Highly efficient recovery of phosphate and fluoride from phosphogypsum leachate: Selective precipitation and adsorption.

Phosphogypsum, a typical by-product in the phosphorus chemical industry, could generate a large amount of leachate containing phosphate and fluoride in the process of rainfall and long-term stacking, which not only causes serious environmental pollution, but also leads to a waste of resources. In this study, a united treatment of calcium hydroxide precipitation and lanthanum zeolite (La-ZFA) adsorption was proposed to achieve the recovery of phosphate and fluoride from phosphogypsum leachate. In phosphogypsum, most phosphorus could be leached except P in the residual occurrence form, while for fluoride, only water-soluble F could be effectively leached. The optimum leaching amounts of phosphate and fluoride were 22.59 and 4.64 mg/g, respectively, at liquid-solid ratio of 400:1, leaching time of 120 min, pH of 6.0, particle size of >200 mesh (<0.075 mm), and leaching temperature of 25°C. Using Ca(OH)2 as the precipitant, the phosphate could be precipitated selectively from phosphogypsum leachate by controlling pH and time, and the concentrations of it decreased significantly to 0.29 mg/L at pH 10.0, with a removal efficiency of 99.48%. XRD, SEM and Visual MINTEQ software analysis proved that the main component of the precipitate was hydroxyapatite (Ca5(PO4)3(OH)). After P precipitation, a series of sorbents for fluoride were investigated, and La-ZFA sorbent was chosen and utilized to recover the fluoride from the leachate through a cyclic fixed-bed column. The efficiency of La-ZFA was basically not affected by the high concentration sulfate, and it can selectively adsorb fluoride from phosphogypsum leachate, leading to a final fluoride concentration of 0.29 mg/L in the effluent. The characterization demonstrated that fluoride might be adsorbed onto the La-ZFA via ligand exchange with hydroxy groups. The proposed method in this study is expected to sequentially recover phosphate and fluorine from the leachate of phosphogypsum, and it has great guiding significance for resource utilization and management of phosphogypsum.

Modulating Elasticity of Liposome for Enhanced Cancer Immunotherapy.

Cancer immunotherapy has emerged as a promising approach to cancer treatment in recent years. The physical and chemical properties of nanocarriers are critical factors that regulate the immune activation of antigen-presenting cells (APCs) in the tumor microenvironment (TME). Herein, we extensively investigated the behavior of liposome nanoparticles (Lipo-NPs) with different elasticities, focusing on their interaction with immune cells and their transport mechanisms from tumors to tumor-draining lymph nodes (tdLNs). Successfully preparing Lipo-NPs with distinct elastic properties, their varied behaviors were observed, concerning immune cell interaction. Soft Lipo-NPs exhibited an affinity to cell membranes, while those with medium elasticity facilitated the cargo delivery to macrophages through membrane fusion. Conversely, hard Lipo-NPs enter macrophages via classical cellular uptake pathways. Additionally, it was noted that softer Lipo-NPs displayed superior transport to tdLNs in vivo, attributed to their deformable nature with lower elasticity. As a result, the medium elastic Lipo-NPs with agonists (cGAMP), by activating the STING pathway and enhancing transport to tdLNs, promoted abundant infiltration of tumor-infiltrating lymphocytes (TILs), leading to notable antitumor effects and extended survival in a melanoma mouse model. Furthermore, this study highlighted the potential synergistic effect of medium elasticity Lipo-NPs with immune checkpoint blockade (ICB) therapy in preventing tumor immune evasion. These findings hold promise for guiding immune-targeted delivery systems in cancer immunotherapy, particularly in vaccine design for tdLNs targeting and eradicating metastasis within tdLNs.

Enhancing biofilm formation with powder carriers for efficient nitrogen and phosphorus removal.

This study assesses the improvement in nitrogen and phosphorus removal from wastewater achieved through the integration of zeolite and attapulgite carrier materials into the activated sludge (AS) process. It was found that the addition of these materials significantly enhanced the processing performance of the reactor. Specifically, the use of zeolite and attapulgite powders increased sludge particle sizes to averages of 231.56 µm and 219.62 µm, respectively. This facilitated micro-granule formation, substantially improving the settling characteristics of the sludge and boosting the activity and proliferation of essential microbes. Illumina MiSeq sequencing demonstrated significant accumulations of DGAOs (Candidatus_Competibacter) and DPAOs (Candidatus_Accumulibacter). Furthermore, these carriers augmented the protein content in extracellular polymers, enhancing the hydrophobicity of the sludge and promoting aggregation. Comparative analysis based on the extended Derjaguin, Landau, Verwey, and Overbeek (DLVO) theory indicated a preferential adhesion affinity of sludge for zeolite compared to attapulgite, attributed primarily to Lewis acid-base and electric double-layer interactions. These findings underscore zeolite's enhanced efficacy in biomass fixation and suggest significant potential for the technological advancement of wastewater treatment plants.

