Two-step Growth Research Articles

Combining culture optimization and synthetic biology to improve production and detection of secondary metabolites in Myxococcus xanthus: application to myxoprincomide.

Microbial secondary metabolites play crucial ecological roles in governing species interactions and contributing to their defense strategies. Their unique structures and potent bioactivities have been key in discovering antibiotics and anticancer drugs. Genome sequencing has undoubtedly revealed that myxobacteria constitute a huge reservoir of secondary metabolites as the well-known producers, actinomycetes. However, because most secondary metabolites are not produced in the laboratory context, the natural products from myxobacteria characterized to date represent only the tip of the iceberg. By combining the engineering of a dedicated Myxococcus xanthus DZ2 chassis strain with a two-step growth medium protocol, we provide a new approach called two-step Protocol for Resource Integration and Maximization-Biomolecules Overproduction and Optimal Screening Therapeutics (2PRIM-BOOST) for the production of non-ribosomal peptides synthetases (NRPS)/polyketides synthases (PKS) secondary metabolites from myxobacteria. We further show that the 2PRIM-BOOST strategy will facilitate the screening of secondary metabolites for biological activities of medical interest. As proof of concept, using a constitutive strong promoter, the myxoprincomide from M. xanthus DZ2 has been efficiently produced and its biosynthesis has been enhanced using the 2PRIM-BOOST approach, allowing the identification of new features of myxoprincomide. This strategy should allow the chances to produce and discover new NRPS, PKS, and mixed NRPS/PKS hybrid natural metabolites that are currently considered as cryptic and are the most represented in myxobacteria.IMPORTANCEMicrobial secondary metabolites are important in species interactions and are also a prolific source of drugs. Myxobacteria are ubiquitous soil-dwelling bacteria constituting a huge reservoir of secondary metabolites. However, because most of these molecules are not produced in the laboratory context, one can estimate that only one-tenth have been characterized to date. Here, we developed a new strategy called two-step Protocol for Resource Integration and Maximization-Biomolecules Overproduction and Optimal Screening Therapeutics (2PRIM-BOOST) that combines the engineering of a dedicated Myxococcus xanthus chassis strain together with growth medium optimization. By combining these strategies with the insertion of a constitutive promoter upstream the biosynthetic gene cluster (BGC), the production of myxoprincomide, a characterized low-produced secondary metabolite, was successfully and significantly increased. The 2PRIM-BOOST enriches the toolbox used to produce previously cryptic metabolites, unveil their ecological role, and provide new molecules of medical interest.

Two-step nucleation and growth of crystals in a metastable solution with mass exchange with the environment

Two-step nucleation and growth of crystals in a metastable solution with mass exchange with the environment

Sulfur fixation performance of calcium-bearing carbonates in iron ore roasting process: Thermodynamics, phase transition, microstructure evolution, and kinetics

Roasting technology is increasingly used in the beneficiation of refractory iron ores. During the roasting process, the presence of pyrite in some iron ores leads to the formation of SO2, requiring effective sulfur fixation techniques. This study proposed an innovative in-situ sulfur fixation method by pre-oxidation roasting, in which sulfur was fixed during the oxidation roasting process before reduction roasting. The effects of the main iron mineral (hematite), primary gangue mineral (quartz), and common calcium-containing carbonate minerals (calcite and dolomite) on sulfur migration during pyrite roasting were investigated, with particular emphasis on the sulfur fixation capabilities of calcite and dolomite. The study included thermodynamic equilibrium analysis, phase transitions and microstructural evolution in different reaction systems. The results showed that calcite and dolomite effectively reduced SO2 emissions by forming CaSO4 and MgSO4, with calcite exhibiting a stronger sulfur fixation capacity for the same mass. Kinetic analysis indicated that pyrite pyrolysis followed a two-step random nucleation and growth model. Furthermore, the addition of calcite increased the apparent activation energy of SO2 formation and altered the reaction pathway, providing insight into the sulfur fixation mechanism of calcite from a kinetic perspective.

