In order to improve the emulsifying properties of soy protein around isoelectric point, soy protein isolate (SPI) and γ-polyglutamic acid (γ-PGA) complexes were prepared by electrostatic interaction. The formation of SPI-γ-PGA electrostatic complex and emulsifying properties were investigated by monitoring turbidity, zeta potential, intrinsic fluorophores, emulsion characterization, and microstructure observation. The results showed that the formation of SPI-γ-PGA electrostatic complex was identified through turbidimetric analysis and zeta-potential measurement. Intrinsic fluorescence spectrum indicated internal structure changes of electrostatic complexes. Furthermore, SPI-γ-PGA complex-stabilized emulsions showed better stability with small droplet sizes and slow growth as well as the uniform microstructure around the isoelectric point (pH 4.0-5.0) than SPI-formed emulsions. Under the different thermal treatments and ionic strengths, emulsions stabilized by SPI-γ-PGA-soluble complex resulted in improved emulsion stability to environmental stresses. This may be attributed to the increased steric repulsion and electrostatic repulsion by SPI-γ-PGA complexes at oil-water interfaces. The findings derived from this research would provide theoretical reference about SPI-γ-PGA electrostatic complex that can be applied in acid beverages and developed a novel plant-based sustainable stabilizer for emulsions. PRACTICAL APPLICATION: The electrostatic interaction between SPI and γ-PGA improved the emulsifying characteristics of soy protein around isoelectric point. The results derived from this research would expand applications of SPI-γ-PGA-soluble electrostatic complex that can be applied in acid beverages, as well as a novel plant-based sustainable stabilizer for emulsions.

While purifying a regular monospecific antibody, we found that the Protein A step yield was much lower than expected. Further studies revealed that the antibody formed large-size aggregates that did not bind to the Protein A resin, hence leading to dropped recovery. In an attempt to solve this low yield issue, we found that mildly acidic pH or ammonium sulfate treatment can partially convert the aggregates into monomers. In addition, when acidic pH treated culture harvest was processed by Protein A chromatography, the yield was restored to the normal range, suggesting that the monomers recovered from aggregates regained Protein A binding capability. Thus, low pH treatment of culture harvest can be potentially used as a general approach for improving Protein A step yield in cases where non-binding antibody aggregates are formed through noncovalent interactions.

The present study proposed the synthesis of low-toxicity and eco-friendly spherically shaped manganese oxides (α-MnO2, Mn2O3, and Mn3O4) by using the chemical precipitation method. The unique variable oxidation states and different structural diversity of manganese-based materials have a strong effect on fast electron transfer reactions. XRD, SEM, and BET analyses were used to confirm the structure morphology, higher surface area, and excellent porosity. The catalytic activity of as-prepared manganese oxides (MnOx) was investigated for the rhodamine B (RhB) organic pollutant with peroxymonosulfate (PMS) activation under the condition of control pH. In acidic conditions (pH = 3), complete RhB degradation and 90% total organic carbon (TOC) reduction were attained in 60 min. The effects of operating parameters such as solution pH, PMS loading, catalyst dosage, and dye concentration on RhB removal reduction were also tested. The different oxidation states of MnOx promote the oxidative-reductive reaction under acidic conditions and enhance the SO4•-/•OH radical formation during the treatment, whereas the higher surface area offers sufficient absorption sites for interaction of the catalyst with pollutants. A scavenger experiment was used to investigate the generation of more reactive species that participate in dye degradation. The effect of inorganic anions on divalent metal ions that genuinely occur in water bodies was also studied. Additionally, separation and mass analysis were used to investigate the RhB dye degradation mechanism at optimum conditions based on the intermediate\'s identification. Repeatability tests confirmed that MnOx showed superb catalytic performance on its removal trend.

