Neuromorphic Computing-Assisted Triboelectric Capacitive-Coupled Tactile Sensor Array for Wireless Mixed Reality Interaction.

Flexible tactile sensors show promise for artificial intelligence applications due to their biological adaptability and rapid signal perception. Triboelectric sensors enable active dynamic tactile sensing, while integrating static pressure sensing and real-time multichannel signal transmission is key for further development. Here, we propose an integrated structure combining a capacitive sensor for static spatiotemporal mapping and a triboelectric sensor for dynamic tactile recognition. A liquid metal-based flexible dual-mode triboelectric-capacitive-coupled tactile sensor (TCTS) array of 4 × 4 pixels achieves a spatial resolution of 7 mm, exhibiting a pressure detection limit of 0.8 Pa and a fast response of 6 ms. Furthermore, neuromorphic computing using the MXene-based synaptic transistor achieves 100% recognition accuracy of handwritten numbers/letters within 90 epochs based on dynamic triboelectric signals collected by the TCTS array, and cross-spatial information communication from the perceived multichannel tactile data is realized in the mixed reality space. The results illuminate considerable application possibilities of dual-mode tactile sensing technology in human-machine interfaces and advanced robotics.

Surface Morphology Analysis of Knit Structure-Based Triboelectric Nanogenerator for Enhancing the Transfer Charge.

Harvesting waste biomechanical energy has provided a promising approach to improve the power supplement of wearable devices for extending usage life. Surface morphology is a significant factor for enhancing output performance of triboelectric nanogenerator; however, there is a limitation for evaluating the morphology of the surface and its impact on power generation. To evaluate the relationship between the surface morphology and transfer charge, there is a mathematical theory that is the fractal geometry theory that has been proposed to analyze the characteristic of irregular surface morphology. This theory provided a good understanding of the contact area and roughness of the surface. We have designed three categories of knit structures with cord appearance by using a flat knitting machine and analyzed their surface characteristics. Meanwhile, the geometric structures can be demonstrated through the fractal dimension for evaluating the generated output performance during contacting and separation. The present research exhibits that, with the increasing number of knitted units, the triboelectric power-generation performance continued to reduce due to the available contact area decreasing. After calculating the fractal dimension of different knit structures, the m*n rib structures show the high transfer charge when the fractal dimension is close to number one, especially the fractal dimension of the 1*1 rib structure that can reach 0.99. The fractal theory can be further used as an approach to evaluate the influence on the output performance of irregular surface morphology, unrelated to the uniform convex unit distraction. The result of this research also demonstrated the feasibility of a knitted-based triboelectric nanogenerator in scavenging biomechanical energy for powering portable electronics integrated into garments.

Coaxial Triboelectric Nanogenerator and Supercapacitor Fiber-Based Self-Charging Power Fabric.

Although there has been rapid advancement in wearable electronics, challenges still remain in developing wearable and sustainable power sources with simple fabrication and low cost. In this work, we demonstrate a flexible coaxial fiber by fabricating a one-dimensional triboelectric nanogenerator (TENG) outside and a supercapacitor (SC) inside, which can not only harvest mechanical energy but also store energy in the all-in-one fiber. In such a coaxial fiber, carbon fiber bundles are utilized as the electrode material for the TENG as well as the active and electrode material for the SC. Meanwhile, silicone rubber serves as the separator between the SC and TENG, as the triboelectric material for the TENG, and as the encapsulation material for the whole fiber as well. Moreover, both SC and TENG exhibit good performance and stability, which ensures their long-term use in daily life. Because of the flexibility and durability of the carbon fiber and silicone rubber, the proposed coaxial fibers show great flexibility, which could be further knitted as cloth for sustainably powering wearable electronic devices. This work presents a promising platform for wearable electronics as well as smart textiles.

PbS Quantum Dots/2D Nonlayered CdS xSe1- x Nanosheet Hybrid Nanostructure for High-Performance Broadband Photodetectors.

