In situ modulation of enzyme activity via heterogeneous catalysis utilizing solid electroplated cofactors.
ABSTRACT: During product isolation the received bioreceptors often do not exhibit a sufficient biochemical activity due to multistep dissociation and loss of cofactors. However, for bioelectrochemical applications the presence of cofactors is necessary for a successful oxidative or reductive conversion of the substrates to the products. Herein, we show how the immobilization of the required electroplated cofactors in a design of amperometric electrodes can in situ assist the activity of apo-enzymes. Compared to conventional approaches used in enzyme engineering this tailored nanoengineering methodology is superior from economic point of view, labor and time costs, storage conditions, reduced amount of waste and can fill the gap in the development of tuned bioelectrocatalysts.
Project description:To protect steel structures, zinc coatings are mostly used as a sacrificial barrier. This research aims to estimate the dissolution tendency of the electroplated and zinc-rich cold galvanized (ZRCG) coatings of a controlled thickness (35 ± 1 ?m) applied via brush and dip coating methods on the mild steel. To assess the corrosion behavior of these coated samples in 3.5% NaCl and 10% NaCl containing soil solutions, open circuit potential (OCP), cyclic polarization (CP), and electrochemical impedance spectroscopy (EIS) tests were performed. The more negative OCP and appreciably large corrosion rate of the electroplated and ZRCG coated samples in 3.5% NaCl solution highlighted the preferential dissolution of Zn coatings. However, in saline soil solution, the relatively positive OCP (>-850 mV vs. Cu/CuSO<sub>4</sub>) and lower corrosion rate of the electroplated and ZRCG coatings compared to the uncoated steel sample indicated their incapacity to protect the steel substrate. The CP scans of the zinc electroplated samples showed a positive hysteresis loop after 24 h of exposure in 3.5% NaCl and saline soil solutions attributing to the localized dissolution of the coating. Similarly, the appreciable decrease in the charge transfer resistance of the electroplated samples after 24 h of exposure corresponded to their accelerated dissolution. Compared to the localized dissolution of the electroplated and brush-coated samples, the dip-coated ZRCG samples exhibited uniform dissolution during the extended exposure (500 h) salt spray test.
Project description:In this study, we performed metal (Ag, Ni, Cu, or Pd) electroplating of core-shell metallic Ag nanowire (AgNW) networks intended for use as the anode electrode in organic light-emitting diodes (OLEDs) to modify the work function (WF) and conductivity of the AgNW networks. This low-cost and facile electroplating method enabled the precise deposition of metal onto the AgNW surface and at the nanowire (NW) junctions. AgNWs coated onto a transparent glass substrate were immersed in four different metal electroplating baths: those containing AgNO<sub>3</sub> for Ag electroplating, NiSO<sub>4</sub> for Ni electroplating, Cu<sub>2</sub>P<sub>2</sub>O<sub>7</sub> for Cu electroplating, and PdCl<sub>2</sub> for Pd electroplating. The solvated metal ions (Ag<sup>+</sup>, Ni<sup>2+</sup>, Cu<sup>2+</sup>, and Pd<sup>2+</sup>) in the respective electroplating baths were reduced to the corresponding metals on the AgNW surface in the galvanostatic mode under a constant electric current achieved by linear sweep voltammetry via an external circuit between the AgNW networks (cathode) and a Pt mesh (anode). The amount of electroplated metal was systematically controlled by varying the electroplating time. Scanning electron microscopy images showed that the four different metals (shells) were successfully electroplated on the AgNWs (core), and the nanosize-controlled electroplating process produced metal NWs with varying diameters, conductivities, optical transmittances, and WFs. The metal-electroplated AgNWs were successfully employed as the anode electrodes of the OLEDs. This facile and low-cost method of metal electroplating of AgNWs to increase their WFs and conductivities is a promising development for the fabrication of next-generation OLEDs.
Project description:Nucleoside-based cofactors are presumed to have preceded proteins. The Rossmann fold is one of the most ancient and functionally diverse protein folds, and most Rossmann enzymes utilize nucleoside-based cofactors. We analyzed an omnipresent Rossmann ribose-binding interaction: a carboxylate side chain at the tip of the second ?-strand (?2-Asp/Glu). We identified a canonical motif, defined by the ?2-topology and unique geometry. The latter relates to the interaction being bidentate (both ribose hydroxyls interacting with the carboxylate oxygens), to the angle between the carboxylate and the ribose, and to the ribose's ring configuration. We found that this canonical motif exhibits hallmarks of divergence rather than convergence. It is uniquely found in Rossmann enzymes that use different cofactors, primarily SAM (S-adenosyl methionine), NAD (nicotinamide adenine dinucleotide), and FAD (flavin adenine dinucleotide). Ribose-carboxylate bidentate interactions in other folds are not only rare but also have a different topology and geometry. We further show that the canonical geometry is not dictated by a physical constraint--geometries found in noncanonical interactions have similar calculated bond energies. Overall, these data indicate the divergence of several major Rossmann-fold enzyme classes, with different cofactors and catalytic chemistries, from a common pre-LUCA (last universal common ancestor) ancestor that possessed the ?2-Asp/Glu motif.
