Substrate Engineering Research Articles

Breathable and Stretchable Epidermal Electronics for Health Management: Recent Advances and Challenges.

Advanced epidermal electronic devices, capable of real-time monitoring of physical, physiological, and biochemical signals and administering appropriate therapeutics, are revolutionizing personalized healthcare technology. However, conventional portable electronic devices are predominantly constructed from impermeable and rigid materials, which thus leads to the mechanical and biochemical disparities between the devices and human tissues, resulting in skin irritation, tissue damage, compromised signal-to-noise ratio (SNR), and limited operational lifespans. To address these limitations, a new generation of wearable on-skin electronics built on stretchable and porous substrates has emerged. These substrates offer significant advantages including breathability, conformability, biocompatibility, and mechanical robustness, thus providing solutions for the aforementioned challenges. However, given their diverse nature and varying application scenarios, the careful selection and engineering of suitable substrates is paramount when developing high-performance on-skin electronics tailored to specific applications. This comprehensive review begins with an overview of various stretchable porous substrates, specifically focusing on their fundamental design principles, fabrication processes, and practical applications. Subsequently, a concise comparison of various methods is offered to fabricate epidermal electronics by applying these porous substrates. Following these, the latest advancements and applications of these electronics are highlighted. Finally, the current challenges are summarized and potential future directions in this dynamic field are explored.

Extremely Low Thermal Resistance of β-Ga2O3 MOSFETs by Co-integrated Design of Substrate Engineering and Device Packaging.

Gallium oxide (Ga2O3) emerges as a promising ultrawide bandgap semiconductor, which is expected to surpass the performance of current wide bandgap materials, like GaN and SiC, in electronic devices. However, widespread application of Ga2O3 is hindered by its extremely low thermal conductivity and lack of effective device-level thermal management strategies. In this work, Ga2O3 metal-oxide-semiconductor field-effect transistors (MOSFETs) are fabricated by conducting co-integrated design of substrate engineering with layer transferring and device packaging. 3D Raman thermography is introduced as a novel method to analyze the temperature distribution within the device, which provides valuable insights into their thermal performances. A high-quality Ga2O3-SiC heterogeneous integrated material is successfully fabricated with an extremely low interface thermal resistance of 6.67 ± 2 m2·K/GW. Compared to the homoepitaxial Ga2O3 MOSFETs, the degradation of Ion/Ioff in Ga2O3-SiC MOSFETs is decreased by 1.5 orders of magnitude, and that of Ron is decreased by 31%, showing the great thermal stability of Ga2O3-SiC MOSFETs. With the additional device packaging, a significant one order-of-magnitude reduction in the thermal resistance of the Ga2O3-SiC MOSFET is achieved, reaching a record-low value of 4.45 K·mm/W in the reported Ga2O3 MOSFETs. This work demonstrates an efficient strategy for device-level thermal management in next-generation Ga2O3 power and RF applications.

High-Performance perovskite quantum dots light-emitting diodes with hole transport layer engineering and synergetic outcoupling enhancement

Perovskite quantum dot light-emitting diodes (PeQLEDs) are frequently considered as the most promising alternatives to organic light-emitting diodes (OLEDs). However, the efficiency of PeQLEDs remains inferior to OLEDs due to suboptimal charge carrier transport and intrinsic light outcoupling efficiency (ηout). Herein, a combination of hole transport layer (HTL) engineering and substrate engineering is demonstrated to improve the charge injection and ηout of PeQLEDs. To replace the conventional PEDOT:PSS, a novel HTL bilayer based on modified PEDOT:PSS and PVK with a lower refractive index and fewer trap densities at the HTL-QDs interface is developed. Additionally, this study identifies the ideal thickness of ITO for achieving optimal ηout of PeQLEDs through optical simulations and experimental validation. Based on the synergistic use of HTL bilayer and 70-nm-thick ITO, the PeQLEDs achieved an optimal external quantum efficiency of 17.96 % at a luminance of 1763 cd/m2 and 15.19 % at 8300 cd/m2 without using any external outcoupling structure, indicating a low efficiency roll-off.

Conformal Growth of Nano‐Patterned Monolayer MoS2 with Periodic Strain via Patterned Substrate Engineering for High‐performance Photodetectors

Conformal Growth of Nano‐Patterned Monolayer MoS₂ with Periodic Strain via Patterned Substrate Engineering for High‐performance Photodetectors

Strain and Substrate-Induced Electronic Properties of Novel Mixed Anion-Based 2D ScHX2 (X = I/Br) Semiconductors.