Two-dimensional (2D) nonlayered nanomaterials have attracted extensive attention for electronic and optoelectronic applications recently because of their distinct properties. In this work, we first employed a facile one-step method to synthesize 2D nonlayered cadmium sulfide selenide (CdS xSe1- x, x = 0.33) nanosheets with a highly crystalline structure and then we introduced a generic spin-coating approach to fabricate hybrid nanomaterials composed of PbS quantum dots (QDs) and 2D CdS xSe1- x nanosheets and demonstrated their potential for high-performance broadband photodetectors. Compared with pure 2D CdS xSe1- x nanosheet photodetectors, the photoelectric performance of the PbS/CdS xSe1- x hybrid nanostructure is enhanced by 3 orders of magnitude under near-infrared (NIR) light illumination and maintains its performance in the visible (Vis) range. The photodetector exhibits a broadband response range from Vis to NIR with an ultrahigh light-to-dark current ratio (3.45 × 106), a high spectral responsivity (1.45 × 103 A/W), and high detectivity (1.05 × 1015 Jones). The proposed QDs/2D nonlayered hybrid nanostructure-based photodetector paves a promising way for next-generation high-performance broadband optoelectronic devices.

Loading across the Periodic Table: Introducing 14 Different Metal Ions To Enhance Metal-Organic Framework Performance.

Loading metal guests within metal-organic frameworks (MOFs) via secondary functional groups is a promising route for introducing or enhancing MOF performance in various applications. In this work, 14 metal ions (Li+, Na+, K+, Mg2+, Ca2+, Ba2+, Zn2+, Co2+, Mn2+, Ag+, Cd2+, La3+, In3+, and Pb2+) have been successfully introduced within the MIL-121 MOF using a cost-efficient route involving free carboxylic groups on the linker. The local and long-range structure of the metal-loaded MOFs is characterized using multinuclear solid-state NMR and X-ray diffraction methods. Li/Mg/Ca-loaded MIL-121 and Ag nanoparticle-loaded MIL-121 exhibit enhanced H2 and CO2 adsorption; Ag nanoparticle-loaded MIL-121 also demonstrates remarkable catalytic activity in the reduction of 4-nitrophenol.

Triboelectric Nanogenerator Driven Self-Powered Photoelectrochemical Water Splitting Based on Hematite Photoanodes.

Hematite is one of the most promising photoanodes for photoelectrochemical (PEC) solar water splitting. However, due to the low conduction band position for water reduction, an external bias is necessarily required with the consumption of extra power. In this work, a titanium modified hematite (Ti-Fe2O3) photoanode-based self-powered PEC water splitting system in tandem with a rotatory disc-shaped triboelectric nanogenerator (RD-TENG) has been developed. It is a fantastic strategy to effectively drive the hematite-based PEC water splitting by using the environmental mechanical energy through a TENG. When the rotation speed is 65 rpm (water flowing rate ∼0.61 m/s), the peak current reaches to 0.12 mA under illumination contrast to that in the dark with almost zero. As for 80 rpm, the peak currents are 0.17 and 0.33 mA in the dark or under illumination, respectively, indicating the simultaneous occurrence of electrolysis and PEC water splitting. When higher than 120 rpm, the peak current in the dark is nearly equal to that under illumination, which can be attributed to the high enough peak voltage for direct electrolysis of water. Such a self-powered PEC water splitting system provides an alternative strategy that enables to convert both solar and mechanical energies into chemical energies.

Liquid-Metal-Based Super-Stretchable and Structure-Designable Triboelectric Nanogenerator for Wearable Electronics.

