized polymer matrix, is the most straightforward and prominent method to obtain SIP-based biosensors. However, the major drawbacks of the method arise from the requirement for fresh template bacteria and often non-reproducible bacteria distribution on the stamp substrate.

Key Contributors to Signal Generation in Frequency Mixing Magnetic Detection (FMMD): An In Silico Study.

Frequency mixing magnetic detection (FMMD) is a sensitive and selective technique to detect magnetic nanoparticles (MNPs) serving as probes for binding biological targets. Its principle relies on the nonlinear magnetic relaxation dynamics of a particle ensemble interacting with a dual frequency external magnetic field. In order to increase its sensitivity, lower its limit of detection and overall improve its applicability in biosensing, matching combinations of external field parameters and internal particle properties are being sought to advance FMMD.

Actively Driven Light-Addressable Sensor/Actuator System for Automated pH Control for the Integration in Lab-On-A-Chip (LoC) Platforms.

The miniaturization of microfluidic systems usually comes at the cost of more difficult integration of sensors and actuators inside the channel. As an alternative, this work demonstrates the embedding of semiconductor-based sensor and actuator technologies that can be spatially and temporally controlled from outside the channel using light. The first element is a light-addressable potentiometric sensor, consisting of an Al/Si/SiO/SiN structure, that can measure pH changes at the SiN/electrolyte interface.

Novel recombinant aminoacylase from Paraburkholderia monticola capable of N-acyl-amino acid synthesis.

N-Acyl-amino acids can act as mild biobased surfactants, which are used, e.g., in baby shampoos.

Novel aminoacylases from Streptomyces griseus DSM 40236 and their recombinant production in Streptomyces lividans.

Amino acid-based surfactants are valuable compounds for cosmetic formulations. The chemical synthesis of acyl amino acids is conventionally performed by the Schotten-Baumann reaction using fatty acyl chlorides, but aminoacylases have also been investigated for use in biocatalytic synthesis with free fatty acids. Aminoacylases and their properties are diverse; they belong to different peptidase families and show differences in substrate specificity and biocatalytic potential.

Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System.

Immunosorbent turnip vein clearing virus (TVCV) particles displaying the IgG-binding domains D and E of protein A (PA) on every coat protein (CP) subunit (TVCV) 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 TVCV and the wild-type subgroup 3 tobamovirus.

Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822.

Subtilisins from microbial sources, especially from the Bacillaceae family, are of particular interest for biotechnological applications and serve the currently growing enzyme market as efficient and novel biocatalysts. Biotechnological applications include use in detergents, cosmetics, leather processing, wastewater treatment and pharmaceuticals. To identify a possible candidate for the enzyme market, here we cloned the gene of the subtilisin SPFA from Fictibacillus arsenicus DSM 15822 (obtained through a data mining-based search) and expressed it in Bacillus subtilis DB104.

New robust subtilisins from halotolerant and halophilic Bacillaceae.

The aim of the present study was the characterisation of three true subtilisins and one phylogenetically intermediate subtilisin from halotolerant and halophilic microorganisms. Considering the currently growing enzyme market for efficient and novel biocatalysts, data mining is a promising source for novel, as yet uncharacterised enzymes, especially from halophilic or halotolerant Bacillaceae, which offer great potential to meet industrial needs. Both halophilic bacteria Pontibacillus marinus DSM 16465 and Alkalibacillus haloalkaliphilus DSM 5271 and both halotolerant bacteria Metabacillus indicus DSM 16189 and Litchfieldia alkalitelluris DSM 16976 served as a source for the four new subtilisins SPPM, SPAH, SPMI and SPLA.

Chaperone assisted recombinant expression of a mycobacterial aminoacylase in Vibrio natriegens and Escherichia coli capable of N-lauroyl-L-amino acid synthesis.

Background: Aminoacylases are highly promising enzymes for the green synthesis of acyl-amino acids, potentially replacing the environmentally harmful Schotten-Baumann reaction. Long-chain acyl-amino acids can serve as strong surfactants and emulsifiers, with application in cosmetic industries. Heterologous expression of these enzymes, however, is often hampered, limiting their use in industrial processes.

Capacitive field-effect biosensor modified with a stacked bilayer of weak polyelectrolyte and plant virus particles as enzyme nanocarriers.

This work presents a new approach for the development of field-effect biosensors based on an electrolyte-insulator-semiconductor capacitor (EISCAP) modified with a stacked bilayer of weak polyelectrolyte and tobacco mosaic virus (TMV) particles as enzyme nanocarriers. With the aim to increase the surface density of virus particles and thus, to achieve a dense immobilization of enzymes, the negatively charged TMV particles were loaded onto the EISCAP surface modified with a positively charged poly(allylamine hydrochloride) (PAH) layer. The PAH/TMV bilayer was prepared on the TaO-gate surface by means of layer-by-layer technique.