Exploration of compounds featuring multiple anions beyond the single-oxide ion, such as oxyhalides and oxyhydrides, offers an avenue for developing materials with the prospect of novel functionality. In this paper, we present the results for a mixed anion layered material, ScHX2 (X: Br, I) based on density functional theory. The result predicted the ScHX2 (X: Br, I) monolayers to be stable and semiconducting. Notably, the electronic and mechanical properties of the ScHX2 monolayers are comparable to well-established 2D materials like graphene and MoS2, rendering them highly suitable for electronic devices. Additionally, these monolayers exhibit an ability to adjust their band gaps and band edges in response to strain and substrate engineering, thereby influencing their photocatalytic applications.

Tuning the ground state of cuprate superconducting thin films by nanofaceted substrates

Anisotropic transport properties have been assessed in a number of cuprate superconductors, providing evidence for a nematic state. We have recently shown that in ultra-thin YBa2Cu3O7−δ films, where nematicity is induced via strain engineering, there is a suppression of charge density wave scattering along the orthorhombic a-axis and a concomitant enhancement of strange metal behavior along the b-axis. Here we develop a microscopic model, that is based on the strong interaction between the substrate facets and the thin film, to account for the unconventional phenomenology. Based on the atomic force microscopy imaging of the substrates’ surface, the model is able to predict the absence (presence) of nematicity and the resulting transport properties in films grown on SrTiO3 (MgO) substrates. Our result paves the way to new tuning capabilities of the ground state of high-temperature superconductors by substrate engineering.

Influence of Substrate-Induced Charge Doping on Defect-Related Excitonic Emission in Monolayer MoS2.

Many applications of transition metal dichalcogenides (TMDs) involve transfer to functional substrates that can strongly impact their optical and electronic properties. We investigate the impact that substrate interactions have on free carrier densities and defect-related excitonic (XD) emission from MoS2 monolayers grown by metal-organic chemical vapor deposition. C-plane sapphire substrates mimic common hydroxyl-terminated substrates. We demonstrate that transferring MoS2 monolayers to pristine c-plane sapphire dramatically increases the free electron density within MoS2 layers, quenches XD emission, and accelerates exciton recombination at the optical band edge. In contrast, transferring MoS2 monolayers onto inert hexagonal boron nitride (h-BN) has no measurable influence on these properties. Our findings demonstrate the promise of utilizing substrate engineering to control charge doping interactions and to quench broad XD background emission features that can influence the purity of single photon emitters in TMDs being developed for quantum photonic applications.

Interfacial Engineering of Soft Matter Substrates by Solid-State Polymer Adsorption.

Polymer coating to substrates alters surface chemistry and imparts bulk material functionalities with a minute thickness, even in nanoscale. Specific surface modification of a substate usually requires an active substrate that, e.g., undergoes a chemical reaction with the modifying species. Here, we present a generic method for surface modification, namely, solid-state adsorption, occurring purely by entropic strive. Formed by heating above the melting point or glass transition and subsequent rinsing of the excess polymer, the emerging ultrathin (<10 nm) layers are known in fundamental polymer physics but have never been utilized as building blocks for materials and they have never been explored on soft matter substrates. We show with model surfaces as well as bulk substrates, how solid-state adsorption of common polymers, such as polystyrene and poly(lactic acid), can be applied on soft, cellulose-based substrates. Our study showcases the versatility of solid-state adsorption across various polymer/substrate systems. Specifically, we achieve proof-of-concept hydrophobization on flexible cellulosic substrates, maintaining irreversible and miniscule adsorption yet with nearly 100% coverage without compromising the bulk material properties. The method can be considered generic for all polymers whose Tg and Tm are below those of the to-be-coated adsorbed layer, and whose integrity can withstand the solvent leaching conditions. Its full potential has broad implications for diverse materials systems where surface coatings play an important role, such as packaging, foldable electronics, or membrane technology.

