Molecular insight into arsenic uptake, transport, phytotoxicity, and defense responses in plants: a critical review.

MAIN CONCLUSION: A critical investigation into arsenic uptake and transportation, its phytotoxic effects, and defense strategies including complex signaling cascades and regulatory networks in plants. The metalloid arsenic (As) is a leading pollutant of soil and water. It easily finds its way into the food chain through plants, more precisely crops, a common diet source for humans resulting in serious health risks. Prolonged As exposure causes detrimental effects in plants and is diaphanously observed through numerous physiological, biochemical, and molecular attributes. Different inorganic and organic As species enter into the plant system via a variety of transporters e.g., phosphate transporters, aquaporins, etc. Therefore, plants tend to accumulate elevated levels of As which leads to severe phytotoxic damages including anomalies in biomolecules like protein, lipid, and DNA. To combat this, plants employ quite a few mitigation strategies such as efficient As efflux from the cell, iron plaque formation, regulation of As transporters, and intracellular chelation with an array of thiol-rich molecules such as phytochelatin, glutathione, and metallothionein followed by vacuolar compartmentalization of As through various vacuolar transporters. Moreover, the antioxidant machinery is also implicated to nullify the perilous outcomes of the metalloid. The stress ascribed by the metalloid also marks the commencement of multiple signaling cascades. This whole complicated system is indeed controlled by several transcription factors and microRNAs. This review aims to understand, in general, the plant-soil-arsenic interaction, effects of As in plants, As uptake mechanisms and its dynamics, and multifarious As detoxification mechanisms in plants. A major portion of this article is also devoted to understanding and deciphering the nexus between As stress-responsive mechanisms and its underlying complex interconnected regulatory networks.

The VirAB ABC Transporter Is Required for VirR Regulation of Listeria monocytogenes Virulence and Resistance to Nisin.

Listeria monocytogenes is a Gram-positive intracellular pathogen that causes a severe invasive disease. Upon infecting a host cell, L. monocytogenes upregulates the transcription of numerous factors necessary for productive infection. VirR is the response regulator component of a two-component regulatory system in L. monocytogenes In this report, we have identified the putative ABC transporter encoded by genes lmo1746-lmo1747 as necessary for VirR function. We have designated lmo1746-lmo1747 virAB We constructed an in-frame deletion of virAB and determined that the ΔvirAB mutant exhibited reduced transcription of VirR-regulated genes. The ΔvirAB mutant also showed defects in in vitro plaque formation and in vivo virulence that were similar to those of a ΔvirR deletion mutant. Since VirR is important for innate resistance to antimicrobial agents, we determined the MICs of nisin and bacitracin for ΔvirAB bacteria. We found that VirAB expression was necessary for nisin resistance but was dispensable for resistance to bacitracin. This result suggested a VirAB-independent mechanism of VirR regulation in response to bacitracin. Lastly, we found that the ΔvirR and ΔvirAB mutants had no deficiency in growth in broth culture, intracellular replication, or production of the ActA surface protein, which facilitates actin-based motility and cell-to-cell spread. However, the ΔvirR and ΔvirAB mutants produced shorter actin tails during intracellular infection, which suggested that these mutants have a reduced ability to move and spread via actin-based motility. These findings have demonstrated that L. monocytogenes VirAB functions in a pathway with VirR to regulate the expression of genes necessary for virulence and resistance to antimicrobial agents.

Linkage of the Nit1C gene cluster to bacterial cyanide assimilation as a nitrogen source.

A genetic linkage between a conserved gene cluster (Nit1C) and the ability of bacteria to utilize cyanide as the sole nitrogen source was demonstrated for nine different bacterial species. These included three strains whose cyanide nutritional ability has formerly been documented (Pseudomonas fluorescens Pf11764, Pseudomonas putida BCN3 and Klebsiella pneumoniae BCN33), and six not previously known to have this ability [Burkholderia (Paraburkholderia) xenovorans LB400, Paraburkholderia phymatum STM815, Paraburkholderia phytofirmans PsJN, Cupriavidus (Ralstonia) eutropha H16, Gluconoacetobacter diazotrophicus PA1 5 and Methylobacterium extorquens AM1]. For all bacteria, growth on or exposure to cyanide led to the induction of the canonical nitrilase (NitC) linked to the gene cluster, and in the case of Pf11764 in particular, transcript levels of cluster genes (nitBCDEFGH) were raised, and a nitC knock-out mutant failed to grow. Further studies demonstrated that the highly conserved nitB gene product was also significantly elevated. Collectively, these findings provide strong evidence for a genetic linkage between Nit1C and bacterial growth on cyanide, supporting use of the term cyanotrophy in describing what may represent a new nutritional paradigm in microbiology. A broader search of Nit1C genes in presently available genomes revealed its presence in 270 different bacteria, all contained within the domain Bacteria, including Gram-positive Firmicutes and Actinobacteria, and Gram-negative Proteobacteria and Cyanobacteria. Absence of the cluster in the Archaea is congruent with events that may have led to the inception of Nit1C occurring coincidentally with the first appearance of cyanogenic species on Earth, dating back 400-500 million years.