Can internet use promote farmers to adopt chemical fertilizer reduction and efficiency enhancement technology in China?-an empirical analysis based on endogenous switching probit model.

This study examines the impact of internet usage on farmer's adoption behavior of fertilizer reduction and efficiency enhancement technologies in China. Based on 1,295 questionnaires in Henan Province, this study constructs a counterfactual analysis framework and used endogenous switching probit model to analyze the effects and pathways of internet usage on farmer's adoption behavior of chemical fertilizer reduction and efficiency enhancement technologies. The results indicate that. (1) The proportion of farmers adopting chemical fertilizer reduction and efficiency enhancement technologies is 60.15%, while the proportion of farmers not adopting these technologies is 39.85%. (2) Internet usage directly influences farmers' adoption of fertilizer reduction and efficiency enhancement technologies. According to counterfactual assumption analysis, if farmers who currently use the Internet were to stop using it, the probability of them adopting these technologies would decrease by 28.09%. Conversely, for farmers who do not currently use the Internet, if they were to start using it, the probability of them adopting fertilizer reduction and efficiency enhancement technologies would increase by 40.67%. (3) Internet usage indirectly influences farmers' adoption behavior through mediating pathways of expected benefits and risk perception. In addition, social networks negatively moderate the impact of internet usage on farmers' behavior of chemical fertilizer reduction and efficiency enhancement technologies.

Convergent and divergent transcriptional reprogramming of motor and sensory neurons underlying response to peripheral nerve injury.

INTRODUCTION: Motor neurons differ from sensory neurons in aspects including origins and surrounding environment. Understanding the similarities and differences in molecular response to peripheral nerve injury (PNI) and regeneration between sensory and motor neurons is crucial for developing effective drug targets for CNS regeneration. However, genome-wide comparisons of molecular changes between sensory and motor neurons following PNI remains limited. OBJECTIVES: This study aims to investigate genome-wide convergence and divergence of injury response between sensory and motor neurons to identify novel drug targets for neural repair. METHODS: We analyzed two large-scale RNA-seq datasets of in situ captured sensory neurons (SNs) and motoneurons (MNs) upon PNI, retinal ganglion cells and spinal cord upon CNS injury. Additionally, we integrated these with other related single-cell level datasets. Bootstrap DESeq2 and WGCNA were used to detect and explore co-expression modules of differentially expressed genes (DEGs). RESULTS: We found that SNs and MNs exhibited similar injury states, but with a delayed response in MNs. We identified a conserved regeneration-associated module (cRAM) with 274 shared DEGs. Of which, 47% of DEGs could be changed in injured neurons supported by single-cell resolution datasets. We also identified some less-studied candidates in cRAM, including genes associated with transcription, ubiquitination (Rnf122), and neuron-immune cells cross-talk. Further in vitro experiments confirmed a novel role of Rnf122 in axon growth. Analysis of the top 10% of DEGs with a large divergence suggested that both extrinsic (e.g., immune microenvironment) and intrinsic factors (e.g., development) contributed to expression divergence between SNs and MNs following injury. CONCLUSIONS: This comprehensive analysis revealed convergent and divergent injury response genes in SNs and MNs, providing new insights into transcriptional reprogramming of sensory and motor neurons responding to axonal injury and subsequent regeneration. It also identified some novel regeneration-associated candidates that may facilitate the development of strategies for axon regeneration.

Biogenic Palladium Improved Perchlorate Reduction during Nitrate Co-Reduction by Diverting Electron Flow in a Hydrogenotrophic Biofilm.

