pH Control Research Articles

The stressostat: A novel approach in adaptive laboratory evolution to improve end-product resistance

End-product inhibition in pH-controlled batch cultures, is the major limiting factor for bacterial biomass formation in the starter culture industry as well as in many other biotechnological processes. Adaptive laboratory evolution (ALE) has emerged over the past decades as a powerful tool for phenotype optimization, but none of the existing ALE methods could select for improved end-product resistance. Therefore, we developed the stressostat (STress Resistance Evolution in Substrate Surplus) as a novel continuous ALE method. Stressostat cultivation applies end-product concentrations as constant evolutionary pressure on microorganisms in the presence of substrate surplus. In this study, we improved the lactate resistance of Lactococcus lactis FM03P in 35 days of stressostat cultivations. The lactate concentrations increased over time from 530 to 675 mM, indicating the successful selection for variants with improved lactate resistance. Thirty-four variants were isolated and grouped into four clusters based on their growth rates at high lactate concentrations. In the high-throughput screening without pH control, most isolated variants could grow at high lactate concentrations (870–928 mM), while the wild type was completely inhibited. The variants grew slower than wild type at low lactate media indicating possible evolutionary trade-off. However, in pH-controlled batch cultivations, most variants produced more biomass than the wild type. In conclusion, stressostat cultivation is a valuable method to obtain L. lactis variants with improved end-product resistance and further characterization is needed to elucidate underlying resistance mechanisms and potential industrial applications.

The simple method of lipolysis/lipogenesis balance assessment based on 14C isotope: advances and importance of pH control

The simple method of lipolysis/lipogenesis balance assessment based on 14C isotope: advances and importance of pH control

Autotrophic nitrogen polishing of secondary effluents: Alkaline pH and residual nitrate control S0-driven denitratation for downstream anammox treatment

Energy-lean nitrogen removal technologies, such as partial nitritation/anammox, often encounter effluent issues due to elevated nitrate and ammonium levels. This study proposed a novel autotrophic polishing strategy coupling sulfur-driven denitratation with anammox. To explore the denitratation potential in obtaining stable and sufficient nitrite accumulation, the effects of pH, residual nitrate level, and biomass-specific nitrate loading rate (BSNLR) were investigated in an S0-packed bed reactor at low hydraulic retention time (i.e., 0.2 h). Implementing pH and residual nitrate control strategies would be easier in practice than BSNLR control to polish secondary effluent. Alkaline pH values could realize successful nitrite accumulation without residual nitrate, and further intensify the accumulation under increased residual nitrate levels. The nitrate level was positively correlated with the nitrite accumulation efficiency. At pH 8.5 and nitrate concentration of 1.0 ± 0.8 mg N L−1, sulfur-driven denitratation could successfully maintain nitrite accumulation of 6.4 ± 1.0 mg NO2−-N L−1, ideally for the downstream anammox in case of residual ammonium levels of around 5 mg N L−1. Since Thiobacillus members play a key role in managing nitrite accumulation, their abundance should be guaranteed in the practical application.

Influence of Actinidin-Induced Hydrolysis on the Functional Properties of Milk Protein and Whey Protein Concentrates.

The main aim of the study was to establish the impact of limited proteolysis by actinidin on the functionality of selected milk protein systems. The plant protease actinidin was used to produce hydrolysates (MPHs) from milk protein concentrate (MPC) and whey protein concentrate (WPC) to 0, 5, 10 or 15% of the degree of hydrolysis (DH) at an enzyme-to-substrate ratio of 1:100 (5.21 units of actinidin activity g-1 of protein). The functionalities assessed included solubility, heat stability, emulsification and foaming properties. In general, significant changes in the functionalities of MPH were associated with the extent of hydrolysis. Solubility of hydrolysates increased with increasing %DH, with WPC showing about 97% solubility at 15% DH. Emulsifying properties were negatively affected by hydrolysis, whereas heat stability was improved in the case of WPC (~25% of heat stability increased with an increase in DH to 15%). Hydrolysates from both WPC and MPC had improved foaming properties in comparison to unhydrolysed controls. These results were also supported by changes in the FTIR spectra. Further adjustment of hydrolysis parameters, processing conditions and pH control could be a promising approach to manipulate selected functionalities of MPHs obtained using actinidin.