Enhanced interlayer electron transfer by surface treatments in mixed-dimensional van der Waals semiconductor heterostructures

We investigate the excitonic species in WS2 monolayers transferred onto III–V semiconductor substrates with different surface treatments. When the III–V substrates were covered with amorphous native oxides, negatively charged excitons dominated the spectral weight in low-temperature near-resonance photoluminescence (PL) measurements. However, when the native oxides of the III–V substrates were reduced, neutral excitons began to dominate the spectral weight, indicating a reduction in the electron density in the WS2 monolayers. The removal of the native oxides enhanced the electron transfer from the WS2 monolayer to the III–V substrate. In addition, an additional shoulder-like PL feature appeared ∼50 meV below the emission of neutral excitons, which can be attributed to the emission of localized excitons. When the III–V substrate surface was passivated by sulfur after the reduction of the native oxides, neutral excitons still dominated the spectral weight. However, the low-energy PL shoulder disappeared again, suggesting the effective delocalization of excitons through substrate surface passivation. Surface engineering of the semiconductor substrates for two-dimensional (2D) materials can provide a novel approach to control the carrier density of the 2D materials, implement deterministic carrier localization or delocalization for the 2D materials, and facilitate the interlayer transfer of charge, spin, and valley currents. These findings open the avenue for novel device concepts and phenomena in mixed-dimensional semiconductor heterostructures.

3-aminopropyltriethoxysilane modified MXene on three-dimensional nonwoven fiber substrates for low-cost, stable, and efficient solar-driven interfacial evaporation desalination

Recently, the solar-driven interfacial evaporation desalination has attracted more and more attentions due to the advantages of low cost, zero energy consumption, and high water purification rate, etc. One of the bottlenecks of this emerging technique lies in a lack of simple and low-cost ways to construct three-dimensional (3D) hierarchical microstructures for photothermal membranes. To this end, a two-step strategy is carried out by combining surface functionalization with substrate engineering. Firstly, a silane coupling agent 3-aminopropyltriethoxysilane (APTES) is grafted onto an ideal photothermal material of Ti3C2Tx MXene, to improve the nanochannel sizes and hydrophilicity, which are attributed to enlarged interspaces of MXene and introduced hydrophilic group e.g., –NH2 and –OH, respectively. Secondly, a low-cost and robust nonwoven fiber (NWF) substrate, which has a 3D micron-sized mesh structure with interlaced fiber stacks, is employed as the skeleton to load enough APTES-grafted MXene by a simple soaking method. Benefited from above design, the Ti3C2Tx-APTES/NWF composite membrane with a 3D hierarchical structure shows enhanced light scattering and utilization, water transport and vapor escape. A remarkable evaporation rate of 1.457 kg m−2 h−1 and an evaporation efficiency of 91.48 % are attained for a large-area (5 × 5 cm2) evaporator, and the evaporation rate is further increased to 1.672 kg m−2 h−1 for a small-area (2 × 2 cm2) device. The rejection rates of salt ions and heavy metal ions are higher than 99 % and 99.99 %, respectively, and the removal rates of organic dye molecules are nearly to 100 %. Besides, the composite photothermal membrane exhibits great stabilities in harsh conditions such as high salinities, long cycling, large light intensities, strong acid/alkali environments, and mechanical bending. Most importantly, the photothermal membrane shows a considerable cost-effectiveness of 89.4 g h−1/$. Hence, this study might promote the commercialization of solar-driven interfacial evaporation desalination by collaboratively considering surface modification and substrate engineering for MXene.

Synergetic substrate and additive engineering for over 30%-efficient perovskite-Si tandem solar cells

Synergetic substrate and additive engineering for over 30%-efficient perovskite-Si tandem solar cells

Unlocking the power of nano Petals: Magnifying Rhodamine 6G detection sensitivity in SERS

The escalating prevalence of hazardous, chromatic dye pollutants necessitates the implementation of a discerning and expeditious detection methodology. This investigation responds to this imperative by introducing silver-coated nickel oxide (Ag@NiO) nanostructured thin films as efficacious substrates for Surface-Enhanced Raman Spectroscopy (SERS). Interrogating the exceptional enhancement capabilities of Ag@NiO nano petals and unraveling the underlying mechanisms governing their formation, the study provides valuable insights into the optimization of SERS substrate fabrication. X-ray diffraction (XRD) analysis affirms the presence of a Face-Centered Cubic (FCC) structure within NiO, accompanied by an average crystallite size ranging from 38 to 53 nm. Scanning electron microscopy (FESEM) images unveil petal-like morphologies with widths fluctuating between 36 and 48 nm. Energy-dispersive X-ray (EDX) analysis validates the elemental composition, while UV–visible spectrum analysis delineates a band gap energy (Eg) spectrum spanning from 3.23 to 3.43 eV. Of particular significance is the showcase of Ag@NiO nano petals' astounding enhancement factor, reaching an impressive 3.2 x 107 for Rhodamine 6G (R6G). This enables detection at an extraordinary limit of 10-7 mol/L. These discernments not only propel advancements in SERS substrate engineering but also harbor implications for the meticulous analysis of R6G dye and other environmental pollutants, thereby enriching the arsenal of chemical sensing and environmental monitoring methodologies.

