Molecular Modification Research Articles

A novel 3D-QSAR model assisted by coefficient of variation method and its application in FQs’ modification

Methods of the root-mean-square normalization and the coefficient of variation weighting were employed to normalize and weight the biodegradation effect values of aerobic bacteria and fungi of FQs molecules, and then the comprehensive biodegradation effect 3D-QSAR model of FQs was constructed using the comprehensive biodegradation effect value as target function. It was obtained from the three-dimensional equipotential map of the constructed 3D-QSAR model, the substitution sites and substitution groups that affect the comprehensive biodegradability of FQs were screened, and the widely used levofloxacin (named FQ-16) as target molecule was selected and modified with total of 67 FQ-16 derivatives designed. It was also found that there were 5 kinds of FQ-16 derivatives’ aerobic bacteria and fungi biodegradability ratio, which were predicted using FQs’ aerobic bacteria and fungi biodegradability 3D QSARs, approaching to the weight ratio set by the coefficient of variation method (55.47:44.53), indicating the comprehensive biodegradation effect 3D-QSAR model of FQs effectively covering both aerobic bacteria and fungi biodegradation information, and three of above FQ-16 derivatives also passed through the functionality and photodegradability assessment with significantly improvement. Change rate in energy barrier of FQ-16 biodegradation before and after modification agreed well with the increase in their comprehensive biodegradation effect values, meaning that the designed FQ-16 derivatives’ biodegradation was confirmed by biodegradation pathway simulation. This article aims to provide a theoretical method for constructing FQs molecular multi-effect 3D-QSAR model and its environment-friendly molecular modification.

Molecular Modification of Single Cobalt Sites Boosts the Catalytic Activity of CO2 Electroreduction into CO

Molecular Modification of Single Cobalt Sites Boosts the Catalytic Activity of CO₂ Electroreduction into CO

Association between NPPA promoter methylation and hypertension: results from Gusu cohort and replication in an independent sample

BackgroundAtrial natriuretic peptide (ANP), one of the main members of the natriuretic peptides system, has been associated with hypertension and related complications, but the underlying molecular mechanisms are not very clear. Here, we aimed to examine whether DNA methylation, a molecular modification to the genome, of the natriuretic peptide A gene (NPPA), the coding gene of ANP, was associated with hypertension.MethodsPeripheral blood DNA methylation of NPPA promoter was quantified by target bisulfite sequencing in 2498 community members (mean aged 53 years, 38% men) as a discovery sample and 1771 independent participants (mean aged 62 years, 54% men) as a replication sample. In both samples, we conducted a single CpG association analysis, followed by a gene-based association analysis, to examine the association between NPPA promoter methylation and hypertension, adjusting for age, sex, education level, cigarette smoking, alcohol consumption, obesity, fasting glucose, and lipids. Multiple testing was controlled by the false discovery rate approach.ResultsOf the 9 CpG loci assayed, hypermethylation at 5 CpGs (CpG1, CpG3, CpG6, CpG8, and CpG9) was significantly associated with a lower odds of prevalent hypertension in the discovery sample, and one CpG methylation (CpG1 located at Chr1:11908353) was successfully replicated in the replication sample (OR = 0.82, 95%CI 0.74–0.91, q = 0.002) after adjusting for covariates and multiple testing. The gene-based analysis found that DNA methylation of the 9 CpGs at NPPA promoter as a whole was significantly associated with blood pressure and prevalent hypertension in both samples (all P < 0.05).ConclusionsDNA methylation levels at NPPA promoter were decreased in Chinese adults with hypertension. Aberrant DNA methylation of the NPPA gene may participate in the mechanisms of hypertension.

Associations of B-Type Natriuretic Peptide and Its Coding Gene Promoter Methylation With Functional Outcome of Acute Ischemic Stroke: A Mediation Analysis.

