Essential Active Site Research Articles

Atomic-Layer IrOx Enabling Ligand Effect Boosts Water Oxidation Electrocatalysis.

An in situ formed IrOx (x ≤ 2) layer driven by anodic bias serves as the essential active site of Ir-based materials for oxygen evolution reaction (OER) electrocatalysis. Once being confined to atomic thickness, such an IrOx layer possesses both a favorable ligand effect and maximized active Ir sites with a lower O-coordination number. However, limited by a poor understanding of surface reconstruction dynamics, obtaining atomic layers of IrOx remains experimentally challenging. Herein, we report an idea of material design using intermetallic IrVMn nanoparticles to induce in situ formation of an ultrathin IrOx layer (O-IrVMn/IrOx) to enable the ligand effect for achieving superior OER electrocatalysis. Theoretical calculations predict that a strong electronic interaction originating from an orderly atomic arrangement can effectively hamper the excessive leaching of transition metals, minimizing vacancies for oxygen coordination. Linear X-ray absorption near edge spectra analysis, extended X-ray absorption fine structure fitting outcomes, and X-ray photoelectron spectroscopy collectively confirm that Ir is present in lower oxidation states in O-IrVMn/IrOx due to the presence of unsaturated O-coordination. Consequently, the O-IrVMn/IrOx delivers excellent acidic OER performances with an overpotential of only 279 mV at 10 mA cm-2 and a high mass activity of 2.3 A mg-1 at 1.53 V (vs RHE), exceeding most Ir-based catalysts reported. Moreover, O-IrVMn/IrOx also showed excellent catalytic stability with only 0.05 at. % Ir dissolution under electrochemical oxidation, much lower than that of disordered D-IrVMn/IrOx (0.20 at. %). Density functional theory calculations unravel that the intensified ligand effect optimizes the adsorption energies of multiple intermediates involved in the OER and stabilizes the as-formed catalytic IrOx layer.

Unlocking plateau capacity with versatile precursor crosslinking for carbon anodes in Na-ion batteries

As the precursor material inherently determines the fundamental structure of hard carbons, a direct manipulation of precursors at the molecular level promises enhanced flexibility in designing hard carbon architectures, which plays a pivotal role in dictating the final microstructural properties and, ultimately, the overall sodium storage performance. In this study, we present a novel generalized strategy utilizing P and O double cross-linking to convert pitch into a thermosetting precursor, creating copious micropores within pitch-based carbon. These micropores serve as essential pathways and active binding sites for sodium ion transport and storage, leading to pitch-derived hard carbons with a remarkable specific capacity of 416.1 mAh/g and an impressive initial Coulombic efficiency of 89.7%. Extensive study revealed a strong correlation between the increased plateau capacity and the closed pore volume, validating the micropore-driven sodium ion storage mechanism. Our findings underscores the groundbreaking significance of cross-linking in precursor modification, paving the way for the design and synthesis of high-performance hard carbon anodes for next-generation Na-ion batteries.

Whole genome identification, molecular docking and expression analysis of enzymes involved in the selenomethionine cycle in Cardamine hupingshanensis

BackgroundThe selenomethionine cycle (SeMTC) is a crucial pathway for the metabolism of selenium. The basic bioinformatics and functions of four enzymes involved in the cycle including S-adenosyl-methionine synthase (MAT), SAM-dependent methyltransferase (MTase), S-adenosyl-homocysteine hydrolase (SAHH) and methionine synthase (MTR), have been extensively reported in many eukaryotes. The identification and functional analyses of SeMTC genes/proteins in Cardamine hupingshanensis and their response to selenium stress have not yet been reported.ResultsIn this study, 45 genes involved in SeMTC were identified in the C. hupingshanensis genome. Phylogenetic analysis showed that seven genes from ChMAT were clustered into four branches, twenty-seven genes from ChCOMT were clustered into two branches, four genes from ChSAHH were clustered into two branches, and seven genes from ChMTR were clustered into three branches. These genes were resided on 16 chromosomes. Gene structure and homologous protein modeling analysis illustrated that proteins in the same family are relatively conserved and have similar functions. Molecular docking showed that the affinity of SeMTC enzymes for selenium metabolites was higher than that for sulfur metabolites. The key active site residues identified for ChMAT were Ala269 and Lys273, while Leu221/231 and Gly207/249 were determined as the crucial residues for ChCOMT. For ChSAHH, the essential active site residues were found to be Asn87, Asp139 and Thr206/207/208/325. Ile204, Ser111/329/377, Asp70/206/254, and His329/332/380 were identified as the critical active site residues for ChMTR. In addition, the results of the expression levels of four enzymes under selenium stress revealed that ChMAT3-1 genes were upregulated approximately 18-fold, ChCOMT9-1 was upregulated approximately 38.7-fold, ChSAHH1-2 was upregulated approximately 11.6-fold, and ChMTR3-2 genes were upregulated approximately 28-fold. These verified that SeMTC enzymes were involved in response to selenium stress to varying degrees.ConclusionsThe results of this research are instrumental for further functional investigation of SeMTC in C. hupingshanensis. This also lays a solid foundation for deeper investigations into the physiological and biochemical mechanisms underlying selenium metabolism in plants.