Frequency Mixing Magnetic Detection Setup Employing Permanent Ring Magnets as a Static Offset Field Source.

Frequency mixing magnetic detection (FMMD) has been explored for its applications in fields of magnetic biosensing, multiplex detection of magnetic nanoparticles (MNP) and the determination of core size distribution of MNP samples. Such applications rely on the application of a static offset magnetic field, which is generated traditionally with an electromagnet. Such a setup requires a current source, as well as passive or active cooling strategies, which directly sets a limitation based on the portability aspect that is desired for point of care (POC) monitoring applications.

Phylogenetic survey of the subtilase family and a data-mining-based search for new subtilisins from .

The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family using a data mining approach.

A convenient ninhydrin assay in 96-well format for amino acid-releasing enzymes using an air-stable reagent.

An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP).

Synchronized, Spontaneous, and Oscillatory Detachment of Eukaryotic Cells: A New Tool for Cell Characterization and Identification.

Despite the importance of cell characterization and identification for diagnostic and therapeutic applications, developing fast and label-free methods without (bio)-chemical markers or surface-engineered receptors remains challenging. Here, we exploit the natural cellular response to mild thermal stimuli and propose a label- and receptor-free method for fast and facile cell characterization. Cell suspensions in a dedicated sensor are exposed to a temperature gradient, which stimulates synchronized and spontaneous cell-detachment with sharply defined time-patterns, a phenomenon unknown from literature.

Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101.

Halophilic and halotolerant microorganisms represent a promising source of salt-tolerant enzymes suitable for various biotechnological applications where high salt concentrations would otherwise limit enzymatic activity. Considering the current growing enzyme market and the need for more efficient and new biocatalysts, the present study aimed at the characterization of a high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101 . The protease gene was cloned and expressed in Bacillus subtilis DB104.

Field-Effect Capacitors Decorated with Ligand-Stabilized Gold Nanoparticles: Modeling and Experiments.

Nanoparticles are recognized as highly attractive tunable materials for designing field-effect biosensors with enhanced performance. In this work, we present a theoretical model for electrolyte-insulator-semiconductor capacitors (EISCAP) decorated with ligand-stabilized charged gold nanoparticles. The charged AuNPs are taken into account as additional, nanometer-sized local gates.

Electrochemical Cell-based Biosensors for Biomedical Applications.

Electrochemical cell-based biosensors have attracted increasing interest within the last 15 years, with a large number of reports generally dealing with the sensors' sensitivity, selectivity, stability, signal-to-noise ratio, spatiotemporal resolution, etc. However, only a few of them are now available as commercial products. In this review, technological advances, current challenges, and opportunities of electrochemical cell-based biosensors are presented.

Towards Multi-Analyte Detection with Field-Effect Capacitors Modified with Bioparticles as Enzyme Nanocarriers.

Utilizing an appropriate enzyme immobilization strategy is crucial for designing enzyme-based biosensors. Plant virus-like particles represent ideal nanoscaffolds for an extremely dense and precise immobilization of enzymes, due to their regular shape, high surface-to-volume ratio and high density of surface binding sites. In the present work, (TMV) particles were applied for the co-immobilization of penicillinase and urease onto the gate surface of a field-effect electrolyte-insulator-semiconductor capacitor (EISCAP) with a p-Si-SiO-TaO layer structure for the sequential detection of penicillin and urea.

An Array of On-Chip Integrated, Individually Addressable Capacitive Field-Effect Sensors with Control Gate: Design and Modelling.

The on-chip integration of multiple biochemical sensors based on field-effect electrolyte-insulator-semiconductor capacitors (EISCAP) is challenging due to technological difficulties in realization of electrically isolated EISCAPs on the same Si chip. In this work, we present a new simple design for an array of on-chip integrated, individually electrically addressable EISCAPs with an additional control gate (CG-EISCAP). The existence of the CG enables an addressable activation or deactivation of on-chip integrated individual CG-EISCAPs by simple electrical switching the CG of each sensor in various setups, and makes the new design capable for multianalyte detection without cross-talk effects between the sensors in the array.

Multiplex Detection of Magnetic Beads Using Offset Field Dependent Frequency Mixing Magnetic Detection.

Magnetic immunoassays employing Frequency Mixing Magnetic Detection (FMMD) have recently become increasingly popular for quantitative detection of various analytes. Simultaneous analysis of a sample for two or more targets is desirable in order to reduce the sample amount, save consumables, and save time. We show that different types of magnetic beads can be distinguished according to their frequency mixing response to a two-frequency magnetic excitation at different static magnetic offset fields.