BackgroundThe prognostic role of B‐type natriuretic peptide (BNP) in stroke has been suggested, but limited studies have shown mixed results and unknown underlying mechanisms. DNA methylation, a molecular modification that alters gene expression, may represent a candidate mechanism for this purpose. We aimed to examine the associations of BNP and methylation of its coding gene (natriuretic peptide B [NPPB]) with the functional outcome in a large sample of patients with acute ischemic stroke from CATIS (China Antihypertensive Trial in Acute Ischemic Stroke).Methods and ResultsLeveraging participants from CATIS with available specimens, serum proBNP (equimolarly produced with BNP) was measured in 3216 patients (mean age, 62 years; 64% men), and peripheral blood DNA methylation of the NPPB promoter was quantified by targeted bisulfite sequencing in 806 patients (mean age, 62 years; 54% men). The functional outcome was defined as an ordered modified Rankin Scale score assessed at 14 days or hospital discharge after stroke onset. Mediation analysis was conducted to test the potential mediating effect of proBNP on the relationship between NPPB methylation and functional outcome. The results showed that a higher level of proBNP was significantly associated with a higher risk of having a poorer functional outcome (odds ratio [OR], 1.14; P=0.006). Every 5% of hypermethylation at 2 (Chr1:11919160 [OR, 0.93; P=0.022] and Chr1:11918989 [OR, 0.92; P=0.032]) of 11 CpG loci assayed was associated with 7% and 8% lower risk, respectively, of having a poor functional outcome. In addition, proBNP was negatively correlated to hypermethylation at 1 CpG (Chr1:11918989 [β=−0.029; P=0.009]) and mediated approximately 7.69% (95% CI, 2.50%–13.82%) of the association between this CpG methylation and the functional outcome.ConclusionsHypermethylation at the NPPB promoter is associated with the functional outcome after ischemic stroke, at least partially by suppressing BNP expression or excretion.

Phosphonic Acid Modified ZnO Nanowire Sensors: Directing Reaction Pathway of Volatile Carbonyl Compounds.

Surface molecular transformations on nanoscale metal oxides are inherently complex, and directing those reaction pathways is still challenging but important for designing their various applications, including molecular sensing, catalysts, and others. Here, a rational strategy to direct a reaction pathway of volatile carbonyl compounds (nonanal: biomarker) on single-crystalline ZnO nanowire surfaces via molecular modification is demonstrated. The introduction of a methylphosphonic acid modification on the ZnO nanowire surface significantly alters the surface reaction pathway of nonanal via suppressing the detrimental aldol condensation reaction. This is directed by intentionally decreasing the probability of two neighboring molecular activations on the nanowire surface. Spectrometric measurements reveal the correlation between the suppression of the aldol condensation surface reaction and the improvement in the sensor performance. This tailored surface reaction pathway effectively reduces the operating temperature from 200 to 100 °C while maintaining the sensitivity. This is because the aldol condensation product ((E)-2-heptyl-2-undecenal) requires a higher temperature to desorb from the surface. Thus, the proposed facile strategy offers an interesting approach not only for the rational design of metal oxide sensors for numerous volatile carbonyl compounds but also for tailoring various surface reaction pathways on complex nanoscale metal oxides.

Reaction Rate Governs the Viscoelasticity and Nanostructure of Folded Protein Hydrogels.

Hydrogels constructed from folded protein domains are of increasing interest as resilient and responsive biomaterials, but their optimization for applications requires time-consuming and costly molecular design. Here, we explore a complementary approach to control their properties by examining the influence of crosslinking rate on the structure and viscoelastic response of a model hydrogel constructed from photochemically crosslinked bovine serum albumin (BSA). Gelation is observed to follow a heterogeneous nucleation pathway in which BSA monomers crosslink into compact nuclei that grow into fractal percolated networks. Both the viscoelastic response probed by shear rheology and the nanostructure probed by small-angle X-ray scattering (SAXS) are shown to depend on the photochemical crosslinking reaction rate, with increased reaction rates corresponding to higher viscoelastic moduli, lower fractal dimension, and higher fractal cluster size. Reaction rate-dependent changes are shown to be consistent with a transition between diffusion- and rate-limited assembly, and the corresponding changes to viscoelastic response are proposed to arise from the presence of nonfractal depletion regions, as confirmed by SAXS. This controllable nanostructure and viscoelasticity constitute a potential route for the precise control of hydrogel properties, without the need for molecular modification.

