Global-to-Local Dependencies in Phosphorus Mass Flows and Markets: Pathways to Improving System Resiliency in Response to Exogenous Shocks.

Uneven global distribution of phosphate rock deposits and the supply chains to transport phosphorus (P) make P fertilizers vulnerable to exogenous shocks, including commodity market shocks; extreme weather events or natural disasters; and geopolitical instability, such as trade disputes, disruption of shipping routes, and war. Understanding bidirectional risk transmission (global-to-local and local-to-global) in P supply and consumption chains is thus essential. Ignoring P system interdependencies and associated risks could have major impacts on critical infrastructure operations and increase the vulnerability of global food systems. We highlight recent unanticipated events and cascading effects that have impacted P markets globally. We discuss the need to account for exogenous shocks in local assessments of P flows, policies, and infrastructure design choices. We also provide examples of how accounting for undervalued global risks to the P industry can hasten the transition to a sustainable P future. For example, leveraging internal P recycling loops, improving plant P use efficiency, and utilizing legacy soil P all enhance system resiliency in the face of exogenous shocks and long-term anticipated threats. Strategies applied at the local level, which are embedded within national and global policy systems, can have global-scale impacts in derisking the P supply chain.

Enhanced perfluorooctanoic acid (PFOA) degradation by electrochemical activation of peroxydisulfate (PDS) during electrooxidation for water treatment.

Improved treatment of per- and polyfluoroalkyl substances (PFAS) in water is critically important in light of the proposed United States Environmental Protection Agency (USEPA) drinking water regulations at ng L-1 levels. The addition of peroxymonosulfate (PMS) during electrooxidation (EO) can remove and destroy PFAS, but ng L-1 levels have not been tested, and PMS itself can be toxic. The objective of this research was to test peroxydisulfate (PDS, an alternative to PMS) activation by boron-doped diamond (BDD) electrodes for perfluorooctanoic acid (PFOA) degradation. The influence of PDS concentration, temperature, and environmental water matrix effects, and PFOA concentration on PDS-EO performance were systematically examined. Batch reactor experiments revealed that 99 % of PFOA was degraded and 69 % defluorination was achieved, confirming PFOA mineralization. Scavenging experiments implied that sulfate radicals (SO4-) and hydroxyl radicals (HO) played a more important role for PFOA degradation than 1O2 or electrons (e-). Further identification of PFOA degradation and transformation products by liquid chromatography-mass spectrometry (LC-MS) analysis established plausible PFOA degradation pathways. The analysis corroborates that direct electron transfers at the electrode initiate PFOA oxidation and SO4- improves overall treatment by cleaving the CC bond between the C7F15 and COOH moieties in PFOA, leading to possible products such as C7F15 and F-. The perfluoroalkyl radicals can be oxidized by SO4- and HO, resulting in the formation of shorter chain perfluorocarboxylic acids (e.g., perfluorobutanoic acid [PFBA]), with eventual mineralization to CO2 and F-. At an environmentally relevant low initial concentration of 100 ng L-1 PFOA, 99 % degradation was achieved. The degradation of PFOA was slightly affected by the water matrix as less removal was observed in an environmental river water sample (91 %) compared to tests conducted in Milli-Q water (99 %). Overall, EO with PDS provided a destructive approach for the elimination of PFOA.

Mitigation of viruses of concern and bacteriophage surrogates via common unit processes for water reuse: A meta-analysis.

Water reuse is a growing global reality. In regulating water reuse, viruses have come to the fore as key pathogens due to high shedding rates, low infectious doses, and resilience to traditional wastewater treatments. To demonstrate the high log reductions required by emerging water reuse regulations, cost and practicality necessitate surrogates for viruses for use as challenge organisms in unit process evaluation and monitoring. Bacteriophage surrogates that are mitigated to the same or lesser extent than viruses of concern are routinely used for individual unit process testing. However, the behavior of these surrogates over a multi-barrier treatment train typical of water reuse has not been well-established. Toward this aim, we performed a meta-analysis of log reductions of common bacteriophage surrogates for five treatment processes typical of water reuse treatment trains: advanced oxidation processes, chlorination, membrane filtration, ozonation, and ultraviolet (UV) disinfection. Robust linear regression was applied to identify a range of doses consistent with a given log reduction of bacteriophages and viruses of concern for each treatment process. The results were used to determine relative conservatism of surrogates. We found that no one bacteriophage was a representative or conservative surrogate for viruses of concern across all multi-barrier treatments (encompassing multiple mechanisms of virus mitigation). Rather, a suite of bacteriophage surrogates provides both a representative range of inactivation and information about the effectiveness of individual processes within a treatment train. Based on the abundance of available data and diversity of virus treatability using these five key water reuse treatment processes, bacteriophages MS2, phiX174, and Qbeta were recommended as a core suite of surrogates for virus challenge testing.