Microbial reduction of perchlorate (ClO4-) is emerging as a cost-effective strategy for groundwater remediation. However, the effectiveness of perchlorate reduction can be suppressed by the common co-contamination of nitrate (NO3-). We propose a means to overcome the limitation of ClO4- reduction: depositing palladium nanoparticles (Pd0NPs) within the matrix of a hydrogenotrophic biofilm. Two H2-based membrane biofilm reactors (MBfRs) were operated in parallel in long-term continuous and batch modes: one system had only a biofilm (bio-MBfR), while the other incorporated biogenic Pd0NPs in the biofilm matrix (bioPd-MBfR). For long-term co-reduction, bioPd-MBfR had a distinct advantage of oxyanion reduction fluxes, and it particularly alleviated the competitive advantage of NO3- reduction over ClO4- reduction. Batch tests also demonstrated that bioPd-MBfR gave more rapid reduction rates for ClO4- and ClO3- compared to those of bio-MBfR. Both biofilm communities were dominated by bacteria known to be perchlorate and nitrate reducers. Functional-gene abundances reflecting the intracellular electron flow from H2 to NADH to the reductases were supplanted by extracellular electron flow with the addition of Pd0NPs.

Atomically dispersed copper-zinc dual sites anchored on nitrogen-doped porous carbon toward peroxymonosulfate activation for degradation of various organic contaminants.

Single-atom catalysts (SACs) have been widely studied in Fenton-like reactions, wherein their catalytic performance could be further enhanced by adjusting electronic structure and regulating coordination environment, although relevant research is rarely reported. This text elucidates fabrication of dual atom catalyst systems aimed at augmenting their catalytic efficiency. Herein, atomically dispersed copper-zinc (Cu-Zn) dual sites anchored on nitrogen (N)-doped porous carbon (NC), referred to as CuZn-NC, were synthesized using cage-encapsulated pyrolysis and host-guest strategies. The CuZn-NC catalyst exhibited high activity in activation of peroxymonosulfate (PMS) for degradation of organic pollutants. Based on synergistic effects of adjacent Cu and Zn atom pairs, CuZn-NC (PMS) system achieved 94.44 % bisphenol A (BPA) degradation in 24 min. The radical pathway predominated, and coexistence of non-radical species was demonstrated for BPA degradation in CuZn-NC/PMS system. More importantly, CuZn-NC/PMS system showed generality for degradation of various refractory contaminants. Our experiments indicate that CuZn-N sites on CuZn-NC act as active sites for bonding PMS molecules with optimal binding energy, while pyrrolic N sites are considered as adsorption sites for organic molecules. Overall, this research designs diatomic site catalysts (DACs), with promising implications for wastewater treatment.

PD-1 inhibitor combined with paclitaxel and cisplatin in the treatment of recurrent and metastatic hypopharyngeal/laryngeal squamous cell carcinoma: efficacy and survival outcomes.

Objective: This retrospective study analyzed the efficacy of PD-1 inhibitors combined with albumin-bound paclitaxel and cisplatin (TP regimen) in the treatment of recurrent and metastatic hypopharyngeal/laryngeal squamous cell carcinoma (RMHSCC/RMLSCC). Methods: Patients diagnosed and treated at the Sun Yat-sen University Cancer Center from August 1, 2020, to August 15, 2023, with histologically confirmed RMHSCC/RMLSCC were included. All patients received PD-1 inhibitors combined with albumin-bound paclitaxel (260mg/m2) and cisplatin (60mg/m2) for 3-4 cycles. The primary endpoints were overall survival (OS) and progression-free survival (PFS). Results: A total of 50 patients with RMHSCC/RMLSCC who received TP+PD-1 inhibitor therapy were included, with an objective response rate (ORR) of 56.0% (28/50). The 1-year and 2-year OS rates were 80.2% (95% CI: 69.3%-92.9%) and 68.6% (95% CI: 52.6%-89.5%), respectively, while the 1-year and 2-year PFS rates were 44.7% (95% CI: 31.9%-62.5%) and 26.0% (95% CI: 12.6%-53.4%), respectively. Treatment-related adverse events mainly included rash, myelosuppression, gastrointestinal reactions, and hypothyroidism. Conclusion: In the treatment of RMHSCC/RMLSCC with TP + PD-1 inhibitors, survival rates of patients can be improved while ensuring the safety of the treatment regimen.

TERC promotes non-small cell lung cancer progression by facilitating the nuclear localization of TERT.