Darobactin Substrate Engineering and Computation Show Radical Stability Governs Ether versus C-C Bond Formation.

The Gram-negative selective antibiotic darobactin A has attracted interest owing to its intriguing fused bicyclic structure and unique targeting of the outer membrane protein BamA. Darobactin, a ribosomally synthesized and post-translationally modified peptide (RiPP), is produced by a radical S-adenosyl methionine (rSAM)-dependent enzyme (DarE) and contains one ether and one C-C cross-link. Herein, we analyze the substrate tolerance of DarE and describe an underlying catalytic principle of the enzyme. These efforts produced 51 enzymatically modified darobactin variants, revealing that DarE can install the ether and C-C cross-links independently and in different locations on the substrate. Notable variants with fused bicyclic structures were characterized, including darobactin W3Y, with a non-Trp residue at the twice-modified central position, and darobactin K5F, which displays a fused diether ring pattern. While lacking antibiotic activity, quantum mechanical modeling of darobactins W3Y and K5F aided in the elucidation of the requisite features for high-affinity BamA engagement. We also provide experimental evidence for β-oxo modification, which adds support for a proposed DarE mechanism. Based on these results, ether and C-C cross-link formation was investigated computationally, and it was determined that more stable and longer-lived aromatic Cβ radicals correlated with ether formation. Further, molecular docking and transition state structures based on high-level quantum mechanical calculations support the different indole connectivity observed for ether (Trp-C7) and C-C (Trp-C6) cross-links. Finally, mutational analysis and protein structural predictions identified substrate residues that govern engagement to DarE. Our work informs on darobactin scaffold engineering and further unveils the underlying principles of rSAM catalysis.

Enhancing the Maturation of Human Pluripotent Stem Cell-Derived Cardiomyocytes with an n-Type Organic Semiconductor Coating.

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are a promising cell source for cardiac regenerative medicine and in vitro modeling. However, hPSC-CMs exhibit immature structural and functional properties compared with adult cardiomyocytes. Various electrical, mechanical, and biochemical cues have been applied to enhance hPSC-CM maturation but with limited success. In this work, we investigated the potential application of the semiconducting polymer poly{[N,N'-bis(2-octyldodecyl)-naphthalene-1,4,5,8-bis(dicarboximide)-2,6-diyl]-alt-5,5'-(2,2'-bithiophene)} (P(NDI2OD-T2)) as a light-sensitive material to stimulate hPSC-CMs optically. Our results indicated that P(NDI2OD-T2)-mediated photostimulation caused cell damage at irradiances applied long-term above 36 μW/mm2 and did not regulate cardiac monolayer beating (after maturation) at higher intensities applied in a transient fashion. However, we discovered that the cells grown on P(NDI2OD-T2)-coated substrates showed significantly enhanced expression of cardiomyocyte maturation markers in the absence of a light exposure stimulus. A combination of techniques, such as atomic force microscopy, scanning electron microscopy, and quartz crystal microbalance with dissipation monitoring, which we applied to investigate the interface of the cell with the n-type coating, revealed that P(NDI2OD-T2) impacted the nanostructure, adsorption, and viscoelasticity of the Matrigel coating used as a cell adhesion promoter matrix. This modified cellular microenvironment promoted the expression of cardiomyocyte maturation markers related to contraction, calcium handling, metabolism, and conduction. Overall, our findings demonstrate that conjugated polymers such as P(NDI2OD-T2) can be used as passive coatings to direct stem cell fate through interfacial engineering of cell growth substrates.