Evaluating Sustainable Development Pathways for Protein- and Peptide-Based Bioadsorbents for Phosphorus Recovery from Wastewater.

Recovering phosphate (P) from point sources such as wastewater effluent is a priority in order to alleviate the impacts of eutrophication and implement a circular economy for an increasingly limited resource. Bioadsorbents featuring P-binding proteins and peptides offer exquisite P specificity and sensitivity for achieving ultralow P concentrations, i.e., <100 µg P L-1, a discharge limit that has been implemented in at least one treatment facility in nine U.S. states. To prioritize research objectives for P recovery in wastewater treatment, we compared the financial and environmental sustainability of protein/peptide bioadsorbents to those of LayneRT anion exchange resin. The baseline scenario (reflecting lab-demonstrated performance at a full-scale implementation) had costs that were 3 orders of magnitude higher than those for typical wastewater treatment. However, scenarios exploring bioadsorbent improvements, including increasing the P-binding capacity per unit volume by using smaller P-selective peptides and nanoparticle base materials and implementing reuse, dramatically decreased median impacts to $1.06 m-3 and 0.001 kg CO2 equiv m-3; these values are in line with current wastewater treatment impacts and lower than the median LayneRT impacts of $4.04 m-3 and 0.19 kg CO2 equiv m-3. While the financial viability of capturing low P concentrations is a challenge, incorporating the externalities of environmental impacts may provide a feasible path forward to motivate ultralow P capture.

Improved disinfection byproduct removal using a polysulfone membrane loaded with powdered activated carbon.

Powdered activated carbon was immobilized by casting it in a polysulfone polymer membrane, which was then tested for disinfection byproduct (chloroform) and bacteria (Escherichia coli) removal. The membrane prepared using 90% T20 carbon and 10% polysulfone (M20-90) provided a filtration capacity of 2783 L m-2 , adsorption capacity of 2.85 mg g-1 , and 95% chloroform removal in a 10 s empty bed contact time. Flaws and cracks on the membrane surface caused by the carbon particles appeared to reduce chloroform and E. coli removal. To overcome this challenge, up to six layers of the M20-90 membrane were overlapped, which improved chloroform filtration capacity by 94.6%, to 5416 L m-2 , and increased the adsorption capacity by 93.3%, to 5.51 mg g-1 . E. coli removal also increased from 2.5 logs reduction using a single membrane layer to 6.3 logs using six layers under 10 psi feed pressure. The filtration flux declined from 6.94 m3  m-2  day-1  psi-1 for a single layer (0.45 mm thick) to 1.26 m3  m-2  day-1  psi-1 for the six-layer membrane system (2.7 mm thick). This work demonstrated the feasibility of using powdered activated carbon immobilized on a membrane to improve chloroform adsorption and filtration capacity while simultaneously removing microbes. PRACTITIONER POINTS: Powdered activated carbon was immobilized on a membrane to improve chloroform adsorption and filtration capacity while simultaneously removing microbes. Membranes made with the smaller carbon particles (T20) delivered better chloroform adsorption performance. Use of multiple layers of the membrane further improved chloroform and Escherichia coli removal.

Effects of Regional Climate and BMP Type on Stormwater Nutrient Concentrations in BMPs: A Meta-Analysis.