Autotrophic Production of the Sesquiterpene α-Humulene with Cupriavidus necator in a Controlled Bioreactor.

Cupriavidus necator is a facultative chemolithotrophic organism that grows under both heterotrophic and autotrophic conditions. It is becoming increasingly important due to its ability to convert CO2 into industrially valuable chemicals. To translate the potential of C. necator into technical applications, it is necessary to optimize and scale up production processes. A previous proof-of-principle study showed that C. necator can be used for the de novo production of the terpene α-humulene from CO2 up to concentrations of 11 mg L-1 in septum flasks. However, an increase in final product titer and space-time yield will be necessary to establish an economically viable industrial process. To ensure optimized growth and production conditions, the application of an improved process design in a gas bioreactor with the control of pH, dissolved oxygen and temperature including a controlled gas supply was investigated. In the controlled gas bioreactor, the concentration of α-humulene was improved by a factor of 6.6 and the space-time yield was improved by a factor of 13.2. These results represent an important step toward the autotrophic production of high-value chemicals from CO2. In addition, the in situ product removal of α-humulene was investigated and important indications of the critical logP value were obtained, which was in the range of 3.0-4.2.

Vanadium removal and floc characteristics of tannin biocoagulants and iron sulphate in the treatment of mine effluent

This study compared the performance of tannin (quebracho and spruce) and iron sulphate (FS) coagulants for the removal of vanadium from effluent derived from an abandoned vanadium mine. The efficacy of the materials was studied at the natural pH of the mine effluent (7.4), and at adjusted pH levels of 4 and 9. The quebracho tannin coagulant (QT) recorded impressive turbidity and vanadium removals of above 88% over a wide pH range. However, the spruce tannin coagulant (ST) was not as effective at pH 9 (39%) although it showed an improved vanadium removal of 46% and 70% at pH 7.4 and 4, respectively. This was attributed to the lower charge density of ST than that of QT, and to a loss of cationic properties at pH 9. FS performed better in terms of vanadium removal (pH 4: 86%, pH 7: 98%, pH 9: 100%) with lower coagulant doses, although effective turbidity removal required precise pH control. QT produced the largest flocs and the highest floc regrowth at pH 4 and 7. Under alkaline conditions, tannin coagulants exhibit no regrowth, which implies that a careful flocculation and floc separation step is needed. The natural organic matter fractionation results revealed that FS effectively removed hydrophilic carbon (76%) in a pH-controlled mine effluent better than tannin coagulants (QT: 12% and ST: 22%) from the pH 4 effluent; however, ST removed hydrophobic carbon entirely. All coagulants adequately removed humic substances, with FS being the most efficient (88%), followed by ST (53%) and QT (47%). X-ray photoelectron spectroscopy (XPS) analysis of jar test residues showed that some of the vanadium removed existed in the V(IV) oxidation state.

Controlling the morphology of polystyrene dumbbell particles

Controlling colloidal particles’ shape is one of the new frontiers in colloidal science. Methods that permit synthesizing polymer particles with a non-spherical shape are increasing and advancing. Of all the non-spherical particles, spheres with protrusions are one of the best investigated, and many recipes have been developed to prepare them. One of the best-developed methods, initially proposed by Park et al. (JACS, 2010), starts with linear polystyrene seed particles, which are swelled and subsequently polymerized with a mixture of styrene and 3-(trimethoxysilyl)propyl methacrylate (MPS). The latter monomer’s hybrid nature creates a hydrophilic and partially crosslinked coating. This prevents the formation of core-shell structure upon further swelling and polymerization with styrene and leads to one or more new lobes. In this work, we investigate many parameters affecting the outcome of this method, including the pH, the duration of the hydrolysis of MPS, its concentration, the presence of crosslinkers in the seed particles, and the use of a different monomer in the last step. The investigation shows the importance of pH control during MPS hydrolysis in the coating step. It also demonstrates that its concentration can effectively be used to control the morphology of the resulting particles, which range from simple one-lobed dumbbells with variable roughness to multilobed particles. Furthermore, a shorter version of the dumbbell synthesis protocol has been obtained.