Molecular Interaction, Chain Conformation, and Rheological Modification during Electrospinning of Hyaluronic Acid Aqueous Solution.

Most of natural water-soluble polymers are difficult to electrospin due to their specific chain conformation in aqueous solution, which limits their applications. This study investigated the effects of polyethylene oxide (PEO) on the electrospinning of hyaluronic acid (HA) in HA/PEO aqueous solutions. The rheological properties of HA/PEO aqueous solutions showed polymer chain entanglement in HA was the essential factor affecting its electrospinnability. Wide-angle X-ray scattering and differential scanning calorimetry analyses of a PEO crystal showed different crystallization behavior of the PEO chain with different molecular weight, which indicates different interaction with HA. A schematic molecular model has been proposed to explain the effect of PEO on the chain conformation of HA along with the relationship between electrospinnability and chain entanglement. PEO with a relatively high molecular weight with limited crystal formation formed extensive chain entanglements with HA, while PEO with relatively low molecular weight weakened the interactions among HA chains. The findings of this study provide a wide perspective to better understand the electrospinning mechanisms of natural polyelectrolytes and usage in tissue engineering.

A Modified 3D-QSAR Model Based on Ideal Point Method and Its Application in the Molecular Modification of Plasticizers with Flame Retardancy and Eco-Friendliness.

The addition of plasticizers makes plastics flammable, and thus, poses a potential risk to the environment. In previous researches, plasticizers with flame retardancy had been synthesized, but their eco-friendliness had not been tested or described. Thus, in this paper, eco-friendliness plasticizers with flame retardancy were designed based on phthalic acid esters (PAEs), which are known as common plasticizers and major plastic additives. For a comprehensive analysis, such as flammability, biotoxicity, and enrichment effects, 17 PAEs’ comprehensive evaluation values were calculated based on the ideal point method. Further, a multi-effect three-dimensional quantitative structure-activity relationship (3D-QSAR) model of PAEs’ flammability, biotoxicity and enrichment effects was constructed. Thus, 18 dimethyl phthalate (DMP) derivatives and 20 diallyl phthalate (DAP) derivatives were designed based on three-dimensional contour maps. Through evaluation of eco-friendliness and flammability, six eco-friendly PAE derivatives with flame retardancy were screened out. Based on contour maps analysis, it was confirmed that the introduction of large groups and hydrophobic groups was beneficial to the simultaneous improvement of PAEs’ comprehensive effects, and multiple effects. In addition, the group properties were correlated significantly with improved degrees of the comprehensive effects of corresponding PAE derivatives, confirming the feasibility of the comprehensive evaluation method and modified scheme.

Molecular structural modification of duck egg white protein conjugates with monosaccharides for improving emulsifying capacity

Glycosylation is considered to be an effective method to improve the emulsifying property of proteins, and this work investigates and discusses the correlation between the molecular structure of glycosylated egg white protein (EWP) and its emulsifying capacity. Three types of monosaccharides, including glucose, d-galactose and d-xylose, were utilized to conjugate with EWP in a wet glycosylation environment. Molecular structural properties of EWP, involving surface hydrophobicity, secondary and tertiary structures, protein flexibility, and free sulfhydryl group content, were determined. The results showed that the degree of grafting (DG), browning intensity and apparent viscosity increased significantly (P < 0.05) with the monosaccharides proportion improved, and the intrinsic fluorescence intensity and surface hydrophobicity of EWP decreased obviously during glycosylation, and the absolute value of zeta potential, protein flexibility and free sulfhydryl content improved remarkably (P < 0.05). Emulsifying capacity results showed that glycosylation improved emulsifying activity and emulsifying stability of EWP, especially after the addition of d-xylose. The particle size analysis exhibited that the D4,3 and D3,2 declined obviously (P < 0.05) as DG increased. Correlation analysis revealed the improvement of the emulsifying capacity of glycosylated EWP showed in a DG-dependent manner, as remarkably influenced by molecular flexibility, surface hydrophobicity, tertiary structure changes and free sulfhydryl. The work provides a deep insight into the molecular conformation−emulsifiability relationship of EWP−monosaccharides conjugates.