Nutrient treatment performance of stormwater best management practices (BMPs) is highly variable. Improved nutrient management with BMPs requires a better understanding of factors that influence stormwater BMP treatment processes. We conducted a meta-analysis of vegetated BMPs in the International Stormwater BMP Database and compared influent and effluent nitrogen and phosphorus concentrations to quantify the BMP effect on nutrient management across climates. BMP effect on nutrient concentration change was compared between vegetated BMPs in wet and dry climates. We examined paired dissolved inorganic nitrogen (DIN), total nitrogen (TN), dissolved inorganic phosphorus (DIP), total phosphorus (TP), and combinations of these analytes as dissolved inorganic ratios and N:P ratios. Meta-analysis with subgroup analysis was used to determine differences between wet and dry climates and among vegetated BMP types. We found that across both wet and dry climates, BMPs leach DIP and TP, increase the fraction of dissolved inorganic P (DIP:TP), and decrease dissolved N:P ratios. Dry-climate BMPs leach DIP and TP more consistently and at a higher magnitude than wet-climate BMPs, and bioretention leaches more DIP than grass strips and swales. These findings generally align with biogeochemical cycling, differences in influent chemistry, and BMP design types and goals.

Adsorption of recalcitrant phosphorus compounds using the phosphate-selective binding-protein PstS.

Currently available wastewater phosphorus (P) treatment technologies target removal of reactive forms of P. Selective adsorption of more recalcitrant soluble non-reactive phosphorus (sNRP) can improve P removal and recovery. A phosphate-selective phosphate-binding protein (PBP), PstS, was immobilized onto NHS-activated beads to assess the ability of this novel bioadsorbent to remove (adsorb) and subsequently recover (desorb) a range of sNRP compounds. Four sNRP compounds representative of wastewater sNRP were selected for use in this study: phytic acid (PA), sodium triphosphate (TrP), beta-glycerol phosphate (BGP), and sodium hexametaphosphate (HMP). Using PBP, adsorption of all sNRP compounds was thermodynamically favorable. The PBP had nearly equivalent binding affinity for PA compared to PBP's typical target, orthophosphate, although it had less affinity for the other sNRP compounds. Adsorption followed pseudo-second order reaction kinetics, with 95% of maximum adsorption occurring within 4 min. This was substantially faster sNRP adsorption compared to other adsorbents in the literature. Adsorption was modeled using the Langmuir isotherm, reflecting that one phosphate molecule attached to one PBP binding site. Notably, this selective 1:1 attachment resulted in higher total P removal for sNRP molecules with high P content. The binding site lost activity with increasing pH, and as such, highest desorption was achieved at pH 12, making the system amenable to sNRP removal as well as controlled recovery.

Meta-analysis of the prevalence of dissolved organic nitrogen (DON) in water and wastewater and review of DON removal and recovery strategies.

Most wastewater removal and recovery processes primarily target dissolved inorganic nitrogen (DIN) species, leaving the untreated non-reactive dissolved organic nitrogen (DON) in the effluent. This DON fraction can account for a substantial part of the total nitrogen (N) load. We analyzed large datasets of N species and concentrations (with a focus on quantifying the fraction of DON) in surface water, ground water, and wastewater effluent across the United States. We then reviewed strategies to remove and recover DON based on results of a range of treatment technologies reported in the literature, including laboratory-scale up to full-scale operation in wastewater treatment plants. Our meta-analysis showed that DON concentrations are greatest in wastewater effluent followed by surface water and groundwater. The concentration of DON in wastewater effluent varied from 0.01 to 10.9 mg N/L (number of data points, n = 163), where the range in surface water was 0.002 to 14.3 mg N/L (n = 11,803). Organic N accounted for the majority of total N in 12.3% of wastewater effluent samples and 49.1% of surface waters. Our literature review showed that currently available wastewater treatment processes do not efficiently target DON removal nor recovery of the DON as a valuable product. One potential DON removal and recovery strategy is transforming DON into DIN, which is generally more easily removed and recovered. Transformation strategies reported in the literature include ozonation, UV/H2O2, and electrooxidation. However, as advanced oxidation processes are often energy- and cost-intensive, further research is needed to improve DON removal and recovery.

Fixed-bed column study of phosphate adsorption using immobilized phosphate-binding protein.