Effect of Water Activity, pH, and Lactic Acid Bacteria to Inhibit Escherichia coli during Chihuahua Cheese Manufacture.

This study aimed to evaluate the effectiveness of pH control, water activity (Aw), and the addition of lactic acid bacteria (LAB) on the proliferation of Escherichia coli in the curd during the manufacturing of Chihuahua cheese. Milk proved to be an excellent culture medium for E. coli, allowing it to develop at concentrations up to 109 cfu/g. However, the presence of LAB, the pH control, Aw, and especially the use of the Cheddarization process during the Chihuahua cheese production proved to be important obstacles that inhibited the proliferation of E. coli under the conditions studied. Moreover, reducing the water activity of the curd as quickly as possible is presented as the most powerful tool to inhibit the development of E. coli during the Chihuahua cheese-making process.

Optimizing control strategies for urine nitrification: narrow pH control band enhances process stability and reduces nitrous oxide emissions

Nitrification is well-suited for urine stabilization. No base dosage is required if the pH is controlled within an appropriate operating range by urine feeding, producing an ammonium-nitrate fertilizer. However, the process is highly dependent on the selected pH set-points and is susceptible to process failures such as nitrite accumulation or the growth of acid-tolerant ammonia-oxidizing bacteria. To address the need for a robust and reliable process in decentralized applications, two different strategies were tested: operating a two-position pH controller (inflow on/off) with a narrow pH control band at 6.20/6.25 (∆pH = 0.05, narrow-pH) vs. a wider pH control band at 6.00/6.50 (∆pH = 0.50, wide-pH). These variations in pH also cause variations in the chemical speciation of ammonia and nitrite and, as shown, the microbial production of nitrite. It was hypothesized that the higher fluctuations would result in greater microbial diversity and, thus, a more robust process. The diversity of nitrifiers was higher in the wide-pH reactor, while the diversity of the entire microbiome was similar in both systems. However, the wide-pH reactor was more susceptible to tested process disturbances caused by increasing pH or temperature, decreasing dissolved oxygen, or an influent stop. In addition, with an emission factor of 0.47%, the nitrous oxide (N2O) emissions from the wide-pH reactor were twice as high as the N2O emissions from the narrow-pH reactor, most likely due to the nitrite fluctuations. Based on these results, a narrow control band is recommended for pH control in urine nitrification.

Room temperature bio-engineered multifunctional carbonates for CO2 sequestration and valorization

This contribution reports, for the first time, on an entirely green bio-engineering approach for the biosynthesis of single phase crystalline 1-D nano-scaled calcite CaCO3. This was validated using H2O as the universal solvent and natural extract of Hyphaene thebaica fruit as an effective chelating agent. In this room temperature green process, CaCl2 and CO2 are used as the unique source of Ca and CO3 respectively in view of forming nano-scaled CaCO3 with a significant shape anisotropy and an elevated surface to volume ratio. In terms of novelty, and relatively to the reported scientific and patented literature in relation to the fabrication of CaCO3 by green nano-chemistry, the current cost effective room temperature green process can be singled out as per the following specificities: only water as universal solvent is used, No additional base or acid chemicals for pH control, No additional catalyst, No critical or supercritical CO2 usage conditions, Only natural extract of thebaica as a green effective chelating agent through its phytochemicals and proper enzematic compounds, room Temperature processing, atmospheric pressure processing, Nanoscaled size particles, and Nanoparticles with a significant shape anisotropy (1-D like nanoparticles). Beyond and in addition to the validation of the 1-D synthesis aspect, the bio-engineered CaCO3 exhibited a wide-ranging functionalities in terms of highly reflecting pigment, an effective nanofertilizer as well as a potential binder in cement industry.