Molecular Modification of Single Cobalt Sites Boosts the Catalytic Activity of CO2 Electroreduction into CO.

Electroreduction of CO2 into carbonaceous fuels or industrial chemicals using renewable energy sources is an ideal way to promote global carbon recycling. Thus, it is of great importance to develop highly selective, efficient, and stable catalysts. Herein, we prepared cobalt single atoms (Co SAs) coordinated with phthalocyanine (Co SAs-Pc). The anchoring of phthalocyanine with Co sites enabled electron transfer from Co sites to CO2 effectively via the π-conjugated system, resulting in high catalytic performance of CO2 electroreduction into CO. During the process of CO2 electroreduction, the Faradaic efficiency (FE) of Co SAs-Pc for CO was as high as 94.8 %. Meanwhile, the partial current density of Co SAs-Pc for CO was -11.3 mA cm-2 at -0.8 V versus the reversible hydrogen electrode (vs RHE), 18.83 and 2.86 times greater than those of Co SAs (-0.60 mA cm-2 ) and commercial Co phthalocyanine (-3.95 mA cm-2 ), respectively. In an H-cell system operating at -0.8 V vs RHE over 10 h, the current density and FE for CO of Co SAs-Pc dropped by 3.2 % and 2.5 %. A mechanistic study revealed that the promoted catalytic performance of Co SAs-Pc could be attributed to the accelerated reaction kinetics and facilitated CO2 activation.

Free-Radical-Mediated Formation of Trans-Cardiolipin Isomers, Analytical Approaches for Lipidomics and Consequences of the Structural Organization of Membranes.

Free-radical-mediated processes, such as peroxidation, isomerization and hydrogenation affecting fatty acid integrity and biological functions, have a trans-disciplinary relevance. Cardiolipins (CL, (1,3-diphosphatidyl-sn-glycerol)) and tetra-linoleoyl-CL are complex phospholipids, exclusively present in the Inner Mitochondrial Membrane (IMM) lipids, where they maintain membrane integrity and regulate enzyme functionalities. Peroxidation pathways and fatty acid remodeling are known causes of mitochondrial disfunctions and pathologies, including cancer. Free-radical-mediated isomerization with the change of the cis CL into geometrical trans isomers is an unknown process with possible consequences on the supramolecular membrane lipid organization. Here, the formation of mono-trans CL (MT-CL) and other trans CL isomers (T-CL) is reported using CL from bovine heart mitochondria and thiyl radicals generated by UV-photolysis from 2-mercaptoethanol. Analytical approaches for CL isomer separation and identification via 1H/13C NMR are provided, together with the chemical study of CL derivatization to fatty acid methyl esters (FAME), useful for lipidomics and metabolomics research. Kinetics information of the radical chain isomerization process was obtained using γ-irradiation conditions. The CL isomerization affected the structural organization of membranes, as tested by the reduction in unilamellar liposome diameter, and accompanied the well-known process of oxidative consumption induced by Fenton reagents. These results highlight a potential new molecular modification pathway of mitochondrial lipids with wide applications to membrane functions and biological consequences.

Quality assessment of DNA and hemoglobin by Fourier transform infrared spectroscopy in occupational exposure to extremely low-frequency magnetic field.