Bio-adsorption using high-affinity phosphate-binding proteins (PBP) has demonstrated effective phosphorus removal and recovery in batch-scale tests. Subsequent optimization of design and performance of fixed-bed column systems is essential for scaling up and implementation. Here, continuous-flow fixed-bed column tests were used to investigate the adsorption of inorganic phosphate (orthophosphate, Pi) using phosphate-binding proteins immobilized on resin (PBP-NHS) targeting Pi removal to ultra-low levels followed by recovery. Time to breakthrough decreased with higher influent Pi concentration, smaller bed volume, and higher influent flow rates. The Thomas and Yoon-Nelson breakthrough models adequately described PBP-NHS resin performance with a correlation coefficient of R2 > 0.95. The sharp S-shape of the breakthrough curves for both Pi-only solution and multi-ion solution indicated highly favorable and selective separation of Pi using PBP-NHS resin, beyond that achieved using LayneRT™, a commercial ion exchange resin. The Pi adsorption capacity of the PBP-NHS column was unaffected by competing anions, whereas capacity of the LayneRT™ column dropped by 20%. Tertiary wastewater effluent was also successfully treated in PBP-NHS column tests with a typical S-shaped breakthrough curve. Operating the fixed-bed column in multi-cycle mode evidenced the reusability of PBP-NHS resin with no significant decline in column performance. The results of this study contribute to efforts to scale up designs of PBP-NHS adsorption systems.

Making Waves: Biocatalysis and Biosorption: Opportunities and Challenges Associated with a New Protein-Based Toolbox for Water and Wastewater Treatment.

New water and wastewater treatment technologies are required to meet the demands created by emerging contaminants and resource recovery needs, yet technology development is a slow and uncertain process. Through evolution, nature has developed highly selective and fast-acting proteins that could help address these issues, but research and application have been limited, often due to assumptions about stability and economic feasibility. Here we highlight the potential advantages of cell-free, protein-based water and wastewater treatment processes (biocatalysis and biosorption), evaluate existing information about their economic feasibility, consider when a protein-based treatment process might be advantageous, and highlight key research needs.

Electro-oxidation to convert dissolved organic nitrogen and soluble non-reactive phosphorus to more readily removable and recoverable forms.

Conventional wastewater treatment processes cannot effectively remove dissolved organic nitrogen (DON) and soluble non-reactive phosphorus (sNRP), which can pose regulatory compliance challenges for total nitrogen and total phosphorus discharges. Moreover, DON and sNRP are not easily recoverable for beneficial reuse as part of the waste to resource paradigm. Conversion of DON and sNRP to more readily removable dissolved inorganic nitrogen (DIN) and soluble reactive phosphorus (sRP), respectively, will help meet stringent nutrient limits and facilitate nutrient recovery. In this study, electro-oxidation (EO) was evaluated for conversion of four DON compounds to DIN and five sNRP compounds to sRP. EO was more efficient and provided higher extents of conversion of the recalcitrant nutrient fractions compared to a more traditional advanced oxidation process, UV/H2O2. Direct electron transfer was likely the dominant oxidation mechanism for EO-based DON and sNRP conversion, with DON being more recalcitrant. Among the DON compounds tested, greater availability of primary amine (C-N bonds) yielded greater conversion compared to compounds with fewer primary amine or those with secondary amine (C-N-C bond). Among the sNRP compounds tested, those with P-O-C bonds (organic sNRP) converted more readily than those with P-O-P bonds (inorganic sNRP), presumably because cleavage of the latter bond requires greater energy. Using 30 min of EO treatment, the highest DON and sNRP compound conversion was 11.7 ± 0.09% for urea and 31.1 ± 0.75% for beta-glycerol phosphate. A similar extent of EO-based conversion of DON (6.41 ± 1.5%) and sNRP (32.7 ± 3.3%) was observed in real wastewater.

A Comparison of Porphyrin Photosensitizers in Photodynamic Inactivation of RNA and DNA Bacteriophages.