The core of telomerase consists of the protein subunit telomerase reverse transcriptase (TERT) and the telomerase RNA component (TERC). So far, the role of TERC in cancer development has remained elusive. Here, we found TERC expression elevated in non-small cell lung cancer (NSCLC) tissues, which was associated with disease progression and poor prognosis in patients. Using NSCLC cell lines and xenograft models, we showed that knockdown of TERC caused cell cycle arrest, and inhibition of cell proliferation and migration. Mechanistically, TERC was exported to the cytoplasm by nuclear RNA export factor 1 (NXF1), where it mediated the interaction of TERT with other telomerase subunits. Depletion of TERC hindered the assembly and subsequent nuclear localization of the telomerase complex, preventing TERT from functioning in telomere maintenance and transcription regulation. Our findings suggest that TERC is a potential biomarker for NSCLC diagnosis and prognosis and can be a target for NSCLC treatment.

Earth-abundant Zn-dipyrrin chromophores for efficient CO2 photoreduction.

The development of strong sensitizing and Earth-abundant antenna molecules is highly desirable for CO2 reduction through artificial photosynthesis. Herein, a library of Zn-dipyrrin complexes (Z-1-Z-6) are rationally designed via precisely controlling their molecular configuration to optimize strong sensitizing Earth-abundant photosensitizers. Upon visible-light excitation, their special geometry enables intramolecular charge transfer to induce a charge-transfer state, which was first demonstrated to accept electrons from electron donors. The resulting long-lived reduced photosensitizer was confirmed to trigger consecutive intermolecular electron transfers for boosting CO2-to-CO conversion. Remarkably, the Earth-abundant catalytic system with Z-6 and Fe-catalyst exhibits outstanding performance with a turnover number of >20 000 and 29.7% quantum yield, representing excellent catalytic performance among the molecular catalytic systems and highly superior to that of noble-metal photosensitizer Ir(ppy)2(bpy)+ under similar conditions. Experimental and theoretical investigations comprehensively unveil the structure-activity relationship, opening up a new horizon for the development of Earth-abundant strong sensitizing chromophores for boosting artificial photosynthesis.

Phosphorus immobilization/release behavior of lanthanum-modified bentonite amended sediment under the dual effects of pH and dissolved organic carbon.

Lanthanum modified bentonite (LMB) is typical P-inactivating agent that has been applied in over 200 lakes. Dissolved organic carbon (DOC) and high pH restrict the phosphorus (P) immobilization performance of LMB. However, the P immobilization/release behaviors of LMB-amended sediment when suspended to overlying water with high pH and DOC have not yet been studied. In the present work, batch adsorption and long-term incubation experiments were performed to study the combined effects of pH and DOC on the P control by LMB. The results showed that the coexistence of low concentration of DOC or preloading with some DOC had a negligible effect on P binding by LMB. In the presence of DOC, the P adsorption was more pronounced at pH 7.5 and was measurably less at pH 9.5. Additionally, the pH value was the key factor that decided the P removal at low DOC concentration. The increase in pH and DOC could significantly promote the release of sediment P with a higher EPC0. Under such condition, a higher LMB dosage was needed to effectively control the P releasing from sediment. In sediment/water system with intermittent resuspension, the alkaline conditions greatly facilitated the release of sediment P and DOC, which increased from 0.087 to 0.581 mg/L, and from 11.05 to 26.56 mg/L, respectively. Under the dual effect of pH and DOC, the P-immobilization performance of LMB was weakened, and a tailor-made scheme became essential for determining the optimum dosage. The desorption experiments verified that the previously loaded phosphorus on LMB was hard to be released even under high pH and DOC conditions, with an accumulative desorption rate of less than 2%. Accordingly, to achieve the best P controlling efficiency, the application strategies depending on LMB should avoid the high DOC loading period such as the rainy season and algal blooms.

FLI1 promotes IFN-γ-induced kynurenine production to impair anti-tumor immunity.

Nasopharyngeal carcinoma (NPC)-mediated immunosuppression within the tumor microenvironment (TME) frequently culminates in the failure of otherwise promising immunotherapies. In this study, we identify tumor-intrinsic FLI1 as a critical mediator in impairing T cell anti-tumor immunity. A mechanistic inquiry reveals that FLI1 orchestrates the expression of CBP and STAT1, facilitating chromatin accessibility and transcriptional activation of IDO1 in response to T cell-released IFN-γ. This regulatory cascade ultimately leads to augmented IDO1 expression, resulting in heightened synthesis of kynurenine (Kyn) in tumor cells. This, in turn, fosters CD8+ T cell exhaustion and regulatory T cell (Treg) differentiation. Intriguingly, we find that pharmacological inhibition of FLI1 effectively obstructs the CBP/STAT1-IDO1-Kyn axis, thereby invigorating both spontaneous and checkpoint therapy-induced immune responses, culminating in enhanced tumor eradication. In conclusion, our findings delineate FLI1-mediated Kyn metabolism as an immune evasion mechanism in NPC, furnishing valuable insights into potential therapeutic interventions.