Characterisation of growth parameters for the extremely acidophilic archaeon Ferroplasma acidiphilum DSM 12658 using a two-step turbidimetric growth measurement method

The acidophilic archaeon Ferroplasma acidiphilum exhibits remarkable adaptations to life in highly acidic environments. Nevertheless, the investigation of its molecular biology is challenging because of the slow growth of the organism, low biomass yield, and limitations of standard growth measurement techniques caused by iron oxidation.•A novel two-step turbidimetric growth measurement (2TGM) method was developed to address the shortcomings associated with iron precipitation in culture media. The method comprises two distinct preparation steps.•The initial 2TGM step involves centrifuging culture samples at high speed to remove F. acidiphilum cells and iron precipitates, thus preparing a blank.•The second 2TGM step involves centrifuging the culture at a lower speed with the objective of selectively removing iron precipitates, thus allowing F. acidiphilum cells to remain suspended in the supernatant. This preparation enables subsequent measurement of cell density.A study on the growth of F. acidiphilum showed a standard logistic pattern with a 35-h lag phase, approximately 9-h doubling time, and 0.042 OD carrying capacity. A new two-step turbidimetric growth measurement (2TGM) method overcomes limitations of existing approaches, enabling further investigation of F. acidiphilum.

Development of GeSn epitaxial films with strong direct bandgap luminescence in the mid-wave infrared region using a commercial chemical vapor deposition reactor

Germanium tin (GeSn) is a material of interest for electronic and photonic device applications, but its development and commercialization have been limited by material quality issues and lack of availability from epitaxy suppliers. In this paper, we report initial studies in optimizing GeSn films deposited on a Ge buffer layer grown on 200-mm diameter silicon (Si) substrates with an ASM Epsilon 2000 chemical vapor deposition reactor designed for commercial production. Using a single-step growth process, a Sn content up to 22% near the surface of a GeSn film was achieved due to the increase in Sn incorporation via strain relaxation. A two-step growth process resulted in a bilayer structure with a nearly 100% relaxation on the first layer, followed by a higher quality GeSn layer with 18% Sn as evident by a high photoluminescence intensity emitting in the mid-wave infrared region at 3.2 μm at 20 K.

Two-step crystallization in a supersaturated solution with application to protein crystal growth: Theory and analytical solutions

A theory of two-step nucleation and evolution of crystals with allowance for their diffusion in the space of particle radii is developed. The theory is based on a two-step growth law of crystals appearing in dense liquid precursors, initially evolving within them with a diffusion-limited growth law, and then leaving them and evolving with another growth law. This two-step crystal growth law influences the integrodifferential system of kinetic and balance equations for the crystal-size distribution function and liquid supersaturation. The system of integrodifferential equations for the evolution of polydisperse ensemble of crystals is solved using the integral Laplace transform and saddle-point methods for the Weber–Volmer–Frenkel–Zel’dovich and Meirs kinetic mechanisms. A complete analytical solution to this non-linear system has been constructed in a parametric form. Namely, the particle-size distribution function, liquid supersaturation and time have been found as the functions of decision variable — the maximum size of crystals. The theory is in agreement with the experimental data on the growth of beta-lactoglobulin and insulin crystals.

Seed-assisted two-step ZIF-67 growth on CS/PVA nanofibers for high-efficiency cadmium and tetracycline adsorption

Herein, novel ZIF-67/CS/PVA (CNF-2) nanofiber composites using a two-step seed-assisted in situ electrospinning method, with cobalt-based zeolitic imidazolate framework (ZIF-67) as the precursor was prepared. SEM, EDX, AFM, FTIR, and XRD analyses revealed the optimal formation of ZIF-67 nanoparticles both inside and on the surface of CS/PVA nanofibers without compromising the structure of these electrospun nanofibers. The unique porous structure and heterointerfaces of the ZIF-67/CS/PVA (CNF-2) nanofiber composites demonstrated remarkable adsorption capacities for cadmium (Cd) and tetracycline hydrochloride (TCH), achieving 1137.94 and 1029.5 mg/g, respectively. The adsorption effectiveness for Cd and TCH was 94.83 % and 85.79 %, respectively, under optimal adsorption conditions: pH 8, adsorbent dosage of 0.01 g, initial Cd and TCH concentration of 80 mg/L, and a contact time of 120 min. The study of coexisting Cd and TCH adsorption on CNF-2 under varying pH conditions (4–8) reveals a significant enhancement in Cd adsorption efficiency, increasing from 41.91 % to 97.49 %. Conversely, TCH exhibits a more modest improvement, with its adsorption efficiency rising from 72.53 % to 85.96 %. Kinetic and equilibrium studies revealed that the adsorption followed pseudo-second-order kinetics and the Langmuir isotherm. The prepared nanofiber adsorbed 60 % of Malachite green dye. The engineered adsorbent demonstrated impressive regeneration potential, maintaining its efficiency over three successive adsorption–desorption cycles.