Effective broad-spectrum antiviral treatments are in dire need as disinfectants and therapeutic alternatives. One such method of disinfection is photodynamic inactivation, which involves the production of reactive oxygen species from dissolved oxygen in response to light-stimulated photosensitizers. This study evaluated the efficacy of functionalized porphyrin compounds for photodynamic inactivation of bacteriophages as human virus surrogates. A blue-light light emitting diode (LED) lamp was used to activate porphyrin compounds in aqueous solution (phosphate buffer). The DNA bacteriophages ΦX174 and P22 were more resistant to porphyrin TMPyP photodynamic inactivation than RNA bacteriophage fr, with increasing rates of inactivation in the order: ΦX174 << P22 << fr. Bacteriophage ΦX174 was therefore considered a resistant virus suitable for the evaluation of three additional porphyrins. These porphyrins were synthesized from TMPyP by inclusion of a central palladium ion (PdT4) and/or the addition of a hydrophobic C14 chain (PdC14 or C14). While the inactivation rate of bacteriophage ΦX174 via TMPyP was similar to previous reports of resistant viruses, ΦX174 inactivation increased by a factor of approximately 2.5 using the metalloporphyrins PdT4 and PdC14. The order of porphyrin effectiveness was TMPyP < C14 < PdT4 < PdC14, indicating that both Pd2+ ligation and C14 functionalization aided virus inactivation.

Editorial: Let's talk about P(ee).

PRACTITIONER POINTS: Phosphorus recycling and reuse are imperative, and the water industry has an important role to play in this effort. Technologies capable of removing phosphorus to ultra-low levels and subsequent recovery for phosphorus reuse are needed. Inorganic ion exchange resins and organic bioadsorbents are promising for phosphorus removal and recovery as part of the waste-to-resource paradigm.

Conversion of soluble recalcitrant phosphorus to recoverable orthophosphate form using UV/H2O2.

Soluble non-reactive phosphorus (sNRP), such as inorganic polyphosphates and organic P, is not effectively removed by conventional physicochemical processes. This can impede water resource reclamation facilities' ability to meet stringent total P regulations. This study investigated a UV/H2O2 advanced oxidation process (AOP) for converting sNRP to the more readily removable/recoverable soluble reactive P (sRP), or orthophosphate, form. Synthetic water spiked with four sNRP compounds (beta-glycerol phosphate, phytic acid, triphosphate, and hexa-meta phosphate) at varying H2O2 concentration, UV fluence, pH, and temperature was initially tested. These compounds represent simple, complex, organic, and inorganic forms of sNRP potentially found in wastewater. The efficiency of sNRP to sRP conversion depended on whether the sNRP compound was organic or inorganic and the complexity of its chemical structure. Using 1 mM H2O2 and 0.43 J/cm2 (pH 7.5, 22 °C), conversion of the simple organic beta-glycerol phosphate to sRP was 38.1 ± 2.9%, which significantly exceeded the conversion of the other sNRP compounds. Although conversion was achieved, the electrical energy per order (EEO) was very high at 5.2 × 103 ± 5.2 × 102 kWh/m3. Actual municipal wastewater secondary effluent, with sNRP accounting for 15% of total P, was also treated using UV/H2O2. No wastewater sNRP to sRP conversion was observed, ostensibly due to interference from wastewater constituents. Wastewater utilities that have difficulty meeting stringent P levels might be able to target simple organic sNRP compounds, though alternative processes beyond UV/H2O2 need to be explored to overcome interference from wastewater constituents and target more complex organic and inorganic sNRP compounds.

Electrocoagulation-electrooxidation for mitigating trace organic compounds in model drinking water sources.

In-situ water treatment can be accomplished using electrochemical treatments such as electrocoagulation (EC), which generates coagulants, and electrooxidation (EO), which generates oxidants (e.g., free chlorine and reactive oxygen species) via electrolysis using boron-doped diamond electrodes. In sequential EC-EO, EC can remove oxidant scavengers present in dissolved organic carbon (DOC), thereby improving the efficacy of downstream oxidation via EO. This study evaluated sequential EC-EO (and each process independently for comparison) for mitigating the trace organic compounds (TOrCs) acyclovir (ACY), trimethoprim (TMP), and benzyldimethyldecylammonium chloride (BAC-C10) in model groundwaters and surface waters. EO-only removed greater than 70% of ACY and TMP but negligible BAC-C10 in model groundwaters. In model surface waters, EO-only removed ∼55-75% BAC-C10, but had less removal for ACY and TMP (∼20-55%), primarily due to DOC interference. Sequential-EC-EO was investigated to better gauge the potential process improvement due to the addition of EC ahead of EO. EC removed 74 ± 7% DOC from model surface water and improved downstream EO treatment relative to EO-only by a factor of 3.4 for ACY, 1.7 for TMP, and 1.4 for BAC-C10. When treating model groundwater, EC-EO resulted in no improvement compared to EO-only for ACY and TMP. BAC-C10 removal was attributed to the particle separation step between EC and EO rather than electrochemical inputs. EO-only treatment was more energy efficient for model groundwater compared to model surface waters based on electrical energy per order (EEO) values. Sequential EC-EO further improved the energy efficiency for treating model river water.