The rapid advancement of intelligent wearable electronics imposes the emergent requirement for power sources that are deformable, compliant, and stretchable. Power sources with these characteristics are difficult and challenging to achieve. The use of liquid metals as electrodes may provide a viable strategy to produce such power sources. In this work, we propose a liquid-metal-based triboelectric nanogenerator (LM-TENG) by employing Galinstan as the electrode and silicone rubber as the triboelectric and encapsulation layer. The small Young's modulus of the liquid metal ensures the electrode remains continuously conductive under deformations, stretching to a strain as large as ∼300%. The surface oxide layer of Galinstan effectively prevents the liquid Galinstan electrode from further oxidization and permeation into silicone rubber, yielding outstanding device stability. Operating in the single-electrode mode at 3 Hz, the LM-TENG with an area of 6 × 3 cm2 produces an open-circuit voltage of 354.5 V, transferred short-circuit charge of 123.2 nC, short-circuit current of 15.6 µA, and average power density of 8.43 mW/m2, which represent outstanding performance values for TENGs. Further, the LM-TENG maintains stable performance under various deformations, such as stretching, folding, and twisting. LM-TENGs in different forms, such as bulk-shaped, bracelet-like, and textile-like, are all able to harvest mechanical energy from human walking, arm shaking, or hand patting to sustainably drive wearable electronic devices.

Constructing tailored isoprenoid products by structure-guided modification of geranylgeranyl reductase.

The archaeal enzyme geranylgeranyl reductase (GGR) catalyzes hydrogenation of carbon-carbon double bonds to produce the saturated alkyl chains of the organism's unusual isoprenoid-derived cell membrane. Enzymatic reduction of isoprenoid double bonds is of considerable interest both to natural products researchers and to synthetic biologists interested in the microbial production of isoprenoid drug or biofuel molecules. Here we present crystal structures of GGR from Sulfolobus acidocaldarius, including the structure of GGR bound to geranylgeranyl pyrophosphate (GGPP). The structures are presented alongside activity data that depict the sequential reduction of GGPP to H6GGPP via the intermediates H2GGPP and H4GGPP. We then modified the enzyme to generate sequence variants that display increased rates of H6GGPP production or are able to halt the extent of reduction at H2GGPP and H4GGPP. Crystal structures of these variants not only reveal the structural bases for their altered activities; they also shed light onto the catalytic mechanism employed.

Crystal structure and biochemical studies of the trans-acting polyketide enoyl reductase LovC from lovastatin biosynthesis.

Lovastatin is an important statin prescribed for the treatment and prevention of cardiovascular diseases. Biosynthesis of lovastatin uses an iterative type I polyketide synthase (PKS). LovC is a trans-acting enoyl reductase (ER) that specifically reduces three out of eight possible polyketide intermediates during lovastatin biosynthesis. Such trans-acting ERs have been reported across a variety of other fungal PKS enzymes as a strategy in nature to diversify polyketides. How LovC achieves such specificity is unknown. The 1.9-Å structure of LovC reveals that LovC possesses a medium-chain dehydrogenase/reductase (MDR) fold with a unique monomeric assembly. Two LovC cocrystal structures and enzymological studies help elucidate the molecular basis of LovC specificity, define stereochemistry, and identify active-site residues. Sequence alignment indicates a general applicability to trans-acting ERs of fungal PKSs, as well as their potential application to directing biosynthesis.

Functional characterization of four sesquiterpene synthases from Ricinus communis (castor bean).

Genome sequence analysis of Ricinus communis has indicated the presence of at least 22 putative terpene synthase (TPS) genes, 13 of which appear to encode sesquiterpene synthases (SeTPSs); however, no SeTPS genes have been isolated from this plant to date. cDNAs were recovered for six SeTPS candidates, and these were subjected to characterization in vivo and in vitro. The RcSeTPS candidates were expressed in either Escherichia coli or Saccharomyces cerevisiae strains with engineered sesquiterpene biosynthetic pathways, but only two (RcSeTPS1 and RcSeTPS7) produced detectable levels of product. In order to check whether the engineered microbial hosts were adequately engineered for sesquiterpene production, a selection of SeTPS genes was chosen from other plant species and demonstrated consistently high sesquiterpene titers. Activity could be demonstrated in vitro for two of the RcSeTPS candidates (RcSeTPS5 and RcSeTPS10) that were not observed to be functional in our microbial hosts. RcSeTPS1 produced two products, (-)-α-copaene and (+)-δ-cadinene, while RcSeTPS7 produced a single product, (E, E)-α-farnesene. Both RcSeTPS5 and RcSeTPS10 produced multiple sesquiterpenes.

