Refine
Year of publication
Document Type
- Article (501)
- Conference Proceeding (39)
- Part of a Book (8)
- Doctoral Thesis (3)
- Book (2)
- Other (2)
Language
- English (555) (remove)
Keywords
- Biosensor (6)
- biosensors (4)
- frequency mixing magnetic detection (4)
- LAPS (3)
- Label-free detection (3)
- Light-addressable potentiometric sensor (3)
- capacitive field-effect sensor (3)
- field-effect sensor (3)
- hydrogen peroxide (3)
- magnetic nanoparticles (3)
- tobacco mosaic virus (TMV) (3)
- Acyl-amino acids (2)
- Aminoacylase (2)
- Bacillaceae (2)
- Bacillus atrophaeus (2)
- Biotechnological application (2)
- Calorimetric gas sensor (2)
- Capacitive field-effect sensor (2)
- Hydrogen peroxide (2)
- Raman spectroscopy (2)
- Subtilases (2)
- Subtilisin (2)
- Tobacco mosaic virus (TMV) (2)
- acetoin (2)
- capacitive field-effect sensors (2)
- gold nanoparticles (2)
- light-addressable potentiometric sensor (2)
- penicillinase (2)
- sterilisation (2)
- (Bio)degradation (1)
- Acylation (1)
- Alginate beads (1)
- Alkalihalobacillus okhensis (1)
- Bacillus atrophaeus spores (1)
- Bio-Sensors (1)
- Bioabsorbable (1)
- Biocatalysis (1)
- Biosensorik (1)
- Biosurfactants (1)
- Broad pH spectrum (1)
- CNOT (1)
- CRISPR/Cas9 (1)
- Capacitive field-effect (1)
- Capacitive model (1)
- Chaperone (1)
- Chaperone co-expression (1)
- Chemical images (1)
- Chemical imaging (1)
- Chemical imaging sensor (1)
- Chemical sensor (1)
- Coat protein (1)
- C–V method (1)
- DNA biosensor (1)
- DPA (dipicolinic acid) (1)
- Dehydrogenase (1)
- Detergent protease (1)
- Diaphorase (1)
- EIS capacitive sensor (1)
- Electrolyte–insulator–semiconductor (1)
- Enzymatic biosensor (1)
- Enzyme coverage (1)
- Enzyme logic gate (1)
- Enzyme nanocarrier (1)
- Extracellular enzymes (1)
- Field effect (1)
- Field-effect biosensor (1)
- Field-effect device (1)
- Field-effect sensor (1)
- GaAs hot electron injector (1)
- Gas sensor (1)
- Glucose biosensor (1)
- Glucose oxidase (1)
- Gold nanoparticles (1)
- Gunn diode (1)
- Halotolerant protease (1)
- Heavy metal detection (1)
- Hypersecretion (1)
- ISFET (1)
- Impedance spectroscopy (1)
- Inclusion bodies (1)
- Lab-on-Chip (1)
- Layer-by-layer adsorption (1)
- LbL films (1)
- MOS (1)
- Marker-free mutagenesis (1)
- Multi-sensor system (1)
- Multianalyte detection (1)
- Multicell (1)
- Multiplexing (1)
- Nano Materials (1)
- Nanomaterial (1)
- Nanostructuring (1)
- Nanotechnologie (1)
- Negative impedance convertor (1)
- O2 plasma (1)
- Organic light-emitting diode display (1)
- Penicillin (1)
- Plant virus (1)
- Poly(allylamine hydrochloride) (1)
- Poly(d,l-lacticacid) (1)
- Polyimide (1)
- Polylactide acid (1)
- Potentiometry (1)
- Real-time monitoring (1)
- Resistive temperature detector (1)
- Resonance-mode measurement (1)
- Silk fibroin (1)
- Simultaneous determination (1)
- Sn₃O₄ (1)
- Stenotrophomonas maltophilia (1)
- Sterilisation process (1)
- Streptomyces griseus (1)
- Streptomyces lividans (1)
- TMV adsorption (1)
- Ta₂O₅ gate (1)
- Tobacco mosaic virus (1)
- Uracil-phosphoribosyltransferase (1)
- Vibrio natriegens (1)
- Wafer (1)
- XOR (1)
- Zeta potential (1)
- acetoin reductase (1)
- actuator-sensor system (1)
- alcoholic beverages (1)
- aminooctanethiol (1)
- amperometric biosensors (1)
- annealing (1)
- artificial olfactory image (1)
- aseptic parameters (1)
- aspergillus (1)
- atomic layer deposition (1)
- bi-enzyme biosensor (1)
- bioburdens (1)
- biocompatible (1)
- biocompatible materials (1)
- biodegradabl (1)
- biodegradable electronic devices (1)
- biosensor (1)
- calorimetric gas sensor (1)
- calorimetric gas sensor;hydrogen peroxide;wireless sensor system (1)
- capacitive EIS sensor (1)
- capacitive electrolyte–insulator–semiconductor sensors (1)
- capacitive field-effect biosensor (1)
- capacitive model (1)
- capillary micro-droplet cell (1)
- carbon electrodes (1)
- catalytic decomposition (1)