Immobilized phosphate-binding protein can effectively discriminate against arsenate during phosphate adsorption and recovery.

There is a strong impetus to establish a circular phosphorus economy by securing internally renewable phosphate (Pi ) resources for use as agricultural fertilizers. Reversible Pi adsorption technologies such as ion exchange can remove and recover Pi from water/wastewater for reuse. However, existing reversible adsorbents cannot effectively discriminate against arsenate (As(V)) due to the similarity between As(V) and Pi chemical structure. If As(V) is co-recovered with Pi , the value of the recovered products for agricultural reuse is low. The objective of this study was to construct an immobilized phosphate-binding protein (PBP)-based Pi removal and recovery system and analyze its selectivity for Pi adsorption in the presence of As(V). A range of conditions was tested, including independent, sequential, and simultaneous exposure of the two oxyanions to immobilized PBP (PBP resin). The purity of the recovered Pi product was assessed after inducing controlled desorption of the adsorbed oxyanions at high pH (pH 12.5). Pi constituted more than 97% of the adsorbed oxyanions in the recovered product, even when As(V) was initially present at twofold higher concentrations than Pi . Therefore, PBP resin has potential to selectively remove Pi , as well as release high-purity Pi free of As(V) contamination suitable for subsequent agricultural reuse. PRACTITIONER POINTS: Existing reversible phosphate (Pi ) adsorbents cannot effectively discriminate against arsenate (As(V)) due to the similarity in their chemical structure. Co-recovery of As(V) with Pi can reduce the recovered product's reuse as a fertilizer. An immobilized phosphate-binding protein (PBP)-based system can be highly selective for Pi even in the presence of As(V). Pi constituted more than 97% of the recovered product, even when As(V) was present at 2-fold higher concentrations than Pi . Immobilized PBP offers advantages over existing Pi adsorbents by providing high-purity Pi products free of As(V) contamination for reuse.

Virus Isoelectric Point Estimation: Theories and Methods.

Much of virus fate, both in the environment and in physical/chemical treatment, is dependent on electrostatic interactions. Developing an accurate means of predicting virion isoelectric point (pI) would help to understand and anticipate virus fate and transport, especially for viruses that are not readily propagated in the lab. One simple approach to predicting pI estimates the pH at which the sum of charges from ionizable amino acids in capsid proteins approaches zero. However, predicted pIs based on capsid charges frequently deviate by several pH units from empirically measured pIs. Recently, the discrepancy between empirical and predicted pI was attributed to the electrostatic neutralization of predictable polynucleotide-binding regions (PBRs) of the capsid interior. In this paper, we review models presupposing (i) the influence of the viral polynucleotide on surface charge or (ii) the contribution of only exterior residues to surface charge. We then compare these models to the approach of excluding only PBRs and hypothesize a conceptual electrostatic model that aligns with this approach. The PBR exclusion method outperformed methods based on three-dimensional (3D) structure and accounted for major discrepancies in predicted pIs without adversely affecting pI prediction for a diverse range of viruses. In addition, the PBR exclusion method was determined to be the best available method for predicting virus pI, since (i) PBRs are predicted independently of the impact on pI, (ii) PBR prediction relies on proteome sequences rather than detailed structural models, and (iii) PBR exclusion was successfully demonstrated on a diverse set of viruses. These models apply to nonenveloped viruses only. A similar model for enveloped viruses is complicated by a lack of data on enveloped virus pI, as well as uncertainties regarding the influence of the phospholipid envelope on charge and ion gradients.

Improved Virus Isoelectric Point Estimation by Exclusion of Known and Predicted Genome-Binding Regions.