A practical study on the near-zero discharge of rainwater and the collaborative treatment and regeneration of rainwater and sewage.

Non-conventional water recovery, recycling, and reuse have been considered imperative approaches to addressing water scarcity in China. The objective of this study was to evaluate the technical and economic feasibility of Water Reclamation Plants (WRP) based on an anaerobic-anoxic-oxic membrane bioreactor (A2O-MBR) system for unconventional water resource treatment and reuse in towns (domestic sewage and rainwater). Rainwater is collected and stored in the rainwater reservoir through the rainwater pipe network, and then transported to the WRP for treatment and reuse through the rainwater reuse pumping station during the peak water demand period. During a year of operation and evaluation process, a total of 610,000 cubic meters of rainwater were reused, accounting for 10.4 % of the treated wastewater. In the A2O-MBR operation, the average effluent concentrations for COD (chemical oxygen demand), NH4+-N (ammonium), TN (total nitrogen), and TP (total phosphorus) were 14.23 ± 4.07 mg/L, 0.22 ± 0.26 mg/L, 11.97 ± 1.54 mg/L, and 0.13 ± 0.09 mg/L, respectively. The effluent quality met standards suitable for reuse in industrial cooling water or for direct discharge. The WRP demonstrates a positive financial outlook, with total capital and operating costs totaling 0.16 $/m3. A comprehensive cost-benefit analysis indicates a positive net present value for the WRP, and the estimated annualized net profit is 0.024 $/m3. This research has achieved near-zero discharge of wastewater and effective allocation of rainwater resources across time and space.

Fermentation broth from fruit and vegetable waste works: Reducing the risk of human bacterial pathogens in soil by inhibiting quorum sensing.

Fermentation broth from fruit and vegetable waste (FFVW) has demonstrated remarkable ability as a soil amendment and in reducing antibiotic resistance genes (ARGs) pollution. However, the potential of FFVW to mitigate other microbial contamination such as human bacterial pathogens (HBPs) and virulence factor genes (VFGs), which are closely associated with human health, remains unknown. In this study, metagenomic analysis revealed that FFVW reduced the HBPs with high-risk of ARGs and VFGs including Klebsiella pneumoniae (reduced by 40.4 %), Mycobacterium tuberculosis (reduced by 21.4 %) and Streptococcus pneumoniae (reduced by 38.7 %). Correspondingly, VFG abundance in soil decreased from 3.40 copies/cell to 2.99 copies/cell. Further analysis illustrated that these was mainly attributed to the inhibition of quorum sensing (QS). FFVW reduced the abundance of QS signals, QS synthesis genes such as rpaI and luxS, as well as receptor genes such as rpfC and fusK, resulting in a decreased in risk of ARGs and VFGs. The pure culture experiment revealed that the expression of genes related to QS, VFGs, ARGs and mobile genetic elements (MGEs) were downregulated in Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli and K. pneumoniae treated by FFVW, consistent with the result of metagenomic analysis. This study suggested an environmentally friendly approach for controlling soil VFGs/ARGs-carrying HBPs, which is crucial for both soil and human health under the framework of "One Health".

Intranasal Administration of Umbilical Cord Mesenchymal Stem Cell Exosomes Alleviates Parkinson's Disease.

Parkinson's disease (PD) is a common and complex neurodegenerative disease. This disease is typically characterized by the formation of Lewy bodies in multiple brain regions and dopaminergic neuronal loss in the substantia nigra pars compacta, resulting in non-motor symptoms (e.g., olfactory deficits) and motor dysfunction in the late stages. There is yet no effective cure for Parkinson's disease. Considering the neuroprotective effects of exosomes, we investigated whether intranasal administration of umbilical cord mesenchymal stem cell exosomes could improve behavioral functions in PD mice. First, exosomes were endocytosed by the cells in vitro and in vivo, indicating that exosomes can cross the blood-brain barrier. Second, we found that both motor and non-motor functions of the PD models were effectively improved during intranasal exosomes treatment. Finally, the activity of olfactory bulb neurons was improved and the loss of dopaminergic neurons in the substantia nigra pars compacta was reversed. Moreover, exosomes attenuated microglia and astrocyte activation, leading to a low level of inflammation in the brain. In conclusion, our study provided a new reference for the clinical application of exosomes in the treatment of PD.