FT-ICR-MS characterization of intermediates in the biosynthesis of the α-methylbutyrate side chain of lovastatin by the 277 kDa polyketide synthase LovF.

There are very few fungal polyketide synthases that have been characterized by mass spectrometry. In this paper we describe the in vitro reconstitution and FT-ICR-MS verification of the full activity of an intact 277 kDa fungal polyketide synthase LovF of the lovastatin biosynthetic pathway. We report here both the verification of the reconstitution of fully functional holo-LovF by using (13)C-labeled malonyl-CoA to form α-methylbutyrate functionality and also detection of five predicted intermediates covalently bound to the 4'-phosphopantetheine at the acyl carrier protein (ACP) active site utilizing the phosphopantetheine ejection assay and high-resolution mass spectrometry. Under in vitro conditions, the diketide acetoacetyl intermediate did not accumulate on the ACP active site of holo-LovF following incubation with malonyl-CoA substrate. We found that incubation of holo-LovF with acetoacetyl-CoA served as an effective means of loading the diketide intermediate onto the ACP active site of LovF. Our results demonstrate that subsequent α-methylation of the acetoacetyl intermediate stabilizes the intermediate onto the ACP active site and facilitates the formation and mass spectrometric detection of additional intermediates en route to the formation of α-methylbutyrate.

Complete reconstitution of a highly reducing iterative polyketide synthase.

Highly reducing iterative polyketide synthases are large, multifunctional enzymes that make important metabolites in fungi, such as lovastatin, a cholesterol-lowering drug from Aspergillus terreus. We report efficient expression of the lovastatin nonaketide synthase (LovB) from an engineered strain of Saccharomyces cerevisiae, as well as complete reconstitution of its catalytic function in the presence and absence of cofactors (the reduced form of nicotinamide adenine dinucleotide phosphate and S-adenosylmethionine) and its partner enzyme, the enoyl reductase LovC. Our results demonstrate that LovB retains correct intermediates until completion of synthesis of dihydromonacolin L, but off-loads incorrectly processed compounds as pyrones or hydrolytic products. Experiments replacing LovC with analogous MlcG from compactin biosynthesis demonstrate a gate-keeping function for this partner enzyme. This study represents a key step in the understanding of the functions and structures of this family of enzymes.

Directed evolution and structural characterization of a simvastatin synthase.

Enzymes from natural product biosynthetic pathways are attractive candidates for creating tailored biocatalysts to produce semisynthetic pharmaceutical compounds. LovD is an acyltransferase that converts the inactive monacolin J acid (MJA) into the cholesterol-lowering lovastatin. LovD can also synthesize the blockbuster drug simvastatin using MJA and a synthetic alpha-dimethylbutyryl thioester, albeit with suboptimal properties as a biocatalyst. Here we used directed evolution to improve the properties of LovD toward semisynthesis of simvastatin. Mutants with improved catalytic efficiency, solubility, and thermal stability were obtained, with the best mutant displaying an approximately 11-fold increase in an Escherichia coli-based biocatalytic platform. To understand the structural basis of LovD enzymology, seven X-ray crystal structures were determined, including the parent LovD, an improved mutant G5, and G5 cocrystallized with ligands. Comparisons between the structures reveal that beneficial mutations stabilize the structure of G5 in a more compact conformation that is favorable for catalysis.

Acyltransferase mediated polyketide release from a fungal megasynthase.