Efficient Production of Chlorogenic Acid in Escherichia coli Via Modular Pathway and Cofactor Engineering.

Chlorogenic acid is a natural phenolic compound widely used in the food and daily chemical industries. Compared to plant extraction, microbial cell factories provide a green and sustainable production method for the production of chlorogenic acid. However, complex metabolic flux distribution and potential byproducts limited the biosynthesis of chlorogenic acid in microorganisms. A de novo biosynthesis pathway for chlorogenic acid was constructed in Escherichia coli via modular engineering. Increasing the shikimate pathway flux greatly promoted chlorogenic acid production, and the influence of pyruvate metabolism on chlorogenic acid synthesis was also explored. The supply of cofactors for the key enzymes quinate/shikimate 5-dehydrogenase (YdiB) and 4-hydroxyphenylacetate 3-monooxygenase (HpaBC) was enhanced by a cofactor regeneration system. Furthermore, mutants of YdiB were verified for chlorogenic acid production in vivo. Chlorogenic acid browning occurred when the buffer pH of the buffer exceeded 6.0, but two-stage pH control achieved a chlorogenic acid titer of 2789.2 mg/L in a 5 L fermenter, the highest reported to date. This study provided a strategy for the efficient production of chlorogenic acid from simple carbon sources.

Postbiotics Production of Candidate-Probiotic Lactiplantibacillus plantarum AC131 with Renewable Bio Resources.

Lactiplantibacillus plantarum is a versatile specie, well known as a producer of lactic acid (LA) and other metabolites with biotechnological significance. The present work characterizes growth and lactic acid production of the candidate-probiotic strain L. plantarum AC131, from Bulgarian white brined cheeses. Different nutritional media with ingredients from renewable resources-reduced sugars from dried distillers' grains with soluble (DDGS) and waste waters from the water-vapor distillation of Bulgarian Rosa alba L. and Rosa damascena Mill. essential oil-were assessed. The results obtained showed significant LA production (up to 95% conversion) in modified MRS broth with reducing sugars from DDGS hydrolysates. The addition of R. alba L. and R. damascena Mill. distillation effluents stimulated the growth and biological activity of postbiotics produced by L. plantarum AC131. In both experimental approaches, a statistically significant inhibition (from 20 to 60%) of E. coli HB 101 growth was found during 24 h exposure and a variable effect on the biofilm formed. In conclusion, reducing sugars from DDGS hydrolysates can be successfully used as a carbon source for lactic acid production. In the case of fermentation without pH control, the process is product inhibited, while with pH control, nearly full conversion was achieved. Postbiotics produced during the process of fermentation showed a variety of biological activity and inhibitory effects on the growth of Escherichia coli HB 101.

Multi-responsive Electropolymer Surface Coatings Based on Azo Molecular Switches and Carbazoles: Light, pH, and Electrochemical Control of Z→E Isomerization in Thin Films.

Light-responsive surfaces are attracting increasing interest, not least because their physicochemical properties can be selectively and temporally controlled by a non-invasive stimulus. Most existing immobilization strategies involve the chemical attachment of light-responsive moieties to the surface, although this approach often suffers from a low surface concentration of active species or a high inhomogeneity of applied coatings. Herein, electropolymerization of carbazoles as a facile and rapid approach for preparing light-responsive azo-based surface coatings is presented. The electrochemical oxidative polymerization of bis-carbazole containing azo-monomers yields stable films, in which the photochemical properties and specific pH sensitivity of azo molecular switches are retained. Moreover, the molecular design enables electrocatalytic control over Z→E azo double bond isomerization facilitated by the conductive polycarbazole backbone. Ultimately, the high degree of control over macromolecular properties yields conductive surface coatings responsive to a range of stimuli, showing great promise as a strategy for versatile application in organic electronics.