Potential of free nitrous acid (FNA) for sludge treatment and resource recovery from waste activated sludge: A review.

The escalating production of waste activated sludge (WAS) presents significant challenges to wastewater treatment plants (WWTPs). Free nitrous acid (FNA), known for its biocidal effect, has gained a growing focus on sludge dewatering, sludge reduction, and resource recovery from WAS due to its eco-friendly and cost-effective properties. Nevertheless, there have been no attempts made to systematically summarize or critically analyze the application of FNA in enhancing treatment and resource utilization of sludge. In this paper, we provided an overview of the current understanding regarding the application potential and influencing factors of FNA in sludge treatment, with a specific focus on enhancing sludge dewatering efficiency and reducing volume. To foster resource development from sludge, various techniques based on FNA have recently been proposed, which were comprehensively reviewed with the corresponding mechanisms meticulously discussed. The results showed that the chemical oxidation and interaction with microorganisms of FNA played the core role in improving resource utilization. Furthermore, current challenges and future prospects of the FNA-based applications were outlined. It is expected that this review can refine the theoretical framework of FNA-based processes, providing a theoretical foundation and technical guidance for the large-scale demonstration of FNA.

Input-to-State Stability of Switched Network Control Systems Under Unknown Deception Attacks.

This article focuses on the stability issue of switched network control systems (SNCSs) under deception attacks described by a Bernoulli process with unknown probability distribution. The false information in deception attacks is unknown but bounded and may be state dependent or state independent. By means of the input-to-state stability (ISS) tool and the convex combination method, an improved lemma is first developed for SNCSs, which facilitates the derivations of our results. After that, some attack-independent sufficient conditions for the ISS of SNCSs are obtained for mode-dependent average dwell time switching and stochastic switching, respectively. Different from existing results, the concerned switching contributes to the stability of SNCSs, which benefits the ISS performance of SNCSs even though the unknown deception attacks cause all subsystems to be non-ISS. The proposed results provide an effective solution with strong robustness to deal with unknown deception attacks or denial-of-service attacks.

Feasibility of in-situ sludge fermentation coupled with partial denitrification: Key roles of initial organic matters and alkaline pH.

Achieving partial denitrification (PD) by using fermentation products extracted from waste activated sludge (WAS) rather than commercial organic matters is a promising approach for providing nitrite for anammox, while sludge reduction could also be realized by WAS reutilization. This study proposed an In-situ Sludge Fermentation coupled with Partial Denitrification (ISFPD) system and explored its performance under different conditions, including initial pH, nitrate concentrations, and organic matters. Results showed that nitrite production increased with the elevation of initial pH (from 6 to 9), and the highest nitrate-to-nitrite transformation ratio (NTR) reached 77% at initial pH 9. The PD rates and NTR were observed to be minimally influenced by initial nitrate concentrations. Acetate was preferred by denitrifying bacteria, while macromolecules such as proteins necessitated be hydrolyzed to be suitable for further utilization. The insights gained through this study paved the way for efficient nitrite production and sustainable WAS reutilization in harmony.

Development of a Universal One-Step Purification and Activation Method to Engineer Protein-Glutaminase through Rational Design.

Cytotoxic enzymes often exist as zymogens containing prodomains to keep them in an inactive state. Protein-glutaminase (PG), which can enhance various functional characteristics of food proteins, is an enzyme containing pro-PG and mature-PG (mPG). However, poor activity and stability limit its application while tedious purification and activation steps limit its high-throughput engineering. Here, based on structural analysis, we replaced the linker sequence between pro-PG and mPG with the HRV3C protease recognition sequence and then coexpressed it with HRV3C protease in Escherichia coli to develop an efficient one-step purification and activation method for PG. We then used this method to obtain several mutants designed by a combination of computer-aided approach and beneficial point mutations. The specific activity (131.6 U/mg) of the best variant D1 was 4.14-fold that of the wild type, and t1/2 and T5010 increased by 13 min and 7 °C, respectively. D1 could effectively improve the solubility and emulsification of wheat proteins, more than twice the effect of the wild type. We also discussed the mechanism underlying the improved properties of D1. In summary, we not only provide a universal one-step purification and activation method to facilitate zymogen engineering but also obtain an excellent PG mutant.