Previous studies have shown the effect of extremely low-frequency (ELF) magnetic fields on the hematopoietic system. However, molecular modification and biological toxicity are not known yet. The aim of this study was to investigate the effect of occupational exposure to ELF magnetic field on the hemoglobin and DNA alteration using Fourier transform infrared (FTIR) spectroscopy. Twenty nine individuals were selected among those working as the controller in a powerhouse in order to be studied as the population exposed to ELF magnetic field. Control group comprised of 29 administrative employees voluntarily participated who were matched with the exposed subjects in terms of sex, age, work experiences, smoking habit, and socioeconomic status. DNA and hemoglobin were extracted from blood samples and then were studied by FTIR spectroscopy. The results showed the level of magnetic field exposure was between 0.38 to 50 μT in the exposed subjects while the level of magnetic field exposure was between 0.19 and 20 μT for the unexposed people. Hemoglobin level was equal to 15.67 ± 1.42g/dL for exposed subjects which is significantly lower than that of the unexposed people (p = 0.0001). There was a significant alteration in CH content and COO structure of the hemoglobin structure. Moreover, DNA showed significant changes by functional group of organic base. This change in the structure of DNA and hemoglobin can lead to the creation of risks in human health. In conclusion, FTIR method could reveal the quality of DNA and hemoglobin structure in subjects after exposure to ELF magnetic field.

Effect of transient scrotal hyperthermia on human sperm: an iTRAQ-based proteomic analysis

BackgroundThrough this prospective study, we aimed to explore the change of molecular modification after the transient scrotal hyperthermia on human sperm.MethodsTen healthy subjects selected with strict screening criteria underwent testicular warming in a 43 °C water bath for 30 min a day for 10 consecutive days. Semen samples were collected 2 weeks before the first heat treatment and 6 weeks after the first heat treatment. Proteins from the samples were labeled with isobaric tags for relative and absolute quantitation and analyzed by two-dimensional liquid chromatography–tandem mass spectrometry.ResultsIn contrast to the control, of the 3446 proteins identified, 61 proteins were deregulated: 28 were up-regulated and 33 were down-regulated. Approximately 95% of the differentially expressed proteins were found to participate in spermatogenesis, fertilization, or other aspects of reproduction. In particular, the expression of sperm motility and energy metabolism-related proteins AKAP4, SPESP1, ODF1, ODF2, GAPDHS, and ACTRT2, validated by western blotting of the proteins obtained from human and mouse samples, tended to be reduced under scrotal hyperthermia.ConclusionsThe results indicated that the proteins AKAP4, ODF1, ODF2, GAPDHS, SPESP1, and ACTRT2, play an important role in the heat-induced reversible reduction in sperm concentration and motility and have the potential to be the biomarkers and clinical targets for scrotal heat treatment induced male infertility.

Drug discovery, explain how lead compound delivers to the target site and claim if there are recommended antiviral drugs to COVID-19.

Drug design is a long and costly process taking many stages; start with target identification passing through target validation, lead identification, and candidate optimization of pre-clinical and clinical trials. Drug design types depend on screening of a large number of molecules to distinguish and can select the most effective drug with high pharmaceutical effect. Ligand and Structure Based Drug Design are the two types of drug design. The drug takes journey when administered into the body through several ways (oral, inhalation, Intravenous (IV), Intramuscular (IM)) to deliver its target site. Twenty years ago, Computational strategies applied to understand particular target molecules with hits achieving lead target and that helps in lead Identification and Optimization stages of drug design and development. Screening, molecular modification and rational drug design are the three approaches to search for a modern drug. Bioinformatics plays a vital role in the discovery of drug as Bioinformatics includes both the programmed preparing of huge amounts of existing information and the creation of modern sorts of data resource. Both required in case the information is to be changed into data and utilized to assist in drug discovery. This work will discuss what is the meaning of drug design? What are the stages of drug design? What are the drug design types? Computer- Aided Drug Design (CAAD), Approaches in the search for a new drug, What is the journey of drug to deliver its target site? What are the methods of drug delivery? And will discuss the recommended antiviral therapy in the management of COVID-19.