Elemental sulfur autotrophic partial denitrification (S0-PDN) with high pH and free ammonia control strategy for low-carbon wastewater: From performance to microbial mechanism

Elemental sulfur autotrophic partial denitrification (S0-PDN) with high pH and free ammonia control strategy for low-carbon wastewater: From performance to microbial mechanism

Aqueous metal ion leaching processes from high sulfur coal fly ash for carbon mineralization: The importance of pH control on cation extraction, carbonate purity, and silicon Q structure

Aqueous metal ion leaching processes from high sulfur coal fly ash for carbon mineralization: The importance of pH control on cation extraction, carbonate purity, and silicon Q structure

Disinfection Effects and Disinfection by-products Generation Characteristics of a Continuous Hypochlorous Acid Water Producing Device Using Citric Acid as a pH Control Agent

The use of disinfectants and disinfecting devices is becoming increasingly common in homes as they prevent the development and spread of pathogens while improving hygiene. Common disinfectants include chlorine compounds, alcohol, and quaternary ammonium compounds. Among them, chlorine compounds are easily manufactured and commonly used in the form of sodium hypochlorite. When chlorine compounds are dissolved in water, free chlorine divides into hypochlorous acid and hypochlorite ions with different molar fractions depending on solution pHs. Maintaining a high hypochlorous acid fraction by adjusting the solution pH is crucial as hypochlorous acid is more than 100 times more effective as a disinfectant than hypochlorite ions. However, due to the chemical nature of its conversion to hypochlorite ions, hypochlorous acid is not very stable, making it more efficient to prepare and consume it at the time of use. In this study, we used citric acid as a pH adjuster to determine the optimal conditions required for producing electrolyzed water. We built a device capable of continuously producing electrolyzed water containing hypochlorous acid, and we analyzed the efficiency of the produced electrolyzed water using disinfection tests and measurements of disinfection by-products.

Significant Variability in the Developmental Toxicity of Representative Perfluoroalkyl Acids as a Function of Chemical Speciation.

Current toxicological data of perfluoroalkyl acids (PFAAs) are disparate under similar exposure scenarios. To find the cause of the conflicting data, this study examined the influence of chemical speciation on the toxicity of representative PFAAs, including perfluorooctanoic acid (PFOA), perfluorobutane carboxylic acid (PFBA), and perfluorobutanesulfonic acid (PFBS). Zebrafish embryos were acutely exposed to PFAA, PFAA salt, and a pH-negative control, after which the developmental impairment and mechanisms were explored. The results showed that PFAAs were generally more toxic than the corresponding pH control, indicating that the embryonic toxicity of PFAAs was mainly caused by the pollutants themselves. In contrast to the high toxicity of PFAAs, PFAA salts only exhibited mild hazards to zebrafish embryos. Fingerprinting the changes along the thyroidal axis demonstrated distinct modes of endocrine disruption for PFAAs and PFAA salts. Furthermore, biolayer interferometry monitoring found that PFOA and PFBS acids bound more strongly with albumin proteins than did their salts. Accordingly, the acid of PFAAs accumulated significantly higher concentrations than their salt counterparts. The present findings highlight the importance of chemical forms to the outcome of developmental toxicity, calling for the discriminative risk assessment and management of PFAAs and salts.

Verifying the Optimal Extraction Method and Content of Red Okra Extracts and Crude Pectin Using Response Surface Methodology (RSM)