Bioaccumulation of cadmium by Enterobacter sp. and enhancement of rice seedling growth under cadmium stress.

Bacteria-mediated plant growth promotion and bioremediation of heavy metal containing soil is a widely accepted eco-friendly method. The present study is aimed to screen out cadmium resistant bacterial strain from metal contaminated rice rhizosphere and evaluate its effects on the growth of rice seedlings under cadmium stress. Among four different isolates (designated as S1, S2, S3 and S5), the S2 isolate was screened on the basis of different PGP traits and multi heavy metal resistance (minimum inhibitory concentration for cadmium, lead and arsenic were 3500, 2500 and 1050 µg/ml respectively). The selected S2 strain has ability to produce ACC deaminase (236.11 ng α-keto-butyrate/mg protein/h), IAA (726 µg/ml), solubilize phosphate (73.56 ppm) and fix nitrogen (4.4 µg of nitrogen fixed/h/mg protein). The selected strain was identified as Enterobacter sp. on the basis of phenotypic characterization, MALDI-TOF MS analysis of ribosomal proteins, FAME analysis and 16 S rDNA sequence homology. The high cadmium removal efficiency (> 95%) of this strain from the growth medium was measured by Atomic Absorption Spectrophotometer and it was due to intracellular cadmium accumulation evidenced by SEM-EDX-TEM-EDX study. SEM analysis also revealed no distortion of surface morphology of this strain even grown in the presence of high cadmium concentration (3000 µg/ml). Inoculation of this strain with rice seedlings significantly enhanced various morphological, biochemical characters of seedling growth compared with un-inoculated seedlings under Cd stress. The strain also exhibited alleviation of cadmium-induced oxidative stress, reduction of stress ethylene and decreased the accumulation of cadmium in seedlings as well that conferred cadmium tolerance to the plant. Thus the S2 strain could be considered as a potent heavy metal resistant PGPR applicable in heavy metal contaminated agricultural soil for bioremediation and plant growth promotion as well. MAIN FINDING: A cadmium resistant plant growth promoting Enterobacter sp. was isolated that accumulated cadmium evidenced by SEM-TEM-EDX study. It reduced Cd uptake and enhanced growth in rice seedlings.

Removal of diethyl phthalate from water by ozone microbubbles in a pilot plant.

Ozone microbubbles (OMBs) were used to remove diethyl phthalate (DEP) from water in a pilot plant. The removal of DEP and the mineralization efficiency were investigated under various reaction conditions. The removal of DEP by OMBs was very effective at the high pH and high ozone generation rates. Almost complete mineralization of DEP could be achieved at the high pH. The contribution of OH was computed by using a hydroxyl radical scavenger (i.e. t-BuOH). In neutral and alkaline media, the reaction of DEP with OH dominated over its direct reaction with ozone. The overall oxidation reaction fitted a second-order kinetic model. The overall rate constant and the volumetric mass transfer coefficient of ozone slightly increased with increasing pH. The results indicate that the OMBs were efficient in terms of the reduction of concentration of DEP and its complete mineralization.

Ecology and genomic features of infection with Mycobacterium avium subspecies paratuberculosis in Egypt.

Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is the causative agent of paratuberculosis, or Johne's disease, in cattle, with potential involvement in cases of Crohn's disease in humans. Johne's disease is found worldwide and is economically important for both beef and dairy industries. In an effort to characterize this important infection in Egypt, we analysed the ecological and genomic features of recent isolates of M. paratuberculosis. In this report, we examined 26 Holstein dairy herds distributed throughout Egypt, from 2010 to 2013. Using PCR analysis of faecal samples, we estimated a mean herd-level prevalence of 65.4 %, with animal-level infection that reached a mean of 13.6 % among animals suffering from diarrhoea. Whole genome sequencing of field isolates identified numerous single nucleotide polymorphisms among field isolates relative to the standard M. paratuberculosis K10 genome. Interestingly, the virulence of M. paratuberculosis isolates from Egypt revealed diverse virulence phenotypes in the murine model of paratuberculosis, with significant differences in tissue colonization, particularly during the chronic stage of infection. Overall, our analysis confirmed that Johne's disease is a newly identified problem in Egypt and indicated that M. paratuberculosis has potentially diverse genotypes that impact its virulence. Further ecological mapping and genomic analysis of M. paratuberculosis will enhance our understanding of the transmission and evolutionary dynamics of this pathogen under natural field conditions.

A pilot plant study of the degradation of Brilliant Green dye using ozone microbubbles: mechanism and kinetics of reaction.

Oxidation of Brilliant Green dye was performed using ozone microbubbles in a pilot plant scale. Decolourisation was very effective at both acidic and alkaline pH. The colour of the aqueous solution was below detectable limit after 30 min at 1.7 mg/s ozone generation rate. The reaction between the dye and ozone was first-order in nature with respect to both ozone and the dye. The enhancement factor increased with increasing dye concentration. The samples were analysed by the ultra-violet-visible (UV-Vis) spectrophotometry, gas chromatography-mass spectrometry (GC-MS) and Fourier transform infra-red (FTIR) spectroscopy. From the GC-MS analysis, 13 intermediates were detected as oxidation products of this dye at various stages of oxidation. The changes in the FTIR spectra showed the destruction of the dye and the formation of new compounds. The oxidation mechanism was divided into two reaction pathways. The mineralisation of Brilliant Green was up to 80% in 60 min, as determined by total organic carbon analysis.

Virulence and immunity orchestrated by the global gene regulator sigL in Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis causes Johne's disease in ruminants, a chronic enteric disease responsible for severe economic losses in the dairy industry. Global gene regulators, including sigma factors are important in regulating mycobacterial virulence. However, the biological significance of such regulators in M. avium subsp. paratuberculosis rremains elusive. To better decipher the role of sigma factors in M. avium subsp. paratuberculosis pathogenesis, we targeted a key sigma factor gene, sigL, activated in mycobacterium-infected macrophages. We interrogated an M. avium subsp. paratuberculosis ΔsigL mutant against a selected list of stressors that mimic the host microenvironments. Our data showed that sigL was important in maintaining bacterial survival under such stress conditions. Survival levels further reflected the inability of the ΔsigL mutant to persist inside the macrophage microenvironments. Additionally, mouse infection studies suggested a substantial role for sigL in M. avium subsp. paratuberculosis virulence, as indicated by the significant attenuation of the ΔsigL-deficient mutant compared to the parental strain. More importantly, when the sigL mutant was tested for its vaccine potential, protective immunity was generated in a vaccine/challenge model of murine paratuberculosis. Overall, our study highlights critical role of sigL in the pathogenesis and immunity of M. avium subsp. paratuberculosis infection, a potential role that could be shared by similar proteins in other intracellular pathogens.

Adjuvantation of a SARS-CoV-2 mRNA vaccine with controlled tissue-specific expression of an mRNA encoding IL-12p70.

Messenger RNA (mRNA) vaccines were pivotal in reducing severe acute respiratory syndrome 2 (SARS-CoV-2) infection burden, yet they have not demonstrated robust durability, especially in older adults. Here, we describe a molecular adjuvant comprising a lipid nanoparticle (LNP)-encapsulated mRNA encoding interleukin-12p70 (IL-12p70). The bioactive adjuvant was engineered with a multiorgan protection (MOP) sequence to restrict transcript expression to the intramuscular injection site. Admixing IL-12-MOP (CTX-1796) with the BNT162b2 SARS-CoV-2 vaccine increased spike protein-specific immune responses in mice. Specifically, the benefits of IL-12-MOP adjuvantation included amplified humoral and cellular immunity and increased immune durability for 1 year after vaccination in mice. An additional benefit included the restoration of immunity in aged mice to amounts comparable to those achieved in young adult animals, alongside amplification with a single immunization. Associated enhanced dendritic cell and germinal center responses were observed. Together, these data demonstrate that an LNP-encapsulated IL-12-MOP mRNA-encoded adjuvant can amplify immunogenicity independent of age, demonstrating translational potential to benefit vulnerable populations.