Geometrically Controlled Microscale Patterning and Epitaxial Lateral Overgrowth of Nitrogen-Polar GaN.

In this Letter, we report on a novel two-step epitaxial growth technique that enables a significant improvement of the crystal quality of nitrogen-polar GaN. The starting material is grown on 4° vicinal sapphire substrates by metal-organic vapor-phase epitaxy, with an initial high-temperature sapphire nitridation to control polarity. The material is then converted to a regular array of hexagonal pyramids by wet-etching in a KOH solution and subsequently regrown to coalesce the pyramids back into a smooth layer of improved crystal quality. The key points that enable this technique are the control of the array geometry, obtained by exploiting the anisotropic behavior of the wet-etch step, and the use of regrowth conditions that preserve the orientation of the pyramids' sidewalls. In contrast, growth conditions that cause an excessive expansion of the residual (0001̅) facets on the pyramids' tops cause the onset of a very rough surface morphology upon full coalescence. An X-ray diffraction study confirms the reduction of the threading dislocation density as the regrowth step develops. The analysis of the relative position of the 0002̅ GaN peak, with respect to the 0006 sapphire peak, reveals a macroscopic tilt of the pyramids, probably induced by the large off-axis substrate orientation. This tilt correlates very well with an anomalous broadening of the 0002̅ diffraction peaks at the beginning of the regrowth step.

Enhanced optical properties of MoSe2 grown by molecular beam epitaxy on hexagonal boron nitride

Transition metal dichalcogenides (TMDs) like MoSe2 exhibit remarkable optical properties such as intense photoluminescence (PL) in the monolayer form. To date, narrow-linewidth PL is only achieved in micrometer-sized exfoliated TMD flakes encapsulated in hexagonal boron nitride (hBN). In this work, we develop a growth strategy to prepare monolayer MoSe2 on hBN flakes by molecular beam epitaxy in the van der Waals regime. It constitutes the first step toward the development of large area single crystalline TMDs encapsulated in hBN for potential integration in electronic or optoelectronic devices. For this purpose, we define a two-step growth strategy to achieve monolayer-thick MoSe2 grains on hBN flakes. The high quality of MoSe2 allows us to detect very narrow PL linewidth down to 5.5 meV at 13 K, comparable to the one of encapsulated exfoliated MoSe2 flakes. Moreover, sizeable PL can be detected at room temperature as well as clear reflectivity signatures of A, B, and charged excitons.

Novel approach to produce 3D boron-doped diamond for pollutant removal from water

Diamond growth from Chemical Vapor Deposition (CVD) on foreign substrates can require different pretreatment not only to improve the film nucleation but also to assure its adhesion by decreasing the expected film/substrate interface stress. To improve boron-doped film nucleation, growth, and adherence, different substrate pretreatments have been used mainly from the seeding process with diamond powder at various particle sizes. Despite this, the development of diamond growth on a Ti mesh remains difficult because of the requirement of a cohesive film to cover a 3D macroporous sample with varying growth rates based on its distinct network geometry. Then, this work describes a novel approach to growing boron-doped diamond (BDD) and boron-doped ultrananocrystalline diamond (B-UNCD) on titanium dioxide nanotubes (TDNT) produced simultaneously on both sides of Ti mesh by an anodization process. The films were obtained from two-step growth processes by assuring the entire diamond overlay on both TDNT/Ti mesh sides, including their outer/inner surfaces, as a 3D sample. TiO2 - TiC conversion has dominated the renucleation process, facilitating the nanometric scale control. The film morphologies were systematically analyzed by FEG-SEM images at different sample planes and depths for both sample sides at different stages of film growth. The unique morphology of titania nanotubes associated with columnar and/or renucleation development of BDD, considering the film defects and valley, can systematically increase the electrode specific area. Raman spectra showed the film quality and its micro and/or ultrananodiamond structure and the boron doping features. Also, this growth process allowed a dopant-controlled adjustable conductivity. Then, the boron doping levels for both films were evaluated from Mott-Schottky plots at around 1019 Bcm−3, characterizing them with good conductivity. In addition, electrochemical measurements from Cyclic Voltammetry (CV) confirmed the expected diamond response on redox pair following the quasi-reversible criteria as high-performance diamond electrodes and in situ Raman spectroelectrochemical measurements assessed the stability of samples during electrochemical measurements, ensuring structural integrity. Finally, the samples were applied to the degradation of methylene blue, proving to be superior materials for electrochemical applications due to their advantages compared to those of similar 2D electrodes.