Phosphazene-Based Covalent Organic Framework as an Efficient Catalyst (COF-1) for the Dehydration of Fructose to 5-HMF.

5-Hydroxymethylfurfural (HMF) is a promising organic platform for producing value-added chemicals. In this work, we focused on using a covalent organic framework (COF-1) as a heterogeneous catalyst for the dehydration of fructose to 5-HMF. The unique phosphazene unit-functionalized pores of COF-1 are essential active sites for catalytic performance. The results show that under the optimized reaction conditions, a maximum yield of 90% was obtained within 1.5 h at 120 °C. Furthermore, the effects of the catalyst load, reaction temperature, and usage of solvents for the improvement of reaction yield were investigated. The catalyst recyclability results showed that the yield of HMF did not change appreciably (90-82%) over five consecutive recycling runs. This work provides a viable strategy by applying phosphazene-based COF-1 for the efficient synthesis of HMF from renewable biomass. The synthesized HMF was further used for the synthesis of the biopolymer monomer furan-2,5-dimethylcarboxylate (FDMC) through N-heterocyclic carbene (NHC)-catalyzed oxidative esterification.

Mechanistic plasticity in ApmA enables aminoglycoside promiscuity for resistance.

The efficacy of aminoglycoside antibiotics is waning due to the acquisition of diverse resistance mechanisms by bacteria. Among the most prevalent are aminoglycoside acetyltransferases (AACs) that inactivate the antibiotics through acetyl coenzyme A-mediated modification. Most AACs are members of the GCN5 superfamily of acyltransferases which lack conserved active site residues that participate in catalysis. ApmA is the first reported AAC belonging to the left-handed β-helix superfamily. These enzymes are characterized by an essential active site histidine that acts as an active site base. Here we show that ApmA confers broad-spectrum aminoglycoside resistance with a molecular mechanism that diverges from other detoxifying left-handed β-helix superfamily enzymes and canonical GCN5 AACs. We find that the active site histidine plays different functions depending on the acetyl-accepting aminoglycoside substrate. This flexibility in the mechanism of a single enzyme underscores the plasticity of antibiotic resistance elements to co-opt protein catalysts in the evolution of drug detoxification.

Identifying the active sites in unequal iron-nitrogen single-atom catalysts

Single-atom catalysts (SACs) have become one of the most attractive frontier research fields in catalysis and energy conversion. However, due to the atomic heterogeneity of SACs and limitations of ensemble-averaged measurements, the essential active sites responsible for governing specific catalytic properties and mechanisms remain largely concealed. In this study, we develop a quantitative method of single-atom catalysis–fluorescence correlation spectroscopy (SAC-FCS), leveraging the atomic structure-dependent catalysis kinetics and single-turnover resolution of single-molecule fluorescence microscopy. This method enables us to investigate the oxidase-like single-molecule catalysis on unidentical iron-nitrogen (Fe-N) coordinated SACs, quantifying the active sites and their kinetic parameters. The findings reveal the significant differences of single sites from the average behaviors and corroborate the oxidase-like catalytic mechanism of the Fe-N active sites. We anticipate that the method will give essential insights into the rational design and application of SACs.

Mapping the Initial Stages of a Protective Pathway that Enhances Catalytic Turnover by a Lytic Polysaccharide Monooxygenase.