Key role for the alternative sigma factor, SigH, in the intracellular life of Mycobacterium avium subsp. paratuberculosis during macrophage stress.

Mycobacterium avium subsp. paratuberculosis causes Johne's disease, an enteric infection in cattle and other ruminants, greatly afflicting the dairy industry worldwide. Once inside the cell, M. avium subsp. paratuberculosis is known to survive harsh microenvironments, especially those inside activated macrophages. To improve our understanding of M. avium subsp. paratuberculosis pathogenesis, we examined phagosome maturation associated with transcriptional responses of M. avium subsp. paratuberculosis during macrophage infection. Monitoring cellular markers, only live M. avium subsp. paratuberculosis bacilli were able to prevent phagosome maturation and reduce its acidification. On the transcriptional level, over 300 M. avium subsp. paratuberculosis genes were significantly and differentially regulated in both naive and IFN-γ-activated macrophages. These genes include the sigma factor H (sigH) that was shown to be important for M. avium subsp. paratuberculosis survival inside gamma interferon (IFN-γ)-activated bovine macrophages. Interestingly, an sigH-knockout mutant showed increased sensitivity to a sustained level of thiol-specific oxidative stress. Large-scale RNA sequence analysis revealed that a large number of genes belong to the sigH regulon, especially following diamide stress. Genes involved in oxidative stress and virulence were among the induced genes in the sigH regulon with a putative consensus sequence for SigH binding that was recognized in a subset of these genes (n = 30), suggesting direct regulation by SigH. Finally, mice infections showed a significant attenuation of the ΔsigH mutant compared to its parental strain, suggesting a role for sigH in M. avium subsp. paratuberculosis virulence. Such analysis could identify potential targets for further testing as vaccine candidates against Johne's disease.

Reforming the countermeasures injury compensation program for COVID-19 and beyond: An economic perspective.

As of Aug. 2, 2021, 1693 injury claims associated with COVID-19 medical countermeasures have been filed in the Countermeasures Injury Compensation Program (CICP), of which 686 claims were related to COVID-19 vaccines and urgently needed compensation decisions. However, from an economic and public policy perspective, we find that the CICP design has unintended consequences: locating CICP in the executive agency DHHS potentially creates a conflict of interest, and not permitting judicial review generates a lack of checks and balances, both of which could jeopardize justice. These fundamental problems would subsequently weaken four key performance indicators of CICP compared with its judicial counterpart in the Court of Federal Claims. CICP lacks accountability, transparency, and cost-effectiveness efficiency, with 94% of its total costs spent on administration rather than compensation. CICP's ability to compensate is also questionable. If COVID-19 claims were compensated at its historical rate, CICP would face around $21.16 million in compensation outlays and $317.94 million in total outlays, 72.1 times its current balance. To ensure just compensation for injured petitioners during COVID-19 and future public health emergencies, we recommend Congress (1) initiate a major reform by relocating CICP from DHHS to the Claims Court or (2) keep CICP within DHHS and make incremental changes by permitting judicial review of DHHS administrative adjudication of CICP claims. We further recommend Congress audit and adjust budgets for CICP and DHHS promptly propose an injury table for COVID-19 claims. This is the first study that contributes an economic perspective to the limited literature on CICP and also provides unique and rich economic data.

Box-Cox power transformation unconditional quantile regressions with an application on wage inequality.

This study proposes a semi-parametric estimation method, Box-Cox power transformation unconditional quantile regression, to estimate the impact of changes in the distribution of the explanatory variables on the unconditional quantile of the outcome variable. The proposed method consists of running a nonlinear regression of the recentered influence function (RIF) of the outcome variable on the explanatory variables. We also show the asymptotic properties of the proposed estimator and apply the estimation method to address an existing puzzle in labor economics-why the 50th/10th percentile wage gap has been falling in the USA since the late 1980s. Our results show that declining unionization can explain approximately 10% of the decline in the 50/10 wage gap in 1990-2000 and 23% in 2000-2010.

Abatement of arsenic-induced phytotoxic effects in rice seedlings by an arsenic-resistant Pantoea dispersa strain.