Substrate Engineering toward Selective Growth of Ultrathin WC Crystals and Heterostructures via Liquid Cu‐Zn Catalyst

AbstractTransition metal carbides (TMCs) grown by chemical vapor deposition (CVD) offer promise for numerous novel phenomena and applications in the 2D limit. Despite considerable efforts thus far, the flexible customization of TMCs and their heterostructures still remains challenging. Herein, a substrate engineering is developed to achieve customized manufacturing of ultrathin WC single crystals and WC/graphene (WC‐G) heterostructures by varying the concentration of Zn in Cu‐Zn alloy substrate. It is worth noting that Zn atoms can remarkably reduce the nucleation density of graphene and promote the nucleation of WC. Thus, an increasing Zn content is applied to synergistically modulate the growth of graphene and WC, enabling the controllable fabrication of WC and WC‐G heterostructures. The synthesized WC crystals exhibit an ultrathin nature down to 3 nm, as well as high crystalline, ultra‐clean surface, and superb chemical stability. Based on that, the typical metallic properties with a temperature‐dependent resistance (nearly 1.30 Ω at 300 K and nearly 0.08 Ω at 1.7 K) and low resistance as well as excellent nonlinear optical performance of WC are demonstrated. This work provides fresh insights into regulating the growth behavior of multiblock‐structured carbides and promotes the study of their optic and electronic properties.

Regulating Spin-Valence States of Vanadium-Based Single-Atom Oxygen Reduction Catalysts through Substrate and Axial Ligand Engineering

Regulating Spin-Valence States of Vanadium-Based Single-Atom Oxygen Reduction Catalysts through Substrate and Axial Ligand Engineering

Isotopic effects on in-plane hyperbolic phonon polaritons in MoO3

Abstract Hyperbolic phonon polaritons (HPhPs), hybrids of light and lattice vibrations in polar dielectric crystals, empower nanophotonic applications by enabling the confinement and manipulation of light at the nanoscale. Molybdenum trioxide (α-MoO3) is a naturally hyperbolic material, meaning that its dielectric function deterministically controls the directional propagation of in-plane HPhPs within its reststrahlen bands. Strategies such as substrate engineering, nano- and hetero-structuring, and isotopic enrichment are being developed to alter the intrinsic dielectric functions of natural hyperbolic materials and to control the confinement and propagation of HPhPs. Since isotopic disorder can limit phonon-based processes such as HPhPs, here we synthesize isotopically enriched 92MoO3 (92Mo: 99.93 %) and 100MoO3 (100Mo: 99.01 %) crystals to tune the properties and dispersion of HPhPs with respect to natural α-MoO3, which is composed of seven stable Mo isotopes. Real-space, near-field maps measured with the photothermal induced resonance (PTIR) technique enable comparisons of in-plane HPhPs in α-MoO3 and isotopically enriched analogs within a reststrahlen band (≈820 cm−1 to ≈972 cm−1). Results show that isotopic enrichment (e.g., 92MoO3 and 100MoO3) alters the dielectric function, shifting the HPhP dispersion (HPhP angular wavenumber × thickness vs. IR frequency) by ≈−7 % and ≈+9 %, respectively, and changes the HPhP group velocities by ≈±12 %, while the lifetimes (≈3 ps) in 92MoO3 were found to be slightly improved (≈20 %). The latter improvement is attributed to a decrease in isotopic disorder. Altogether, isotopic enrichment was found to offer fine control over the properties that determine the anisotropic in-plane propagation of HPhPs in α-MoO3, which is essential to its implementation in nanophotonic applications.

Recent Advances in Spin-coating Precursor Mediated Chemical Vapor Deposition of Two-Dimensional Transition Metal Dichalcogenides.

Two-dimensional (2D) transition metal dichalcogenides (TMDs) have garnered widespread interest in the scientific community and industry for their exceptional physical and chemistry properties, and great potential for applications in diverse fields including (opto)electronics, electrocatalysis, and energy storage. Chemical vapor deposition (CVD) is one of the most compelling growth methods for the scalable growth of high-quality 2D TMDs. However, the conventional CVD process for synthesis of 2D TMDs still encounters significant challenges, primarily attributed to the high melting point of precursor powders, and achieving a uniform distribution of precursor atmosphere on the substrate to obtain controllable smaple domains is difficult. The spin-coating precursor mediated chemical vapor deposition (SCVD) strategy provides refinement over traditional methods by eliminating the use of solid precursors and ensuring a more clean and uniform distribution of the growth material on the substrate. Additionally, the SCVD process allows fine-tuning of material thickness and purity by manipulating solution composition, concentration, and the spin coating process. This Review presents a comprehensive summary of recent advances in controllable growth of 2D TMDs with a SCVD strategy. First, a series of various liquid precursors, additives, source supply methods, and substrate engineering strategies for preparing atomically thin TMDs by SCVD are introduced. Then, 2D TMDs heterostructures and novel doped TMDs fabricated through the SCVD method are discussed. Finally, the current challenges and perspectives to synthesize 2D TMDs using SCVD are discussed.