Large-area and few-layered 1T′-MoTe2 thin films grown by cold-wall chemical vapor deposition

This study employs cold-wall chemical vapor deposition to achieve the growth of MoTe2 thin films on 4-inch sapphire substrates. A two-step growth process is utilized, incorporating MoO3 and Te powder sources under low-pressure conditions to synthesize MoTe2. The resultant MoTe2 thin films exhibit a dominant 1T′ phase, as evidenced by a prominent Raman peak at 161 cm−1. This preferential 1T′ phase formation is attributed to controlled manipulation of the second-step growth temperature, essentially the reaction stage between Te vapor and the pre-deposited MoO x layer. Under these optimized growth conditions, the thickness of the continuous 1T′-MoTe2 films can be precisely tailored within the range of 3.5–5.7 nm (equivalent to 5–8 layers), as determined by atomic force microscopy depth profiling. Hall-effect measurements unveil a typical hole concentration and mobility of 0.2 cm2 Vs−1 and 7.9 × 1021 cm−3, respectively, for the synthesized few-layered 1T′-MoTe2 films. Furthermore, Ti/Al bilayer metal contacts deposited on the few-layered 1T′-MoTe2 films exhibit low specific contact resistances of approximately 1.0 × 10−4 Ω cm2 estimated by the transfer length model. This finding suggests a viable approach for achieving low ohmic contact resistance using the 1T′-MoTe2 intermediate layer between metallic electrodes and two-dimensional semiconductors.

Nucleation of InP on Si under micro-crucibles at ultra-high vacuum using a two-step VLS process

We reported nucleation mechanisms of InP directly on Si (8% lattice mismatch) under confined structures, called micro-crucibles, at ultra-high vacuum (UHV) by chemical beam epitaxy. These micro-crucibles are used to induce lateral growth in the presence of a micro-scale Au catalyst. It is found that at this UHV condition, the kinetics is dictated predominantly by adatom surface diffusion. Using a two-step growth process ((1) In-only exposure, then, (2) simultaneous In and P exposures), InP islands have been successfully nucleated on Si substrates under micro-crucible structures. The nucleation of these InP islands strongly depends on the metal catalyst location relative to the micro-crucible opening with metal catalysts residing closer to the opening having a higher chance to get incorporated with In and P atoms. Importantly, we found that using smaller micro-crucibles with double openings can increase the possibility of having metal catalysts reside near either opening and nucleate InP under micro-crucibles.

Growth of Monolayer MoS2 Flakes via Close Proximity Re-Evaporation.

We report a two-step growth process of MoS2 nanoflakes using a low-pressure chemical vapor deposition technique. In the first step, a MoS2 layer was synthesized on a c-plane sapphire substrate. This layer was subsequently re-evaporated at a higher temperature to form mono- or few-layer MoS2 flakes. As a result, the close proximity re-evaporation enabled the growth of pristine MoS2 nanoflakes. Atomic force microscopy analysis confirmed the synthesis of nanoclusters/nanoflakes with lateral dimensions of over 10 μm and a flake height of approximately 1.3 nm, demonstrating bi-layer MoS2, whereas transmission electron microscopy analysis revealed triangular MoS2 nanoflakes, with a diffraction pattern proving the presence of single crystalline hexagonal MoS2. Raman data revealed the typical modes of high-quality MoS2 nanoflakes. Finally, we presented the photocurrent dependence of a MoS2-based photoresist under illumination with light-emitting diode of 405 nm wavelength. The measured current-voltage dependence across various luminous flux outlined the sensitivity of MoS2 to polarized light and thus opens further opportunities for applications in high-performance photodetectors with polarization sensitivity.