Recently, many studies have reported the properties and functionality of okra pectin. However, these studies are about green okra pods, and pectin in red okra pods, stems, and roots has not been reported. Therefore, this study aimed to optimize the red okra extraction method using response surface methodology (RSM) analysis and evaluate the effects of extraction time, temperature, solvent ratio, and pH on the extract yield and crude pectin content. Based on RSM analysis, 4.35 h, 98.04 °C, 23.34 solvent ratio, and pH 3.36 are the optimal parameters for extracting crude pectin from red okra, and the crude pectin content was predicted to be 40.83%. When red okra was extracted under these extraction conditions (4 h, 100 °C, 23 solvent ratio, and pH 3), the extraction yield was 45.26%, and the crude pectin content was 38.42%, which was similar to the yield obtained under the conditions derived from the RSM analysis. In addition, the pH control using hydrochloric acid was replaced with citric acid, and the changes in extract yield and crude pectin content were compared. When citric acid was used, the extract yield was 49.15% (8.6% increase), and the crude pectin content was 42.76 ± 2.56% (11.3% increase); compared to when hydrochloric acid was used, the yield increased. Finally, the standardization of red okra raw materials was determined by analyzing the extraction yield and crude pectin content by part, harvest time, and size of red okra using the established extraction method using citric acid. As a result, it was confirmed that the extraction yield obtained from the established extraction method was reached from the extraction of red okra fruits up to 12 cm in size that were harvested between July and November. Additionally, compared to the non-pectin fraction, the crude pectin fraction isolated from red okra pod extract showed significantly higher total phenolic content (TPC) and total flavonoid content (TFC). These findings, reported for the first time, may contribute to the development of processes to purify red okra pectin, functional evaluation studies of pectin, and potential applications of red okra extract in various industries and research.

The Impact of Trisodium Phosphate on Chloride-Driven Under Deposit Corrosion in Steam Generators

Carbon or low alloy steel tubes in steam generators (or boilers) are potentially vulnerable to under deposit corrosion (UDC), arising from the formation of porous magnetite deposits on the waterside heat-transfer surfaces. Beneath these deposits, “wick-boiling” causes a concentration of contaminants (such as chlorides), which eventually leads to rapid corrosion. In this work, the corrosion of carbon steel has been investigated in hot acid chloride solutions that simulate the concentrated local environments formed during UDC. Trisodium phosphate (TSP) is sometimes dosed into boilers for pH control. This work has shown that TSP addition such that the phosphate concentration equals the chloride concentration dramatically reduces the corrosion rate in these simulated environments from >20 mm/y to <0.1 mm/y. Additionally, a model of wick-boiling beneath deposits has been used to analyze the concentration of chlorides and phosphates during the initiation stages of UDC, suggesting that dosing of only 100 ppb of TSP into bulk boiler water should be sufficient to increase the critical deposit thickness (required for UDC) by >100 µm across a wide range of operational scenarios.

Removal of heavy metals from mine tailings by in-situ bioleaching coupled to electrokinetics

This work utilizes a combined biological-electrochemical technique for the in-situ removal of metals from polluted mine tailings. As the main novelty point it is proposed to use electrokinetics (EK) for the in-situ activation of a bioleaching mechanism into the tailings, in order to promote biological dissolution of metal sulphides (Step 1), and for the subsequent removal of leached metals by EK transport out of the tailings (Step 2). Mine tailings were collected from an abandoned Pb/Zn mine located in central-southern Spain. EK-bioleaching experiments were performed under batch mode using a lab scale EK cell. A mixed microbial culture of autochthonous acidophilic bacteria grown from the tailings was used. Direct current with polarity reversal vs alternate current was evaluated in Step 1. In turn, different biological strategies were used: biostimulation, bioaugmentation and the abiotic reference test (EK alone). It was observed that bioleaching activation was very low during Step 1, because it was difficult to maintain acidic pH in the whole soil, but then it worked correctly during Step 2. It was confirmed that microorganisms successfully contributed to the in-situ solubilization of the metal sulphides as final metal removal rates were improved compared to the conventional abiotic EK (best increases of around 40% for Cu, 162% for Pb, 18% for Zn, 13% for Mn, 40% for Ni and 15% for Cr). Alternate current seemed to be the best option. The tailings concentrations of Fe, Al, Cu, Mn, Ni and Pb after treatment comply with regulations, but Pb, Cd and Zn concentrations exceed the maximum values. From the data obtained in this work it has been observed that EK-bioleaching could be feasible, but some upgrades and future work must be done in order to optimize experimental conditions, especially the control of soil pH in acidic values.