Population detonation and rapid industrialization are the major factors behind the reduction in cultivable land that affects crop production seriously. This situation is further being deteriorated due to the negative effects of abiotic stresses. Under such conditions, plant growth-promoting rhizobacteria (PGPR) are found to improve crop production which is essential for sustainable agriculture. This study is focused on the isolation of potent arsenic (As)-resistant PGPR from the agricultural land of West Bengal, India, and its application to reduce As translocation in rice seedlings. After screening, an As-resistant PGPR strain AS18 was identified by phenotypic characters and 16S rDNA sequence-based homology as Pantoea dispersa. This strain displayed nitrogen fixation, phosphate solubilization, 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) activity, indole-3-acetic acid (IAA) production, in addition to As (III) resistance up to 3750 µg/mL. The As removal efficiency of this strain was up to 93.12% from the culture medium as evidenced by AAS. The bioaccumulation property of AS18 strain was further validated by TEM-EDAX-XRD-XRF-FTIR studies. This strain showed significant morpho-biochemical improvements including antioxidant enzymatic activities and As-minimization in plant (rice) cells. Thus, being an As-resistant potent PGPR, AS18 strain is expected to be applied in As-spiked agricultural fields for bioremediation and phytostimulation.

Treatment of a Pharmaceutical Industrial Effluent by a Hybrid Process of Advanced Oxidation and Adsorption.

In the present study, a combined approach of ozone-based advanced oxidation and adsorption by activated char was employed for the treatment of a pharmaceutical industrial effluent. Ozone is a selective oxidant, but the addition of H2O2 generated in situ hydroxyl radicals, which is a non-selective stronger oxidant than ozone. The effluent obtained from the pharmaceutical industry mainly contained anti-cancer drugs, anti-psychotic drugs, and some pain killers. The peroxone process had 75-88.5% chemical oxygen demand (COD) reduction efficiency at pH 5-11 in 3 h. Adsorption by activated char further reduced the COD to 85.4-92.7% for pH 5-11 in 2.5 h. All other water quality parameters were significantly decreased (>73% removal) during ozonation. The primary operational parameters (system pH and H2O2 concentration) were also varied, and their effects were analyzed. The pseudo-first-order rate constants for ozonation were calculated, and they were found to be in the range of 1.42 × 10-4 to 3.35 × 10-4 s-1 for pH 5-11. The kinetic parameters for adsorption were calculated for the pseudo-first-order, pseudo-second-order, and Elovich models. The fit of the pseudo-first-order kinetic model to the experimental data was the best.

Characterization of the pathogenesis and immune response to Listeria monocytogenes strains isolated from a sustained national outbreak.

Listeria monocytogenes is an intracellular pathogen responsible for listeriosis, a foodborne disease that can lead to life-threatening meningitis. The 2011 L. monocytogenes cantaloupe outbreak was among the deadliest foodborne outbreaks in the United States. We conducted in vitro and in vivo infection analyses to determine whether strains LS741 and LS743, two clinical isolates from the cantaloupe outbreak, differ significantly from the common laboratory strain 10403S. We showed that LS741 and LS743 exhibited increased virulence, characterized by higher colonization of the brain and other organs in mice. Assessment of cellular immune responses to known CD8+ T cell antigens was comparable between all strains. However, pre-existing immunity to 10403S did not confer protection in the brain against challenge with LS741. These studies provide insights into the pathogenesis of clinical isolates linked to the 2011 cantaloupe outbreak and also indicate that currently utilized laboratory strains are imperfect models for studying L. monocytogenes pathogenesis.

Listeria monocytogenes Infection of the Brain.

Listeria monocytogenes is an intracellular bacterial pathogen that is frequently associated with food-borne infection. The ability of L. monocytogenes to cross the blood-brain barrier (BBB) is concerning as it can lead to life-threatening meningitis and encephalitis. The BBB protects the brain microenvironment from various toxic metabolites and microbial pathogens found in the blood following infection, and therefore supports brain homeostasis. The mechanisms by which L. monocytogenes present in the bloodstream cross the BBB to cause brain infections are not fully understood and there is also a lack of a robust model system to study brain infections by L. monocytogenes. Here, we present a simple mouse infection model to determine whether bacteria have crossed the BBB and to quantitate the burden of bacteria that have colonized the brain in vivo. In this method, animals were infected intravenously with L. monocytogenes and were humanely euthanized by exposure to CO2 followed by cervical dislocation. Cardiac perfusion of the animals was performed prior to harvesting infected organs. Blood was collected before perfusion and the number of bacteria per organ or mL of blood was determined by plating dilutions of the blood or organ homogenates on agar plates and counting the number of colonies formed. This method can be used to study novel receptor-ligand interactions that enhance infection of the brain by L. monocytogenes and can be easily adapted for the study of multiple bacterial pathogens.