LovF is a highly reducing polyketide synthase (HR-PKS) from the filamentous fungus Aspergillus terreus. LovF synthesizes the alpha-S-methylbutyrate side chain that is subsequently transferred to monacolin J to yield the cholesterol-lowering natural product lovastatin. In the report, we expressed the full length LovF and reconstituted the megasynthase activities in vitro. We confirmed the diketide product of LovF is offloaded from the LovF ACP domain by the dissociated acyltransferase LovD. This represents the first example of acyltransferase-mediated release of polyketide products from fungal PKSs. We determined LovD primarily interacts with the ACP domain of LovF and the protein-protein interactions lead to highly efficient transfer of the diketide product. The catalytic efficiency is enhanced nearly 1 x 10(6)-fold when LovF was used as the acyl carrier instead of N-acetylcysteamine. Reconstitution and characterization of the LovF offloading mechanism provide new insights into the functions of fungal HR-PKS.

Rational improvement of simvastatin synthase solubility in Escherichia coli leads to higher whole-cell biocatalytic activity.

Simvastatin is the active pharmaceutical ingredient of the blockbuster cholesterol lowering drug Zocor. We have previously developed an Escherichia coli based whole-cell biocatalytic platform towards the synthesis of simvastatin sodium salt (SS) starting from the precursor monacolin J sodium salt (MJSS). The centerpiece of the biocatalytic approach is the simvastatin synthase LovD, which is highly prone to misfolding and aggregation when overexpressed from E. coli. Increasing the solubility of LovD without decreasing its catalytic activity can therefore elevate the performance of the whole-cell biocatalyst. Using a combination of homology structural prediction and site-directed mutagenesis, we identified two cysteine residues in LovD that are responsible for nonspecific intermolecular crosslinking, which leads to oligomer formation and protein aggregation. Replacement of Cys40 and Cys60 with alanine residues resulted in marked gain in both protein solubility and whole-cell biocatalytic activities. Further mutagenesis experiments converting these two residues to small or polar natural amino acids showed that C40A and C60N are the most beneficial, affording 27% and 26% increase in whole cell activities, respectively. The double mutant C40A/C60N combines the individual improvements and displayed approximately 50% increase in protein solubility and whole-cell activity. Optimized fed-batch high-cell-density fermentation of the double mutant in an E. coli strain engineered for simvastatin production quantitatively (>99%) converted 45 mM MJSS to SS within 18 h, which represents a significant improvement over the performance of wild-type LovD under identical conditions. The high efficiency of the improved whole-cell platform renders the biocatalytic synthesis of SS an attractive substitute over the existing semisynthetic routes.

Engineering natural products using combinatorial biosynthesis and biocatalysis.

Many biologically active natural products are produced by the host organisms using dedicated biosynthetic pathways. The programming rules of these pathways may be rationally manipulated through combinatorial biosynthesis to produce natural products that contain structural variations or enhanced pharmacological properties. Furthermore, these pathways contain enzymes that can be harvested as powerful biocatalysts for the synthesis of both new drugs and existing blockbuster therapeutics. This review will highlight recent advances in exploring natural product biosynthetic pathways for new compounds, novel enzymes and useful biocatalysts.

A polyketide macrolactone synthase from the filamentous fungus Gibberella zeae.