A Multi-functional Molecular Modifier Enabling Efficient Large-Area Perovskite Light-Emitting Diodes

<h2>Summary</h2> With rapid progress in perovskite light-emitting diodes (PeLEDs), the electroluminescence performance of large-area is of increasing interest. We investigated why large-area performance lags behind that achieved in laboratory-scale devices and found that defects in perovskite films—emerging from thermal convection during solvent evaporation, as well as electronic traps formed during perovskite crystallization—are chief causes. Here, we report a molecular modification strategy that simultaneously eliminates pinholes in perovskite layers by controlling the dynamics of film formation and that passivates defects in perovskites by incorporating Br species, thereby preventing shorts and non-radiative recombination. The molecular modifier 1,3,5-tris (bromomethyl) benzene (TBB) also modulates the electronic structure of injection or transport materials to achieve improved charge injection and balanced charge transport. As a result, we demonstrate 20 mm × 20 mm green perovskite nanocrystal LEDs that achieve an external quantum efficiency (EQE) of over 16%, a record for large-area PeLEDs.

Construction of a Highly Diastereoselective Aldol Reaction System with l-Threonine Aldolase by Computer-Assisted Rational Molecular Modification and Medium Engineering.

Diastereoselectivity of l-threonine aldolase (LTA) was determined by paths of aldehydes attacking a pyridoxal phosphate-glycine complex. Thus, strategies of enhancing the syn path and blocking the anti path were performed to modify LTA. A mutant (Y31H/N305R) was constructed with a substrate preference increase from 3.32 to 42.04. Medium engineering was investigated. Consequently, the de value of l-syn-3-[4-(methylsulfonyl)phenylserine] reached 93.1% (87.2%conv). The study clarified the factors affecting diastereoselectivity of LTA and provided a theorem for rational modification of LTA's diastereoselectivity.

Expression optimization and molecular modification of heparin C5 epimerase

Heparin and heparan sulfate are a class of glycosaminoglycans for clinical anticoagulation. Heparosan N-sulfate-glucuronate 5-epimerase (C5, EC 5.1.3.17) is a critical modifying enzyme in the synthesis of heparin and heparan sulfate, and catalyzes the inversion of carboxyl group at position 5 on D-glucuronic acid (D-GlcA) of N-sulfoheparosan to form L-iduronic acid (L-IdoA). In this study, the heparin C5 epimerase gene Glce from zebrafish was expressed and molecularly modified in Escherichia coli. After comparing three expression vectors of pET-20b (+), pET-28a (+) and pCold Ⅲ, C5 activity reached the highest ((1 873.61±5.42) U/L) with the vector pCold Ⅲ. Then we fused the solution-promoting label SET2 at the N-terminal for increasing the soluble expression of C5. As a result, the soluble protein expression was increased by 50% compared with the control, and the enzyme activity reached (2 409±6.43) U/L. Based on this, site-directed mutations near the substrate binding pocket were performed through rational design, the optimal mutant (V153R) enzyme activity and specific enzyme activity were (5 804±5.63) U/L and (145.1±2.33) U/mg, respectively 2.41-fold and 2.28-fold of the original enzyme. Modification and expression optimization of heparin C5 epimerase has laid the foundation for heparin enzymatic catalytic biosynthesis.

Alkylated Sesamol Derivatives as Potent Antioxidants.

Sesamol is a phenolic derivative. Its antioxidant activity is low than that of Trolox and depends on benzodioxole moiety. Thus, a molecular modification strategy through alkylation, inspired by natural and synthetic antioxidants, was studied by molecular modeling at the DFT/B3LYP level of theory by comparing the 6-31+G(d,p) and 6-311++G(2d,2p) basis sets. All proposed derivatives were compared to classical related antioxidants such as Trolox, t-butylated hydroxytoluene (BHT) and t-butylated hydroxyanisole (BHA). According to our results, molecular orbitals, single electron or hydrogen-atom transfers, spin density distributions, and alkyl substitutions at the ortho positions related to phenol moiety were found to be more effective than any other positions. The trimethylated derivative was more potent than Trolox. t-Butylated derivatives were stronger than all other alkylated derivatives and may be new alternative forms of modified antioxidants from natural products with applications in the chemical, pharmaceutical, and food industries.