- catalytic metal (1)
- chemical sensor (1)
- colorization (1)
- control gate (1)
- coupled Néel–Brownian relaxation dynamics (1)
- detection of charged macromolecules (1)
- detergent protease (1)
- electrolyte-insulator semiconductor sensor (EIS) (1)
- electrolyte-insulator-semiconductor capacitors (1)
- electronic nose (1)
- encapsulation materials (1)
- endospores (1)
- enzymatic biosensor (1)
- enzyme cascade (1)
- enzyme immobilization (1)
- enzyme kinetics (1)
- enzyme-logic gate (1)
- equivalent circuit (1)
- fibroin (1)
- field-effect structure (1)
- filamentous fungi (1)
- frequency mixing (1)
- gas sensor (1)
- gaseous hydrogen peroxide (1)
- genome engineering (1)
- glucose (1)
- glucose oxidase (GOx) (1)
- graphene oxide (1)
- halotolerant protease (1)
- heavy metals (1)
- high-alkaline subtilisin (1)
- horseradish peroxidase (HRP) (1)
- hydroxylation (1)
- immobilization (1)
- key performance indicators (1)
- layer expansion (1)
- layer-by-layer technique (1)
- light-addressable electrode (1)
- light-addressing technologies (1)
- magnetic actuation (1)
- magnetic beads (1)
- magnetic biosensing (1)
- magnetic relaxation (1)
- magnetic sandwich immunoassay (1)
- magnetic sensing (1)
- magnetic sensors (1)
- magnetic separation (1)
- magnetic tweezers (1)
- magnetophoretic velocity (1)
- metal-oxide-semiconductor structure (1)
- microfluidics (1)
- micromagnetic simulation (1)
- microwave generation (1)
- multi-sensing platform (1)
- multianalyte detection (1)
- multiparametric immunoassays (1)
- multiplex detection (1)
- nanobelts (1)
- nanomaterials (1)
- nanoparticle coverage (1)
- on-chip integrated addressable EISCAP sensors (1)
- optical sensor setup (1)
- optical spore trapping (1)
- optical trapping (1)
- organic PVC membranes (1)
- organosilanes (1)
- oxidative stable protease (1)
- pH sensors (1)
- pattern-size reduction (1)
- penicillin (1)
- photoelectrochemistry (1)
- plant virus detection (1)
- plug-based microfluidic device (1)
- polyaniline (1)
- polystyrene sulfonate (1)
- recombinant expression (1)
- scanned light pulse technique (1)
- self-aligned patterning (1)
- silanization (1)
- spore kill rate (1)
- sterility (1)
- sterility tests (1)
- sterilization (1)
- sterilization conditions (1)
- sterilization efficacy (1)
- sterilization methods (1)
- superparamagnetic bead (1)
- superparamagnetic nanoparticles (1)
- surface functionalization (1)
- temperature (1)
- thin-film microsensors (1)
- titanium dioxide photoanode (1)
- turnip vein clearing virus (TVCV) (1)
- ultrathin gate insulators (1)
- urease (1)
- validation methods (1)
- visualization (1)
- wafer-level testing (1)
- α-aminoacylase (1)
- ε-lysine acylase (1)
Institute
- INB - Institut für Nano- und Biotechnologien (555) (remove)
Plant virus-like particles, and in particular, tobacco mosaic virus (TMV) particles, are increasingly being used in nano- and biotechnology as well as for biochemical sensing purposes as nanoscaffolds for the high-density immobilization of receptor molecules. The sensitive parameters of TMV-assisted biosensors depend, among others, on the density of adsorbed TMV particles on the sensor surface, which is affected by both the adsorption conditions and surface properties of the sensor. In this work, Ta₂O₅-gate field-effect capacitive sensors have been applied for the label-free electrical detection of TMV adsorption. The impact of the TMV concentration on both the sensor signal and the density of TMV particles adsorbed onto the Ta₂O₅-gate surface has been studied systematically by means of field-effect and scanning electron microscopy methods. In addition, the surface density of TMV particles loaded under different incubation times has been investigated. Finally, the field-effect sensor also demonstrates the label-free detection of penicillinase immobilization as model bioreceptor on TMV particles.