Project description:Recent investigations into cardiac or nervous tissues call for systems that are able to electrically record in 3D as opposed to 2D. Typically, challenging microfabrication steps are required to produce 3D microelectrode arrays capable of recording at the desired position within the tissue of interest. As an alternative, additive manufacturing is becoming a versatile platform for rapidly prototyping novel sensors with flexible geometric design. In this work, 3D MEAs for cell-culture applications were fabricated using a piezoelectric inkjet printer. The aspect ratio and height of the printed 3D electrodes were user-defined by adjusting the number of deposited droplets of silver nanoparticle ink along with a continuous printing method and an appropriate drop-to-drop delay. The Ag 3D MEAs were later electroplated with Au and Pt in order to reduce leakage of potentially cytotoxic silver ions into the cellular medium. The functionality of the array was confirmed using impedance spectroscopy, cyclic voltammetry, and recordings of extracellular potentials from cardiomyocyte-like HL-1 cells.
Project description:«Crude» extracts obtained via simple ultrasonic disintegration of microbial cell membrane are perspective bioelectrocatalysts. This extract contains all the necessary enzymes and cofactors required for oxidative or reductive conversion. The technology of synthesis of «crude extract» is simpler and less costly in comparison with technology of obtaining pure enzymes. Dialysis of the obtained extracts was performed with different molecular weight cut-off (3.5 kDa, 12-14 kDa, 25 kDa, 50 kDa). The obtained data show that after dialysis extracts lose their dehydrogenase and bioelectrocatalytic activity due to the loss of cofactors. However, the addition of NAD and NADP cofactors leads to a recovery of activity. The obtained data demonstrate that the concentration of the cofactor directly affects the rate of the bioelectrocatalytic reaction. Also, the obtained data indicate that the composition of the enzyme systems of the extract includes succinate dehydrogenase. Analyzing this data set can provide insight on increase of the electrocatalytic activity of a new type of bioelectrocatalyst.
Project description:This communication presents fluidic self-assembly of Si-chip on a sequentially electroplated multilayer solder bump with tailored transformation imprinted melting points. The multilayer solder bump is a lead free ternary solder system, which provides a route to transform the melting point of interconnects for applications in solder directed fluidic self-assembly. The outermost metal layers form a low melting point Bi33.7In66.3 solder shell (72?°C). This solder shell enables fluidic self-assembly and self-alignment of freely in water suspended Si-dies at relatively low temperature (75?°C) leading to well-ordered chip arrays. The reduction of the free surface energy of the shell-water interface provides the driving force for the self-assembly. The lowermost metal layer is a high melting point solder and acts as a core. After the self-assembly is complete, a short reflow causes the transformation of the core and the shell yielding a stable high melting point solder with adjustable melting points. The chosen ternary solder system enables the realization of interconnects with melting points in the range of 112?°C to 206?°C.
Project description:Critical drivers of cancer progression are likely controlled through the actions of transcription factors and cofactors that bind to the genome and form enhancers that stimulate gene expression. We present ChIP-seq analysis of key transcriptional regulators, cofactors and histone modifications that indicate transcriptional activity across a range of different cancer cells. ChIP-seq was performed against the cofactors Med1, Brd4 and CDK7 in MM1.S multiple myeloma cells; Brd4 in Sk-MEL-5 melanoma cells; and H3K9/K14 acetylation in MV4;11 leukemia cells,
Project description:The recently identified glmS ribozyme revealed that RNA enzymes, like protein enzymes, are capable of using small molecules as catalytic cofactors to promote chemical reactions. Flavin mononucleotide (FMN), S-adenosyl methionine (SAM), adenosyl cobalamin (AdoCbl), and thiamine pyrophosphate (TPP) are known ligands for RNA riboswitches in the control of gene expression, but are also catalytically powerful and ubiquitous cofactors in protein enzymes. If RNA, instead of just binding these molecules, could harness the chemical potential of the cofactor, it would significantly expand the enzymatic repertoire of ribozymes. Here we review the chemistry of AdoCbl, SAM, FMN, and TPP in protein enzymology and speculate on how these cofactors might have been used by ribozymes in the prebiotic RNA World or may still find application in modern biology.
Project description:<h4>Motivation</h4>Organic enzyme cofactors are involved in many enzyme reactions. Therefore, the analysis of cofactors is crucial to gain a better understanding of enzyme catalysis. To aid this, we have created the CoFactor database.<h4>Results</h4>CoFactor provides a web interface to access hand-curated data extracted from the literature on organic enzyme cofactors in biocatalysis, as well as automatically collected information. CoFactor includes information on the conformational and solvent accessibility variation of the enzyme-bound cofactors, as well as mechanistic and structural information about the hosting enzymes.<h4>Availability</h4>The database is publicly available and can be accessed at http://www.ebi.ac.uk/thornton-srv/databases/CoFactor.
Project description:Synthetic nicotinamide cofactors are analogues of the natural cofactors used by oxidoreductases as redox intermediates. Their ability to be fine-tuned makes these biomimetics an attractive alternative to the natural cofactors in terms of stability, reactivity, and cost. The following mini-review focuses on the current state of the art of those biomimetics in enzymatic processes.