Into the Void: Single Nanopore in Colloidally Synthesized Bi2Te3 Nanoplates with Ultralow Lattice Thermal Conductivity.

Bi2Te3 is a well-known thermoelectric material that was first investigated in the 1960s, optimized over decades, and is now one of the highest performing room-temperature thermoelectric materials to-date. Herein, we report on the colloidal synthesis, growth mechanism, and thermoelectric properties of Bi2Te3 nanoplates with a single nanopore in the center. Analysis of the reaction products during the colloidal synthesis reveals that the reaction progresses via a two-step nucleation and epitaxial growth: first of elemental Te nanorods and then the binary Bi2Te3 nanoplate growth. The rates of epitaxial growth can be controlled during the reaction, thus allowing the formation of a single nanopore in the center of the Bi2Te3 nanoplates. The size of the nanopore can be controlled by changing the pH of the reaction solution, where larger pores with diameter of ∼50 nm are formed at higher pH and smaller pores with diameter of ∼16 nm are formed at lower pH. We propose that the formation of the single nanopore is mediated by the Kirkendall effect and thus the reaction conditions allow for the selective control over pore size. Nanoplates have well-defined hexagonal facets as seen in the scanning and transmission electron microscopy images. The single nanopores have a thin amorphous layer at the edge, revealed by transmission electron microscopy. Thermoelectric properties of the pristine and single-nanopore Bi2Te3 nanoplates were measured in the parallel and perpendicular directions. These properties reveal strong anisotropy with a significant reduction to thermal conductivity and increased electrical resistivity in the perpendicular direction due to the higher number of nanoplate and nanopore interfaces. Furthermore, Bi2Te3 nanoplates with a single nanopore exhibit ultralow lattice thermal conductivity values, reaching ∼0.21 Wm-1K-1 in the perpendicular direction. The lattice thermal conductivity was found to be systematically lowered with pore size, allowing for the realization of a thermoelectric figure of merit, zT of 0.75 at 425 K for the largest pore size.

Amorphous-to-crystalline transition-induced two-step thin film growth of quasi-one-dimensional penta-telluride ZrTe5

Quasi-one-dimensional (quasi-1D) van der Waals (vdWs) materials, such as ZrTe5, exhibit unique electrical properties and quantum phenomena, making them attractive for advanced electronic applications. However, large-scale growth of ZrTe5 thin films presents challenges. We address this by employing sputtering, a common semiconductor industry technique. The as-deposited ZrTe5 film is amorphous, and post-annealing induces a crystallization process akin to transition-metal dichalcogenides. Our study investigates the electrical and optical properties during this amorphous-to-crystalline transition, revealing insights into the underlying mechanism. This work contributes to the fundamental understanding of quasi-1D materials and introduces a scalable fabrication method for ZrTe5 which offers the possibility of fabricating unique future electronic and optical devices.

Unraveling Halogen Role in Two-Step Solution Growth of Organic-Inorganic Hybrid Mixed-Halide Perovskites: Guidelines of Fabricating Single-Phase Perovskites with Predictable Stoichiometry.

The challenge faced in optoelectronic applications of halide perovskites is their degradation. Minimizing material imperfections is critical to averting cascade degradation processes. Identifying causes of such imperfections is, however, hindered by mystified growth processes and is particularly urgent for mixed-halide perovskites because of inhomogeneity in growth and phase segregation under stresses. To unravel two-step solution growth of MAPbBr x I3-x , we monitored the evolution of Br composition and found that the construction of perovskite lattice is contributed by iodine from PbI2 substrate and Br from MABr solution with a 1:1 ratio rather than a 2:1 ratio originally thought. Kinetic analysis based on a derived three-stage model extracted activation energies of perovskite construction and anion exchange. This model is applicable to the growth of PbI2 reacting with a mixed solution of MABr and MAI. Two guidelines of fabricating single-phase MAPbBr x I3-x with predictable stoichiometry thus developed help strategizing protocols to reproducibly fabricate mixed-halide perovskite films tailored to specific optoelectronic applications.