Seneca Valley virus (SVV, also known as Senecavirus A), an oncolytic virus, is a nonenveloped, positive-strand RNA virus and the sole member of the genus Senecavirus within the family Picornaviridae. The mechanisms of SVV entry into cells are currently almost unknown. In the present study, we found that SVV entry into HEK293T cells is acidic pH-dependent by using ammonium chloride (NH4Cl) and chloroquine, both of which could inhibit SVV infection. We confirmed that dynamin II is required for SVV entry by using dynasore, silencing the dynamin II protein, or expressing the dominant-negative (DN) K44A mutant of dynamin II. Then, we discovered that chlorpromazine (CPZ) treatment or knockdown of the clathrin heavy chain (CLTC) protein significantly inhibited SVV infection. In addition, overexpression of CLTC promoted SVV infection. Caveolin-1 and membrane cholesterol were also required for SVV endocytosis. Notably, utilizing genistein, EIPA or nocodazole, we observed that macropinocytosis and microtubules are not involved in SVV entry. Furthermore, overexpression of the Rab7 and Rab9 proteins but not the Rab5 or Rab11 proteins promoted SVV infection. The findings were further validated by the knockdown of four Rabs and Lamp1 proteins, indicating that after internalization, SVV is transported from late endosomes to the trans-Golgi network (TGN) or lysosomes, respectively, eventually releasing its RNA into the cytosol from the lysosomes. Our findings concretely revealed SVV endocytosis mechanisms in HEK293T cells and provided an insightful theoretical foundation for further research into SVV oncolytic mechanisms.

Enhanced glycolysis is a distinct feature associated with numerous stem cells and cancer cells. However, little is known about its regulatory roles in gene expression and cell fate determination. Here, we confirm that glycolytic metabolism and lactate production decrease during the differentiation of mouse embryonic stem cells (mESCs). Importantly, acidic pH due to lactate accumulation can transiently prevent the silencing of mESC self-renewal in differentiation conditions. Furthermore, acidic pH partially blocks the differentiation of human ESCs (hESCs). Mechanistically, acidic pH downregulates AGO1 protein and de-represses a subset of mRNA targets of miR-290/302 family of microRNAs which facilitate the exit of naive pluripotency state in mESCs. Interestingly, AGO1 protein is also downregulated by acidic pH in cancer cells. Altogether, this study provides insights into the potential function and underlying mechanism of acidic pH in pluripotent stem cells (PSCs).

Microalgae have become the hotspot of recent researches as heavy metals (HMs) adsorbent and biodiesel production feedstock. In this study, the cell growth, lipid production and Cr6+ removal of Parachlorella kessleri R-3 at pH 3.5 and 15 °C were investigated. It was found that low concentration of Cr6+ (0.5 to 2 mg/L) promoted the algal growth, whereas Cr6+ higher than 5 mg/L inhibited the growth of P. kessleri R-3. Biomass concentration (2.40 g/L) and lipid productivity (131.79 mg/L d-1) reached the highest at 2 mg/L Cr6+, and lipid content (61.03 %) reached the highest at 5 mg/L Cr6+. The maximum Cr6+ removal efficiency of 91 % was obtained at 0.5 mg/L Cr6+ treatment. Furthermore, fatty acid composition analysis showed that strain R-3 had a high C16-18 content of 74.88-89.21 %. This study provides new insight into the treatment of HMs and lipid production in cold regions.

The acidic pH and total amount of SO2 are both important quality control indexes of wine, but conventional detection techniques depend heavily on specialized instrument and professional staff, thus are not available to general customers. In this paper, a hemicyanine-based colorimetric and fluorescent probe Hcy-Py was designed and synthesized. It responded to bisulfite selectively with a LOD of 0.68 μM and responded to proton with a pKa of 3.78. Upon the treatment of solutions with different pH values and concentrations of bisulfite, the probe-loaded paper strips displayed distinct color changes under both natural light and UV lamp. When a real white wine sample was subjected to the paper strip experiment, pH as well as bisulfite concentration could be determined by naked-eye quickly and conveniently, thus a visual detection of acidic pH and bisulfite in white wine without involving any sophisticated instrument or professional skill was successfully demonstrated.