Oxygenase and peroxygenase enzymes generate intermediates at their active sites which bring about the controlled functionalization of inert C-H bonds in substrates, such as in the enzymatic conversion of methane to methanol. To be viable catalysts, however, these enzymes must also prevent oxidative damage to essential active site residues, which can occur during both coupled and uncoupled turnover. Herein, we use a combination of stopped-flow spectroscopy, targeted mutagenesis, TD-DFT calculations, high-energy resolution fluorescence detection X-ray absorption spectroscopy, and electron paramagnetic resonance spectroscopy to study two transient intermediates that together form a protective pathway built into the active sites of copper-dependent lytic polysaccharide monooxygenases (LPMOs). First, a transient high-valent species is generated at the copper histidine brace active site following treatment of the LPMO with either hydrogen peroxide or peroxyacids in the absence of substrate. This intermediate, which we propose to be a CuII-(histidyl radical), then reacts with a nearby tyrosine residue in an intersystem-crossing reaction to give a ferromagnetically coupled (S = 1) CuII-tyrosyl radical pair, thereby restoring the histidine brace active site to its resting state and allowing it to re-enter the catalytic cycle through reduction. This process gives the enzyme the capacity to minimize damage to the active site histidine residues "on the fly" to increase the total turnover number prior to enzyme deactivation, highlighting how oxidative enzymes are evolved to protect themselves from deleterious side reactions during uncoupled turnover.

Ethanol dehydrogenation to acetaldehyde over a Cuδ+-based Cu-MFI catalyst

Copper catalysts have been extensively investigated for the dehydrogenation of ethanol to acetaldehyde. However, identifying the essential active sites for this reaction is difficult because of the complex coordination structure and variable valences of the Cu species during the reaction. The stability of the Cu catalysts in the reaction also needs to be substantially improved. In this study, Cu-MFI, a well-defined Cu-based Lewis acid catalyst, was prepared using a post-acid treatment method for ethanol dehydrogenation. Different from the widely reported Cu+ and Cu0 species accounting for the activity of Cu catalysts, conditional experiments and in situ characterizations revealed that the highly dispersed Cuδ+ (1 <δ < 2) species on the MFI support are the essential active sites for ethanol dehydrogenation. Due to the strong interaction between Cu and silica via the Cu–O–Si linkage, the Cuδ+ species were very stable in the reaction and played the role of a Lewis acid catalyst in promoting ethanol activation and dehydrogenation. Over the optimal catalyst 5%Cu-MFI-deCu, 95% selectivity of acetaldehyde and approximately 87% ethanol conversion were obtained at 250 °C and a weight hourly space velocity of 0.64 h−1 in 120 h time on stream.

7-Deazaguanines in DNA: functional and structural elucidation of a DNA modification system.

The modified nucleosides 2'-deoxy-7-cyano- and 2'-deoxy-7-amido-7-deazaguanosine (dPreQ0 and dADG, respectively) recently discovered in DNA are the products of the bacterial queuosine tRNA modification pathway and the dpd gene cluster, the latter of which encodes proteins that comprise the elaborate Dpd restriction-modification system present in diverse bacteria. Recent genetic studies implicated the dpdA, dpdB and dpdC genes as encoding proteins necessary for DNA modification, with dpdD-dpdK contributing to the restriction phenotype. Here we report the in vitro reconstitution of the Dpd modification machinery from Salmonella enterica serovar Montevideo, the elucidation of the roles of each protein and the X-ray crystal structure of DpdA supported by small-angle X-ray scattering analysis of DpdA and DpdB, the former bound to DNA. While the homology of DpdA with the tRNA-dependent tRNA-guanine transglycosylase enzymes (TGT) in the queuosine pathway suggested a similar transglycosylase activity responsible for the exchange of a guanine base in the DNA for 7-cyano-7-deazaguanine (preQ0), we demonstrate an unexpected ATPase activity in DpdB necessary for insertion of preQ0 into DNA, and identify several catalytically essential active site residues in DpdA involved in the transglycosylation reaction. Further, we identify a modification site for DpdA activity and demonstrate that DpdC functions independently of DpdA/B in converting preQ0-modified DNA to ADG-modified DNA.

The ChaC1 active site: Defining the residues and determining the role of ChaC1-exclusive residues in the structural and functional stability.