Growth of Quasi-Two-Dimensional CrTe Nanoflakes and CrTe/Transition Metal Dichalcogenide Heterostructures

Two-dimensional (2D) van der Waals layered materials have been explored in depth. They can be vertically stacked into a 2D heterostructure and represent a fundamental way to explore new physical properties and fabricate high-performance nanodevices. However, the controllable and scaled growth of non-layered quasi-2D materials and their heterostructures is still a great challenge. Here, we report a selective two-step growth method for high-quality single crystalline CrTe/WSe2 and CrTe/MoS2 heterostructures by adopting a universal CVD strategy with the assistance of molten salt and mass control. Quasi-2D metallic CrTe was grown on pre-deposited 2D transition metal dichalcogenides (TMDC) under relatively low temperatures. A 2D CrTe/TMDC heterostructure was established to explore the interface’s structure using scanning transmission electron microscopy (STEM), and also demonstrate ferromagnetism in a metal–semiconductor CrTe/TMDC heterostructure.

Investigation of hydrothermally-produced ZnO nanorods and the mechanisms of Li incorporation as a possible dopant

Zinc oxide (ZnO) has emerged as one of the most promising candidates for mass-producing cost-efficient optoelectronic devices. This is primarily because it can be synthesized in high-quality nanostructures on a wide range of substrates through relatively simple chemical methods. However, producing p-type ZnO, regardless of the chosen method, remains an open and controversial issue. In this work, Li-doped ZnO nanostructures of varying Li-cocnentration were produced via a two-step hydrothermal growth synthesis and an in-depth analysis based on with Field Emission Scanning Electron Microscopy (FE-SEM), X-ray diffraction (XRD), Raman Spectroscopy, Extended X-Ray Absorption Fine Structure (EXAFS) Spectroscopy, and temperature-dependent Photoluminescence (PL) was carried out in an effort to gain insights into the Li-incorporation mechanisms. The findings indicated a strong interplay between the native defects responsible for the inherent n-type character of the material and Li incorporation. It is suggested that this interplay hinders the successful conversion of the Li-doped nanorods into p-type nanostructures and that when employing the hydrothermal approach it is essential to identify the precise conditions necessary for genuine Li incorporation as a Zn substitutional.

Two-Step Growth Mechanism of the Solid Electrolyte Interphase in Argyrodyte/Li-Metal Contacts.

The structure and growth of the solid electrolyte interphase (SEI) region between an electrolyte and an electrode is one of the most fundamental yet less well-understood phenomena in solid-state batteries. We present an atomistic simulation of the SEI growth for one of the currently promising solid electrolytes (Li6PS5Cl), based on ab initio-trained machine learning interatomic potentials, for over 30,000 atoms during 10 ns, well beyond the capabilities of conventional molecular dynamics. This unveils a two-step growth mechanism: a Li-argyrodite chemical reaction leading to the formation of an amorphous phase, followed by a kinetically slower crystallization of the reaction products into a 5Li2S·Li3P·LiCl solid solution. The simulation results support the recent, experimentally founded hypothesis of an indirect pathway of electrolyte reduction. These findings shed light on the intricate processes governing SEI evolution, providing a valuable foundation for the design and optimization of next-generation solid-state batteries.

High-quality germanium growth on (111)-faceted V-groove silicon by molecular beam epitaxy

High-quality and low-defect-density germanium (Ge) buffer layers on silicon (Si) substrates have long been developed for group IV and III–V devices by suppressing defect propagation during epitaxial growth. This is a crucial step for the development of highly efficient photonic devices on Si substrates. Patterned silicon substrates have increasingly been employed for their ability to restrict and hinder the motion of defects. In this work, we demonstrate the effectiveness of an optimised two-step growth recipe structure on a (111)-faceted V-groove silicon substrate with a 350 nm flat ridge. This strategy successfully reduces the threading dislocation (TD) density while growing a 1 μm Ge buffer layer via molecular beam epitaxy. As a result, a high-quality buffer is produced with a low TD density on the order of 107 cm−2 and a surface roughness below 1 nm.