Surface‐Enhanced Raman Scattering in BIC‐Driven Semiconductor Metasurfaces

AbstractSemiconductor‐based surface‐enhanced Raman spectroscopy (SERS) substrates, as a new frontier in the field of SERS, are hindered by their poor electromagnetic field confinement and weak light‐matter interaction. Metasurfaces, a class of 2D artificial materials based on the electromagnetic design of nanophotonic resonators, enable strong electromagnetic field enhancement and optical absorption engineering for a wide range of semiconductors. However, the engineering of semiconductor substrates into metasurfaces for improving SERS activity remains underexplored. Here, an improved SERS metasurface platform is developed that leverages the combination of titanium oxide (TiO2) and the emerging physical concept of optical bound states in the continuum (BICs) to boost the Raman emission. Moreover, fine‐tuning of BIC‐assisted resonant absorption offers a pathway for maximizing the photoinduced charge transfer effect (PICT) in SERS. High values of BIC‐assisted electric field enhancement (|E/E0|2 ≈103) are achieved, challenging the preconception of weak electromagnetic (EM) field enhancement on semiconductor SERS substrates. The BIC‐assisted TiO2 metasurface platform offers a new dimension in spectrally‐tunable SERS with earth‐abundant and bio‐compatible semiconductor materials, beyond the traditional plasmonic ones.

Chemical Evolution of Enzyme-Catalyzed Glycosylation.

ConspectusThe limited availability of structurally well-defined diverse glycans remains a major obstacle for deciphering biological functions as well as biomedical applications of carbohydrates. Despite tremendous progress that has been made in past decades, the synthesis of structurally well-defined complex glycans still represents one of the most challenging topics in synthetic chemistry. Chemical synthesis of glycans is a time-consuming and labor-intensive process that requires elaborate planning and skilled personnel. In contrast, glycosyltransferase-catalyzed enzymatic synthesis provides a more efficient, convenient, low-cost, and sustainable alternative to affording diverse and complex glycans. However, the existing methods are still insufficient to fulfill the increasing demand for specific synthetic glycan libraries necessary for functional glycomics research. This is mainly attributed to the inherent character of the glycan biosynthetic pathway. In nature, there are too many glycosyltransferases involved in the in vivo glycan synthesis, but only a small number of them are available for in vitro enzymatic synthesis. For instance, humans have over 200 glycosyltransferases, but only a few of them could be produced from the conventional bacterial expression system, and most of these membrane-associated enzymes could be overexpressed only in eukaryotic cells. Moreover, the glycan biosynthetic pathway is a nontemplate-driven process, which eventually ends up with heterogeneous glycan product mixtures. Therefore, it is not a practical solution for the in vitro enzymatic synthesis of complex glycans by simply copying the glycan biosynthetic pathway.In the past decade, we have tried to develop a simplified and transformable approach to the enzymatic modular assembly of a human glycan library. Despite the structural complexity of human glycans, the glycoinformatic analysis based on the known glycan structure database and the human glycosyltransferase database indicates that there are approximately 56 disaccharide patterns present in the human glycome and only 16 disaccharide linkages are required to account for over 80% of the total disaccharide fragments, while 35 disaccharide linkages are sufficient to cover over 95% of all disaccharide fragments of human glycome. Regardless of the substrate specificity, if one glycosyltransferase could be used for the synthesis of all of the same glycosidic linkages in human glycome, it will require only a few dozen glycosyltransferases for the assembly of entire human glycans. According to the glycobioinformatics analysis results, we rationally designed about two dozen enzyme modules for the synthesis of over 20 common glycosidic linkages in human glycome, in which each enzyme module contains a glycosyltransferase and a group of enzymes for the in situ generation of a nucleotide-activated sugar donor. By sequential glycosylation using orchestrated enzyme modules, we have completed the synthesis of over 200 structurally well-defined complex human glycans including blood group antigens, O-mannosyl glycans, human milk oligosaccharides, and others. To overcome the product microheterogeneity problem of enzymatic synthesis in the nontemplate-driven glycan biosynthetic pathway, we developed several substrate engineering strategies to control or manipulate the outcome of glycosyltransferase-catalyzed reactions for the precise synthesis of structurally well-defined isomeric complex glycans.