The glutathione degrading enzyme ChaC1 is highly upregulated in several cancers and viral infections making it a potential pharmacological target for cancer therapy. As an enzyme, however, ChaC1 has a relatively high Km (~2 mM) towards its natural substrate, and therefore finding its inhibitors becomes very difficult. Given this limitation, a careful mapping of the active site has become necessary. In the current study, the enzyme-substrate complex was generated by docking glutathione with the modeled hChaC1 structure. Using a combination of in silico and wet lab approaches, the active site residues forming direct interactions with the substrate glutathione were identified and validated. Furthermore, the role of residues exclusively conserved in the ChaC family and forming the surface of the active site were also explored for their putative role in active site stabilization. Mutants of these residues have been analysed for their structural stability and interaction with the substrate through MD simulations and MMGBSA binding energy calculations. These findings were experimentally validated by assessment of their function through in vivo assays in yeast. The experimental evidences along with the molecular modeling suggest that residues 38'YGSL'41, D68, R72, E115, and Y143 are responsible for high affinity binding of hChaC1 with the substrate/inhibitor, whereas the residues exclusive to the ChaC family are required for the structural stability of the enzyme and its active site. Such a characterization of essential active site and conserved residues is significant as a key step toward rational design of novel inhibitors of the ChaC1 enzyme.

Effective peroxymonosulfate activation using electrified nanohybrid filter towards one-step decontamination of roxarsone: Performance and mechanism

An electrochemical filtration technology was rationally developed toward effective one-step roxarsone (ROX) decontamination. A bifunctional electroactive nanohybrid filter of carbon nanotube (CNT) and iron(III) oxide (Fe3O4) species is the key to this technology. ROX (91.8%) was simultaneously adsorbed and oxidized by passing through a functional filter together with peroxymonosulfate (PMS) under an electric field (τ < 2 s). Several critical operational parameters of the system performance (applied voltage, PMS concentration, solution pH and aqueous competitive anions) were identified and optimized. The underlying mechanism was proposed based on the experimental evidence. Mechanistic studies have demonstrated that carbonyl group (CO) of CNT can serve as electron donor to activate PMS to generate singlet oxygen (1O2) and Fe can provide electrons to PMS to produce hydroxyl radical (HO•) and sulfate radical (SO4•–). This suggests that both radicals and non-radical pathways contribute to the effective oxidation of ROX during the electrochemical filtration process. These reactions can transform highly toxic arsenite (As(III)) to less toxic arsenate (As(V)) by cleavage of the arsenic–carbon bond (As–C). The applied electric field facilitated the cycling of the ferric/ferrous (Fe3+/Fe2+) pairs and induced the electro-PMS activation to generate aggressive radical species. Meanwhile, the carbonyl groups of CNT provided essential active sites for the generation of 1O2. The synergistic interplays of the electrochemical reactivity, high porosity, flow-through configuration and abundant active sites for effective ROX decontamination were highlighted. The findings of this study are dedicated to providing a viable solution to remove organic arsenic and related species from water.

Flow-through electrochemical removal of benzotriazole by electroactive ceramic membrane

Benzotriazole (BTA) is a widely used anticorrosive additive that is of endurance, bioaccumulation and toxicity, and BTA industrial wastewater treatment remains a challenge. This study reports efficient electrochemical removal of BTA by titanium oxide (TiSO) electroactive ceramic membrane (ECM), indicated by 98.1% removal at current density of 20 mA∙cm−2 and permeate flux of 692 LHM under cathode-to-anode flow pattern (1 h). Electrochemical analysis demonstrated the pH-dependent formation of anti-corrosive BTA film on the TiSO anode, which was responsible for improved BTA removal for cathode-to-anode (CA) flow pattern compared with that for anode-to-cathode (AC). The modelling results showed the CA flow pattern to be more favourable for BTA oxidation mediated by electro-generated •OH by preventing the formation of deactivation film via creating an alkaline boundary layer at the anode/electrolyte interface. Intermediates and essential active sites were identified by using experimental analysis and theoretical density functional theory (DFT) calculations, thereby the most likely degradation pathways were underlined. Toxicity analysis revealed remarkable decrease in oral rat LD50 values and bioaccumulation factor during electrochemical degradation of BTA. This study provides a proof-in-concept demonstration of effective removal for anti-corrosive emerging pollutants by TiSO-ECM under flow-through pattern.