Alkali Metal Ion Partitioning with Calix[4]arene-benzo-crown-6 Ionophore in Acidic Medium: Insights from Experiments, Statistical Mechanical Framework, and Molecular Dynamics Simulations.

The current work reports the experimental and predicted interfacial behavior of metal ion extraction from aqueous phase-diluent system using a newly synthesized calix-benzo-crown-6 (CBCBGA) ionophore. Conductor-like screening model for real solvents was used to predict the selectivity at infinite dilution for the metal ion complexes in both aqueous and diluent phases. The selectivity for Cs+-CBCBGA extraction was found to be higher than that of other metal ions, namely, K+, Na+, and Rb+. This was confirmed by the experimental distribution coefficients obtained in the diluents system at 3 M HNO3 along with 0.01 M CBCBGA/organic solvents. The high selectivity of Cs+-CBCBGA complex over other complexes (K+, Rb+, and Na+) in nitrobenzene was also confirmed and validated by the highest occupied molecular orbital-lowest unoccupied molecular orbital energy gap (i.e., 0.13114 > 0.12411 > 0.11719 > 0.11561 eV) and interaction energy (i.e., -68.25 > -57.11 > -55.52 > -52.37 kcal/mol). The interaction and free energies of the extraction were found to increase with the dielectric constant of the organic solvents, namely, nitrobenzene > o-nitrophenyl hexyl ether > 1-octanol > chloroform. Overall, a higher selectivity of Cs+ ion over that of other metal ions (K+, Na+, and Rb+) was obtained for the newly synthesized CBCBGA ionophore in a radioactive waste solution.

Computational elucidation of phylogenetic, structural and functional characteristics of Pseudomonas Lipases.

Lipase (triacylglycerol acylhydrolase, EC 3.1.1.3) catalyzes tri-, di-, and monoacyl glycerol of fat into glycerol and fatty acids. It has important roles in the digestion of lipids in living organisms and industrially as laundry detergents along with proteases. The microbial lipases are more stable, active and economically feasible compared to plant and animal sources. Hence, much attention was given to the maximum production of the enzyme from the microbial sources. The phylogenetic analysis revealed that the amino acid sequence of lipase protein and their corresponding cDNA of Pseudomonas aeruginosa clustered with Pseudomonas stutzeri among different species of Pseudomonas, while P. aeruginosa PA1 clustered with P. aeruginosa SJTD-1 among different strains of P. aeruginosa. The lipase of P. aeruginosa PA1 was a monomeric, acidic and thermostable protein having a molecular weight ranging in between 32.72 to 34.89 kDa. The protein was abundant with random coils and alpha helices in its secondary structure. The tertiary model showed 96.310 score as an overall quality factor. Hence, this in silico study gives some useful information about the lipase protein without performing crystal structure assessment by X-ray Crystallography or NMR study in wet lab experiments which could be helpful for isolation and characterization of the enzyme in vitro.

Computational-based structural, functional and phylogenetic analysis of Enterobacter phytases.

Myo-inositol hexakisphosphate phosphohydrolases (i.e., phytases) are known to be a very important enzyme responsible for solubilization of insoluble phosphates. In the present study, Enterobacter phytases have characterized by different phylogenetic, structural and functional parameters using some standard bio-computational tools. Results showed that majority of the Enterobacter phytases are acidic in nature as most of the isoelectric points were under 7.0. The aliphatic indices predicted for the selected proteins were below 40 indicating their thermostable nature. The average molecular weight of the proteins was 48 kDa. The lower values of GRAVY of the said proteins implied that they have better interactions with water. Secondary structure prediction revealed that alpha-helical content was highest among the other forms such as sheets, coils, etc. Moreover, the predicted 3D structure of Enterobacter phytases divulged that the proteins consisted of four monomeric polypeptide chains i.e., it was a tetrameric protein. The predicted tertiary model of E. aerogenes (A0A0M3HCJ2) was deposited in Protein Model Database (Acc. No.: PM0080561) for further utilization after a thorough quality check from QMEAN and SAVES server. Functional analysis supported their classification as histidine acid phosphatases. Besides, multiple sequence alignment revealed that "DG-DP-LG" was the most highly conserved residues within the Enterobacter phytases. Thus, the present study will be useful in selecting suitable phytase-producing microbe exclusively for using in the animal food industry as a food additive.