Knowledge of the isoelectric points (pIs) of viruses is beneficial for predicting virus behavior in environmental transport and physical/chemical treatment applications. However, the empirically measured pIs of many viruses have thus far defied simple explanation, let alone prediction, based on the ionizable amino acid composition of the virus capsid. Here, we suggest an approach for predicting the pI of nonenveloped viruses by excluding capsid regions that stabilize the virus polynucleotide via electrostatic interactions. This method was applied first to viruses with known polynucleotide-binding regions (PBRs) and/or three-dimensional (3D) structures. Then, PBRs were predicted in a group of 32 unique viral capsid proteome sequences via conserved structures and sequence motifs. Removing predicted PBRs resulted in a significantly better fit to empirical pI values. After modification, mean differences between theoretical and empirical pI values were reduced from 2.1 ± 2.4 to 0.1 ± 1.7 pH units.IMPORTANCE This model fits predicted pIs to empirical values for a diverse set of viruses. The results suggest that many previously reported discrepancies between theoretical and empirical virus pIs can be explained by coulombic neutralization of PBRs of the inner capsid. Given the diversity of virus capsid structures, this nonarbitrary, heuristic approach to predicting virus pI offers an effective alternative to a simplistic, one-size-fits-all charge model of the virion. The accurate, structure-based prediction of PBRs of the virus capsid employed here may also be of general interest to structural virologists.

Kinetics, Affinity, Thermodynamics, and Selectivity of Phosphate Removal Using Immobilized Phosphate-Binding Proteins.

A phosphate (Pi)-selective adsorption system featuring immobilized Pi-binding proteins (PBP) has recently attracted attention for ultralow Pi removal followed by recovery. This study investigated the adsorption kinetics, affinity, thermodynamics, and selectivity, as well as the effect of pH and temperature on Pi adsorption using immobilized PBP (PBP resin). Immobilizing PBP did not affect its Pi affinity. Kinetic studies at 22 °C and pH 7.1 showed that the PBP resin achieved 95% of its equilibrium capacity within 0.64 ± 0.2 min. The estimated Langmuir affinity constant (KL) was 21 ± 5 µM-1 Pi (220 ± 52 L/mg-Pi), which is higher than Pi adsorbents recently reported in literature. The ideal operating ranges for high-affinity Pi adsorption using PBP resin were pH 4.5 to 9 and temperature 14 to 37 °C. The Pi-PBP resin adsorption process was not affected by the presence of common anions (Cl-, Br-, NO2-, NO3-, SO42-, and HCO3-). Adsorption using the Pi-PBP resin was exothermic (ΔH = -6.3 ± 1.3 kJ/mol) and spontaneous (ΔG = -39.7 ± 0.1 to -43.2 ± 0.2 kJ/mol) between 14 and 43 °C. These results indicate that PBP resin's Pi adsorption rate and affinity surpass those of existing adsorbents. Future work to increase the PBP resin's adsorption capacity is important to its application as a viable Pi adsorbent.

Cell surface-expression of the phosphate-binding protein PstS: System development, characterization, and evaluation for phosphorus removal and recovery.

Simultaneous overabundance and scarcity of inorganic phosphate (Pi) is a critical issue driving the development of innovative water/wastewater treatment technologies that not only facilitate Pi removal to prevent eutrophication, but also recover Pi for agricultural reuse. Here, a cell-surface expressed high-affinity phosphate binding protein (PstS) system was developed, and its Pi capture and release potential was evaluated. E. coli was genetically modified to express PstS on its outer membrane using the ice nucleation protein (INP) as an anchoring motif. Verification of protein expression and localization were performed utilizing SDS-polyacrylamide gel electrophoresis (SDS-PAGE), western blot, and outer membrane separation analyses. Cell surface characterization was investigated through acid-base titration, X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). These tests provided information on the macromolecular structure and composition of the bacteria surface as well as the proton-exchange properties of the surface functional groups (i.e., pKa values). Phosphate desorption and adsorption batch experiments were conducted to evaluate the effects of temperature, pH, and ionic strength on phosphate capture and release. The PstS surface-displayed cells demonstrated greater potential to release and capture phosphate compared to non-modified cells. Higher temperatures up to 40°C, basic pH conditions (pH = 10.5), and higher ionic strength up to 1.0 mol/L KCl promoted 20%-50% higher phosphate release.