Electrolyte-insulator-semiconductor (EIS) field-effect sensors belong to a new generation of electronic chips for biochemical sensing, enabling a direct electronic readout. The review gives an overview on recent advances and current trends in the research and development of chemical sensors and biosensors based on the capacitive field-effect EIS structure—the simplest field-effect device, which represents a biochemically sensitive capacitor. Fundamental concepts, physicochemical phenomena underlying the transduction mechanism and application of capacitive EIS sensors for the detection of pH, ion concentrations, and enzymatic reactions, as well as the label-free detection of charged molecules (nucleic acids, proteins, and polyelectrolytes) and nanoparticles, are presented and discussed.
An acetoin biosensor based on a capacitive electrolyte–insulator–semiconductor (EIS) structure modified with the enzyme acetoin reductase, also known as butane-2,3-diol dehydrogenase (Bacillus clausii DSM 8716ᵀ), is applied for acetoin detection in beer, red wine, and fermentation broth samples for the first time. The EIS sensor consists of an Al/p-Si/SiO₂/Ta₂O₅ layer structure with immobilized acetoin reductase on top of the Ta₂O₅ transducer layer by means of crosslinking via glutaraldehyde. The unmodified and enzyme-modified sensors are electrochemically characterized by means of leakage current, capacitance–voltage, and constant capacitance methods, respectively.
Electrolyte-insulator-semiconductor capacitors (EISCAP) belong to field-effect sensors having an attractive transducer architecture for constructing various biochemical sensors. In this study, a capacitive model of enzyme-modified EISCAPs has been developed and the impact of the surface coverage of immobilized enzymes on its capacitance-voltage and constant-capacitance characteristics was studied theoretically and experimentally. The used multicell arrangement enables a multiplexed electrochemical characterization of up to sixteen EISCAPs. Different enzyme coverages have been achieved by means of parallel electrical connection of bare and enzyme-covered single EISCAPs in diverse combinations. As predicted by the model, with increasing the enzyme coverage, both the shift of capacitance-voltage curves and the amplitude of the constant-capacitance signal increase, resulting in an enhancement of analyte sensitivity of the EISCAP biosensor. In addition, the capability of the multicell arrangement with multi-enzyme covered EISCAPs for sequentially detecting multianalytes (penicillin and urea) utilizing the enzymes penicillinase and urease has been experimentally demonstrated and discussed.
Algal polysaccharides (extracellular polysaccharides) and carbon nanotubes (CNTs) were adsorbed on dioctadecyldimethylammonium bromide Langmuir monolayers to serve as a matrix for the incorporation of urease. The physicochemical properties of the supramolecular system as a monolayer at the air–water interface were investigated by surface pressure–area isotherms, surface potential–area isotherms, interfacial shear rheology, vibrational spectroscopy, and Brewster angle microscopy. The floating monolayers were transferred to hydrophilic solid supports, quartz, mica, or capacitive electrolyte–insulator–semiconductor (EIS) devices, through the Langmuir–Blodgett (LB) technique, forming mixed films, which were investigated by quartz crystal microbalance, fluorescence spectroscopy, and field emission gun scanning electron microscopy. The enzyme activity was studied with UV–vis spectroscopy, and the feasibility of the thin film as a urea sensor was essayed in an EIS sensor device. The presence of CNT in the enzyme–lipid LB film not only tuned the catalytic activity of urease but also helped to conserve its enzyme activity. Viability as a urease sensor was demonstrated with capacitance–voltage and constant capacitance measurements, exhibiting regular and distinctive output signals over all concentrations used in this work. These results are related to the synergism between the compounds on the active layer, leading to a surface morphology that allowed fast analyte diffusion owing to an adequate molecular accommodation, which also preserved the urease activity. This work demonstrates the feasibility of employing LB films composed of lipids, CNT, algal polysaccharides, and enzymes as EIS devices for biosensing applications.