Mycobacterium tuberculosis (Mtb), the pathogenic agent of tuberculosis, remains a primary inducement of morbidity and mortality globally. Mtb have evolved mechanisms to recognize diverse signals, such as acidic pH within phagolysosomes and therefore to reprogram multiple physiological and metabolic processes to adapt to intracellular survival. Moreover, lysine malonylation has been suggested to participate in regulation of enzymes in carbon metabolism. However, lysine malonylation in Mtb and its association with acidic pH associated metabolism adaptation remain unknown. Here, we systematically characterized the comparative malonylome of Mtb H37Rv grown in normal (7H9-Tyloxapol (Ty)-7.4) and acidic (7H9-Ty-4.5) medium mimicking lysosome pH. In total, 2467 lysine malonylation sites within 1026 proteins were identified, which related to diverse biological processes, particularly accumulated in metabolic process. 1090 lysine malonylation sites from 562 proteins were quantified, among which 391 lysine malonylation sites in 273 protein were down-regulated while 40 lysine malonylation sites from 36 proteins were up-regulated in acidic medium, indicating that malonylation may participate in acidic pH associated metabolism. Accordingly, the enzyme activity of GlcB was reduced under acidic stress corresponding to decreased malonylation of GlcB compared with that of normal condition and this was further demonstrated by site-specific mutations. We further found that Mtb-CobB, a sirtuin-like deacetylase and desuccinylase, involved in demalonylase activity. Together, the Mtb malonylome not only indicates the critical role of malonylation in metabolism regulation, but may provide new insights of malonylation on metabolism adaptation to acidic micro-environment in vivo.

Cotton bollworm (Helicoverpa armigera) is a worldwide agricultural pest in which the transport of pheromones is indispensable and perceived by pheromone-binding proteins (PBPs). However, three-dimensional structure, pheromone binding, and releasing mechanisms of PBPs are not completely illustrated. Here, we solved three structures of the cotton bollworm HarmPBP1 at different pH values and its complex with ligand, Z-9-hexadecenal. Although apo-HarmPBP1 adopts a common PBP scaffold of six α-helices surrounding a predominantly hydrophobic central pocket, the conformation is greatly distinct from other apo-PBPs. The Z-9-hexadecenal is bound mainly by hydrophobic interaction. The pheromone can enter this cavity through an opening between the helices α5 and α6, as well as the loop between α3 and α4. Structural analysis suggests that ligand entry into the pocket is followed by a shift of Lys94 and Lys138, which may act as a lid at the opening of the pocket. Acidic pH will cause a subtle structural change of the lid, which in turn affects its ligand-binding ability, differently from other family proteins. Taken together, this study provides structural bases for the interactions between pheromones and PBPs, the pH-induced conformational switch, and the design of small inhibitors to control cotton bollworms by disrupting male-female chemosensory communication.

Stimulus-responsive fluorescence imaging modality shows great promise for detection of tumor due to the advantages of high sensitivity, simplicity and noninvasiveness. However, some non-cancer regions including nodules and inflammation may also exhibit a stimulus-related characteristic, which cause the problem of nonspecific responsiveness and then cause \"false positive\" results for tumor recognition. Herein, hypoxia and acidic pH, two typical features strongly associated with tumor invasion, progression and metastasis in tumor microenvironment (TME), are chosen as dual stimuli to fabricate \"dual lock-and-key\" fluorescent nanoprobe for highly specific and precise imaging of tumor cells. Mesoporous silica coated gold nanorods (AuNR@mSiO2 ) are employed as nanocarrier and nanoquencher to load the pH-sensitive fluorescent reporter (Rho-TP). Azobenzene (azo) which can be reduced to amines by the highly expressed azoreductase under hypoxic conditions, is elected as the effective gatekeeper for AuNR@mSiO2 by forming complex with β-cyclodextrin polymer via host-guest interaction (azo/β-CDP). By elaborately combining the hypoxia-responsive gatekeeper and pH-responsive fluorescent signal reporter into one nanoprobe, sensitive and specific imaging of tumor cells can be realized. The fabricated dual lock-and-key fluorescent nanoprobe successfully further apply in tumor-bearing mice model, which indicate potential of early diagnosis and assessment of cancer treatment.