Unraveling genes promoting ROS metabolism in subcellular organelles of Oryza sativa in response to trivalent and hexavalent chromium

Plants possess a well-organized protective network, wherein antioxidant enzymes play an important part in dealing with oxidative stress induced by over accumulation of ROS in plant cells. In the present study, a microcosm hydroponic experiment was performed to investigate the molecular modification of antioxidant enzymes at subcellular levels in rice seedlings in the presence of either trivalent [Cr(III)] or hexavalent chromium [Cr(VI)] using rice oligonucleotide microarray analysis. The results indicated that the production of ROS induced by Cr(III, VI) was concentration-dependent, Cr-specific and tissue-specific. Trivalent or hexavalent chromium exposure significantly (p < 0.05) altered the antioxidant enzymes activities in both rice tissues in comparison to control plants. In total, 41 genes were identified from the data of rice oligonucleotide microarray analysis. Under Cr(III) exposure, relatively higher expression of genes was observed in roots compared to those in shoots (p < 0.05), while gene expressions in both plant parts differed slightly during Cr(VI) exposure, implying different regulation and response strategies of plants against Cr(III) and Cr(VI). Subcellular localization indicated that genes encoding SOD, POD, APX, and GPX are mainly prevalent in the cytoplasm (30.77%), chloroplasts (29.23%), peroxisomes (10.77%) and mitochondria (9.23%), suggesting that cytoplasm and chloroplasts are the main sites responsible for scavenging ROS through enzymatic processes. Our study provides new insight into the roles of antioxidant enzymes in ROS metabolism at subcellular levels under Cr exposure.

Expression, homology modeling and enzymatic characterization of a new \u03b2-mannanase belonging to glycoside hydrolase family 1 from Enterobacter aerogenes B19

Backgroundβ-mannanase can hydrolyze β-1,4 glycosidic bond of mannan by the manner of endoglycosidase to generate mannan-oligosaccharides. Currently, β-mannanase has been widely applied in food, medicine, textile, paper and petroleum exploitation industries. β-mannanase is widespread in various organisms, however, microorganisms are the main source of β-mannanases. Microbial β-mannanases display wider pH range, temperature range and better thermostability, acid and alkali resistance, and substrate specificity than those from animals and plants. Therefore microbial β-mannanases are highly valued by researchers. Recombinant bacteria constructed by gene engineering and modified by protein engineering have been widely applied to produce β-mannanase, which shows more advantages than traditional microbial fermentation in various aspects.ResultsA β-mannanase gene (Man1E), which encoded 731 amino acid residues, was cloned from Enterobacter aerogenes. Man1E was classified as Glycoside Hydrolase family 1. The bSiteFinder prediction showed that there were eight essential residues in the catalytic center of Man1E as Trp166, Trp168, Asn229, Glu230, Tyr281, Glu309, Trp341 and Lys374. The catalytic module and carbohydrate binding module (CBM) of Man1E were homologously modeled. Superposition analysis and molecular docking revealed the residues located in the catalytic module of Man1E and the CBM of Man1E. The recombinant enzyme was successfully expressed, purified, and detected about 82.5 kDa by SDS-PAGE. The optimal reaction condition was 55 °C and pH 6.5. The enzyme exhibited high stability below 60 °C, and in the range of pH 3.5–8.5. The β-mannanase activity was activated by low concentration of Co2+, Mn2+, Zn2+, Ba2+ and Ca2+. Man1E showed the highest affinity for Locust bean gum (LBG). The Km and Vmax values for LBG were 3.09 ± 0.16 mg/mL and 909.10 ± 3.85 μmol/(mL min), respectively.ConclusionsA new type of β-mannanase with high activity from E. aerogenes is heterologously expressed and characterized. The enzyme belongs to an unreported β-mannanase family (CH1 family). It displays good pH and temperature features and excellent catalysis capacity for LBG and KGM. This study lays the foundation for future application and molecular modification to improve its catalytic efficiency and substrate specificity.