Experimental and theoretical studies of cadmium adsorption over Fe2O3 sorbent in incineration flue gas

Iron oxide (Fe2O3) has been used as an effective sorbent to remove various heavy metals due to its large adsorption capacity and low cost. The cadmium capture capability of Fe2O3 sorbent was investigated in the simulated flue gas at the temperatures of 700–1100 °C. Fe2O3 sorbent exhibited much better Cd adsorption capability than SiO2 and Al2O3 sorbents. The largest Cd adsorption capacity of Fe2O3 sorbent was 86.7 mg/g at 700 °C. The XPS analysis demonstrated that Fe3+ might partially participate in the cadmium adsorption process. A large quantity of CdO was generated and adsorbed on the Fe2O3 surface. The density functional theory (DFT) calculations were used to determine the active sites and the involved mechanism of different cadmium species adsorption over Fe2O3 sorbent. Cd, CdCl2, and CdO can be stably adsorbed on the Fe2O3 surface with the adsorption energies of −40.64 kJ/mol, −237.42 kJ/mol, and −375.03 kJ/mol, respectively. The results illustrate that the Fe atoms are the essential active sites for cadmium species adsorption on Fe2O3 surface. The orbital hybridization between cadmium atom and Fe site suggests that the strong interaction occurs during Cd adsorption process. Both physisorption and chemisorption mechanisms were responsible for the cadmium capture by Fe2O3 sorbent.

Seed specifically over-expressing DGAT2A enhances oil and linoleic acid contents in soybean seeds

Triacylglycerol (TAG), a main component of oil, is mainly biosynthesized by diacylglycerol acyltransferase (DGAT), which is critical for oil accumulation in plants. Intensive focus has been on DGAT2 functioning in unsaturated fatty acids biosynthesis. In this study, we analyzed the coding sequence (CDS) and amino acid sequence of GmDGAT2A and determined its key active sites through site-directed mutagenesis. As a consequence, H132, G201, and P152-X-I154-K155 were found to be essential active sites for GmDGAT2A. The spatial structure of the protein may bring the three active sites into close proximity, constituting an active domain. Additionally, N-terminus of GmDGAT2A was found to be an important regulator for the activity. Further, in vitro activity results uncovered GmDGAT2A was prone to utilize C18:2-CoA as the substrate. Consequently, overexpression of GmDGAT2A driven by a seed-specific promoter of Gmole1 in soybean significantly increased linoleic acid content specifically and total oil content, concomitant with accelerated elongation.

Structure of 3'-PO4/5'-OH RNA ligase RtcB in complex with a 5'-OH oligonucleotide.

RtcB enzymes comprise a widely distributed family of manganese- and GTP-dependent RNA repair enzymes that join 2′,3′-cyclic phosphate ends to 5′-OH ends via RtcB-(histidinyl-N)–GMP, RNA 3′-phosphate, and RNA3′pp5′G intermediates. RtcB can ligate either 5′-OH RNA or 5′-OH DNA strands in vitro. The nucleic acid contacts of RtcB are uncharted. Here we report a 2.7 Å crystal structure of Pyrococcus horikoshii RtcB in complex with a 6-mer 5′-OH DNA oligonucleotide HOA1pT2pG3pT4pC5pC6, which reveals enzymic contacts of Asn202 to the terminal 5′-OH nucleophile; Arg238 to the A1pT2 and T2pG3 phosphates; Arg190 and Gln194 to the T2pG3 phosphate; and an Arg190 π-cation interaction with the G3 nucleobase. The structural insights affirm functional studies of E. coli RtcB that implicated the conserved counterpart of Arg238 in engagement of the 5′-OH strand for ligation. The essential active site Cys98 that coordinates two manganese ions is oxidized to cysteine sulfonic acid in our structure, raising the prospect that RtcB activity might be sensitive to modulation during oxidative stress.

Alkane isomerization on highly reduced Pd/Al2O3 catalysts. The crucial role of Pd-Al species

Highly reduced (at 500 and 600 °C) 1 wt% Pd/Al2O3 samples showed remarkable isomerization selectivity in n-hexane conversion, >80%, in agreement with previous reports. The catalytic behavior of highly reduced Pd/Al2O3 becomes typical of platinum, the metal regarded as unique in alkane isomerization. Although not detected in low metal loaded catalysts, Pd-Al species evidenced by formation of Pd-Al alloys in 5 wt% metal loaded catalysts appear essential active sites responsible for n-hexane isomerization. Other possible reasons (contribution from alumina acidity and changes in the structure of metal surface) were analyzed and discarded. A mechanistic rationale for the improvement of n-hexane isomerization is outlined.