Resorcylic acid lactones represent a unique class of fungal polyketides and display a wide range of biological activities, such as nanomolar inhibitors of Hsp90 and MAP kinase. The biosynthesis of these compounds is proposed to involve two fungal polyketide synthases (PKS) that function collaboratively to yield a 14-membered macrolactone with a resorcylate core. We report here the reconstitution of Gibberella zeae PKS13, which is the nonreducing PKS associated with zearalenone biosynthesis. Using a small molecule mimic of the natural hexaketide starter unit, we reconstituted the entire repertoire of PKS13 activities in vitro, including starter-unit selection, iterative condensation, regioselective C2-C7 cyclization, and macrolactone formation. PKS13 synthesized both natural 14-membered and previously uncharacterized 16-membered resorcylic acid lactones, indicating relaxed control in both iterative elongation and macrocyclization. PKS13 exhibited broad starter-unit specificities toward fatty acyl-CoAs ranging in sizes between C6 and C16 and displayed the highest activity toward decanoyl-CoA. PKS13 was shown to be active in Escherichia coli and synthesized numerous alkyl pyrones and alkyl resorcylic esters without exogenously supplied precursors. We demonstrated that PKS13 can interact with E. coli fatty acid biosynthetic machinery and can be primed with fatty-acyl ACPp at low-micromolar concentrations. PKS13 synthesized new polyketides in E. coli when the culture was supplemented with synthetic precursors, showcasing its utility in precursor-directed biosynthesis. PKS13 is therefore a highly versatile polyketide macrolactone synthase that is useful in the engineered biosynthesis of polyketides, including resorcylic acid lactones that are not found in nature.

Redirecting the cyclization steps of fungal polyketide synthase.

Regiospecific cyclizations of the nascent poly-beta-ketone backbones dictate the structures of polyketide natural products. The fungal iterative megasynthases use terminal thioesterase/claisen cyclase (TE/CLC) domains to direct the fate of the polyketide chains. In this work, we present two strategies toward redirecting the cyclization steps of fungal PKSs using the Gibberella fujikuroi PKS4. First, inactivation or removal of the TE/CLC domain resulted in the synthesis of the new polyketide SMA93 2. Complementation of the mutant PKS4 with a standalone TE/CLC domain restored the regioselective cyclization steps of PKS4 and led to the synthesis of SMA76 1, demonstrating that cyclization enzymes can interact with the megasynthase in trans. This led to the second approach in which various dissociated, bacterial tailoring enzymes were added to the megasynthase in trans. Addition of the act KR led to the synthesis of mutactin 3, while the addition of first ring and second ring cyclases yielded anthraquinone compounds DMAC 5 and SEK26 6. The cooperative activities of fungal and bacterial PKS components are especially important and enable synthesis of polyketides utilizing enzymes from two distinct families of PKSs.

Improving simvastatin bioconversion in Escherichia coli by deletion of bioH.

Simvastatin is an important cholesterol lowering compound and is currently synthesized from the natural product lovastatin via multistep chemical synthesis. We have previously reported the use of an Escherichia coli strain BL21(DE3)/pAW31 as the host for whole-cell biocatalytic conversion of monacolin J acid to simvastatin acid. During fermentation and bioconversion, unknown E. coli enzyme(s) hydrolyzed the membrane permeable thioester substrate dimethylbutyryl-S-methyl mercaptopropionate (DMB-S-MMP) to the free acid, significantly decreased the efficiencies of the whole-cell bioconversion and the downstream purification steps. Using the Keio K-12 Singe-Gene Knockout collection, we identified BioH as the sole enzyme responsible for the observed substrate hydrolysis. Purification and reconstitution of E. coli BioH activity in vitro confirmed its function. BioH catalyzed the rapid hydrolysis of DMB-S-MMP with kcat and Km values of 260+/-45 s(-1) and 229+/-26 microM, respectively. This is in agreement with previous reports that BioH can function as a carboxylesterase towards fatty acid esters. YT2, which is a delta bioH mutant of BL21(DE3), did not hydrolyze DMB-S-MMP during prolonged fermentation and was used as an alternative host for whole-cell biocatalysis. The rate of simvastatin acid synthesis in YT2 was significantly faster than in BL21(DE3) and 99% conversion of 15 mM simvastatin acid in less than 12 h was achieved. Furthermore, the engineered host required significantly less DMB-S-MMP to be added to accomplish complete conversion. Finally, simvastatin acid synthesized using YT2 can be readily purified from fermentation broth and no additional steps to remove the hydrolyzed dimethylbutyryl-S-mercaptopropionic acid is required. Together, the proteomic and metabolic engineering approaches render the whole-cell biocatalytic process more robust and economically attractive.