In this paper, carbon nanotubes (CNTs) were incorporated in penicillinase-phospholipid Langmuir and Langmuir–Blodgett (LB) films to enhance the enzyme catalytic properties. Adsorption of the penicillinase and CNTs at dimyristoylphosphatidic acid (DMPA) monolayers at the air–water interface was investigated by surface pressure–area isotherms, vibrational spectroscopy, and Brewster angle microscopy. The floating monolayers were transferred to solid supports through the LB technique, forming mixed DMPA-CNTs-PEN films, which were investigated by quartz crystal microbalance, vibrational spectroscopy, and atomic force microscopy. Enzyme activity was studied with UV–vis spectroscopy and the feasibility of the supramolecular device nanostructured as ultrathin films were essayed in a capacitive electrolyte–insulator–semiconductor (EIS) sensor device. The presence of CNTs in the enzyme–lipid LB film not only tuned the catalytic activity of penicillinase but also helped conserve its enzyme activity after weeks, showing increased values of activity. Viability as penicillin sensor was demonstrated with capacitance/voltage and constant capacitance measurements, exhibiting regular and distinctive output signals over all concentrations used in this work. These results may be related not only to the nanostructured system provided by the film, but also to the synergism between the compounds on the active layer, leading to a surface morphology that allowed a fast analyte diffusion because of an adequate molecular accommodation, which also preserved the penicillinase activity. This work therefore demonstrates the feasibility of employing LB films composed of lipids, CNTs, and enzymes as EIS devices for biosensing applications.
Members of the species Bacillus pumilus get more and more in focus of the biotechnological industry as potential new production strains. Based on exoproteome analysis, B. pumilus strain Jo2, possessing a high secretion capability, was chosen for an omics-based investigation. The proteome and metabolome of B. pumilus cells growing either in minimal or complex medium was analyzed. In total, 1542 proteins were identified in growing B. pumilus cells, among them 1182 cytosolic proteins, 297 membrane and lipoproteins and 63 secreted proteins. This accounts for about 43% of the 3616 proteins encoded in the B. pumilus Jo2 genome sequence. By using GC–MS, IP-LC/MS and H NMR methods numerous metabolites were analyzed and assigned to reconstructed metabolic pathways. In the genome sequence a functional secretion system including the components of the Sec- and Tat-secretion machinery was found. Analysis of the exoproteome revealed secretion of about 70 proteins with predicted secretion signals. In addition, selected production-relevant genome features such as restriction modification systems and NRPS clusters of B. pumilus Jo2 are discussed.
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.
Results
We identified a novel mycobacterial aminoacylase gene from Mycolicibacterium smegmatis MKD 8, cloned and expressed it in Escherichia coli and Vibrio natriegens using the T7 overexpression system. The recombinant enzyme was prone to aggregate as inclusion bodies, and while V. natriegens Vmax™ could produce soluble aminoacylase upon induction with isopropyl β-d-1-thiogalactopyranoside (IPTG), E. coli BL21 (DE3) needed autoinduction with lactose to produce soluble recombinant protein. We successfully conducted a chaperone co-expression study in both organisms to further enhance aminoacylase production and found that overexpression of chaperones GroEL/S enhanced aminoacylase activity in the cell-free extract 1.8-fold in V. natriegens and E. coli. Eventually, E. coli ArcticExpress™ (DE3), which co-expresses cold-adapted chaperonins Cpn60/10 from Oleispira antarctica, cultivated at 12 °C, rendered the most suitable expression system for this aminoacylase and exhibited twice the aminoacylase activity in the cell-free extract compared to E. coli BL21 (DE3) with GroEL/S co-expression at 20 °C. The purified aminoacylase was characterized based on hydrolytic activities, being most stable and active at pH 7.0, with a maximum activity at 70 °C, and stability at 40 °C and pH 7.0 for 5 days. The aminoacylase strongly prefers short-chain acyl-amino acids with smaller, hydrophobic amino acid residues. Several long-chain amino acids were fairly accepted in hydrolysis as well, especially N-lauroyl-L-methionine. To initially evaluate the relevance of this aminoacylase for the synthesis of N-acyl-amino acids, we demonstrated that lauroyl-methionine can be synthesized from lauric acid and methionine in an aqueous system.
Conclusion
Our results suggest that the recombinant enzyme is well suited for synthesis reactions and will thus be further investigated.