Insights into a dual function amide oxidase/macrocyclase from lankacidin biosynthesis

Acquisition of new catalytic activity is a relatively rare evolutionary event. A striking example appears in the pathway to the antibiotic lankacidin, as a monoamine oxidase (MAO) family member, LkcE, catalyzes both an unusual amide oxidation, and a subsequent intramolecular Mannich reaction to form the polyketide macrocycle. We report evidence here for the molecular basis for this dual activity. The reaction sequence involves several essential active site residues and a conformational change likely comprising an interdomain hinge movement. These features, which have not previously been described in the MAO family, both depend on a unique dimerization mode relative to all structurally characterized members. Taken together, these data add weight to the idea that designing new multifunctional enzymes may require changes in both architecture and catalytic machinery. Encouragingly, however, our data also show LkcE to bind alternative substrates, supporting its potential utility as a general cyclization catalyst in synthetic biology.

Highly Sensitive Lysine Deacetylase Assay Based on Acetylated Firefly Luciferase

Lysine deacetylases (KDACs) play important roles in many physiological processes and are implicated in many human diseases. Hence, the search for modulators of KDACs is very active, and reliable assays for monitoring their activity are key to success. Here, we describe a new KDAC assay based on Firefly luciferase harboring an acetylation on an essential active site lysine. We show that several KDACs can reverse this modification and hence activate luciferase. This new assay is extremely sensitive, reliable, and fast and can be performed in a continuous format. We used this assay to screen a small library of compounds and identified several novel effectors of SirT2 with low micromolar activity.

Gene Cloning, Prokaryotic Expression, and Biochemical Characterization of a Soluble Trehalase in Helicoverpa armigera Hübner (Lepidoptera: Noctuidae).

Trehalase is an indispensable component of insect hemolymph that plays important role in energy metabolism and stress resistance. In this study, we cloned and expressed the gene encoding soluble trehalase (HaTreh-1) of Helicoverpa armigera (cotton bollworm) and characterized the enzyme. HaTreh-1 had a full-length open reading frame encoding a protein of 571 amino acids. Sequence comparison indicated that HaTreh-1 was similar to some known insect trehalases. Two essential active sites (D321 and E519) and three essential residues (R168, R221, and R286) were conserved in HaTreh-1. The recombinant trehalase was expressed in Escherichia coli and purified by nickel exchange chromatography. Molecular weight of the recombinant protein was about 71 kDa, and the optimum HaTreh-1 enzyme activity is at 55°C with pH 6.0. Enzymatic assays showed a Km value of 72.8 mmol/liter and a Vmax value of 0.608 mmol/(liter·min). Inhibition assays in vitro indicated that castanospermine, a polyhydroxylated alkaloid, was an effective competitive inhibitor of trehalase with a Ki value of 6.7 μmol/liter. The inhibitor action of castanospermine was linked to its modification effect on trehalase structure. The circular dichroism spectrum showed that the percentage of α-helix increased under the presence of castanospermine. Results of our study will aid in developing effective trehalase inhibitors for controlling H. armigera in the future.

Ultrafast response and recovery trimethylamine sensor based on α-Fe2O3 snowflake-like hierarchical architectures

Snowflake-like α-Fe2O3 hierarchical architectures have been developed to detect trimethylamine (TMA) gas. The α-Fe2O3 has been synthesized by using a facile solvothermal method and a subsequent annealing process. As-fabricated snowflake-like α-Fe2O3 based sensor exhibits ultrafast response and recovery time (0.9 s/1.5 s), excellent selectivity, high response, and superior repeatability toward TMA gas. XPS, PL and UV indicates that abundant oxygen vacancies exist in the α-Fe2O3 surfaces. Oxygen vacancies provide essential active sites for gas adsorption which enormously improve the ability of electron transfer. The remarkable TMA sensing performance can be attributed to the cooperation of oxygen vacancies providing more active sites, rapid electron exchange between Fe3+ and Fe2+ cations inducing conductivity greatly change, and unique snowflake-like hierarchical structure possessing large contacting area and monocrystalline feature. Such novel snowflake-like α-Fe2O3 hierarchical architectures will be a promising candidate for TMA, fish and sea-food freshness detection.