Preferred Carbon Research Articles

Microalgae-based constructed wetland system enhances nitrogen removal and reduce carbon emissions: Performance and mechanisms

Combination of constructed wetlands (CWs) and microalgae-based technologies has been proved as effective wastewater treatment option; however, little attention was paid to investigate the optimal combination ways. This study showed that the integrated system (IS) connecting microalgal pond with CWs exhibited improved pollutant-removal efficiencies and preferred carbon reduction effects compared to other alternatives such as coupled system or independent CWs. Microbial analysis demonstrated that core microorganisms (e.g., Acinetobacter and Thermomonas) of the IS were mostly associated with carbon, nitrogen, and energy metabolism. Based on co-occurrence networks, microbial quantity with denitrification function in the IS accounted for 71.01 % of the microorganism related to nitrogen metabolism, which was higher than that of 48.84 % in the independent CWs, indicating that the presence of microalgae in IS played important role in promoting biological denitrification. These findings provide insights into the microbial mechanism and highlights the complementary effects between microalgae and CWs.

LipR functions as an intracellular pH regulator in Bacillus thuringiensis under glucose conditions.

Intracellular pH critically affects various biological processes, and an appropriate cytoplasmic pH is essential for ensuring bacterial growth. Glucose is the preferred carbon source for most heterotrophs; however, excess glucose often causes the accumulation of acidic metabolites, lowering the intracellular pH and inhibiting bacterial growth. Bacillus thuringiensis can effectively cope with glucose-induced stress; unfortunately, little is known about the regulators involved in this process. Here, we document that the target of the dual-function sRNA YhfH, the lipR gene, encodes a LacI-family transcription factor LipR as an intracellular pH regulator when B. thuringiensis BMB171 is suddenly exposed to glucose. Under glucose conditions, lipR deletion leads to early growth arrest by causing a rapid decrease in intracellular pH (~5.4). Then, the direct targets and a binding motif (GAWAWCRWTWTCAT) of LipR were identified based on the electrophoretic mobility shift assay,the DNase-I footprinting assay, and RNA sequencing, and the gapN gene encoding a key enzyme in glycolysis was directly inhibited by LipR. Furthermore, Ni2+ is considered a possible effector for LipR. In addition to YhfH, the lipR expression was coregulated by itself, CcpA, and AbrB. Our study reveals that LipR plays a balancing role between glucose metabolism and intracellular pH in B. thuringiensis subjected to glucose stress.

Iron effects localization of Ght5 in fission yeast.

Iron is an essential cofactor for eukaryotic cells, as well as a toxic metal under certain conditions. On the other hand, glucose is the preferred energy and carbon source by most organisms and is an important signaling molecule in the regulation of biological processes. In Schizosaccharomyces pombe, the Ght5 hexose transporter, known as a high affinity glucose transporter, is required for cell proliferation in the low glucose condition. In this study, it is aimed to investigate the effects of iron stress on the Ght5 hexose transporter under glucose repression and derepression conditions. The effect of iron stress on the expression profile of the ght5 gene was analyzed by RT-qPCR and western blot. The localization of the Ght5-mNeongreen fusion protein examined with confocal microscopy. As a result, it was revealed that iron stress had an inhibitory effect on ght5 expression, and it altered Ght5 localization on the cell surface causing accumulation in the cytoplasm.

An atomic/molecular-level strategy for the design of a preferred nitrogen-doped carbon nanotube cathode for Li-O2 batteries

The formation and growth of Li2O2, the primary discharge product with extremely low conductivity, will cause the passivation of the cathode and the failure or even death of Li-O2 batteries. In this paper, by means of molecular dynamics and density functional theory, we systematically investigated the morphology evolution and electron transport pathway of (Li2O2)n on the surface of nitrogen-doped carbon nanotubes (NCNTs) and the effects of the possible N configuration on the growth stability of (Li2O2)n. Our results demonstrated that the morphology of (Li2O2)n on the surface of NCNTs undergos a process from scattered small-sized clusters to film-like (axial) and toroidal (circumferential) large clusters, and finally completely surrounded the NCNTs. Compared with graphitic N1 and graphitic N2, Li2O2 tends to aggregate stably on the surface of pyridinic N3 and pyridinic N4, which can effectively reduce the capacity loss caused by Li2O2 branches and free state Li2O2. In addition, a stable gap exists between Li2O2 and NCNTs, which is attributed to the Coulomb force and van der Waals interaction. Importantly, (Li2O2)n with p-type semiconductor characteristics acts as a bridge between NCNTs, and electrons are mainly transferred along the surface of (Li2O2)n clusters. Moreover, increasing the wall number of NCNTs and using NCNT bundles were found to be beneficial to improving the deposition stability of (Li2O2)n. Notably, pyrrolic N3 with unstable structure is not recommended for practical application. These results offer general guidelines for designing highly active catalysts for Li-O2 batteries in terms of the optimization of the NCNT-based cathode, and the unique research methods also shed more light on the analysis of the discharge product behavior of other potential metal-air batteries.

A Manganese-independent Aldolase Enables Staphylococcus aureus To Resist Host-imposed Metal Starvation.

The preferred carbon source of Staphylococcus aureus and many other pathogens is glucose, and its consumption is critical during infection. However, glucose utilization increases the cellular demand for manganese, a nutrient sequestered by the host as a defense against invading pathogens. Therefore, bacteria must balance glucose metabolism with the increasing demand that metal-dependent processes, such as glycolysis, impose upon the cell. A critical regulator that enables S. aureus to resist nutritional immunity is the ArlRS two-component system. This work revealed that ArlRS regulates the expression of FdaB, a metal-independent fructose 1,6-bisphosphate aldolase. Further investigation revealed that when S. aureus is metal-starved by the host, FdaB functionally replaces the metal-dependent isozyme FbaA, thereby allowing S. aureus to resist host-imposed metal starvation in culture. Although metal-dependent aldolases are canonically zinc-dependent, this work uncovered that FbaA requires manganese for activity and that FdaB protects S. aureus from manganese starvation. Both FbaA and FdaB contribute to the ability of S. aureus to cause invasive disease in wild-type mice. However, the virulence defect of a strain lacking FdaB was reversed in calprotectin-deficient mice, which have defects in manganese sequestration, indicating that this isozyme contributes to the ability of this pathogen to overcome manganese limitation during infection. Cumulatively, these observations suggest that the expression of the metal-independent aldolase FdaB allows S. aureus to alleviate the increased demand for manganese that glucose consumption imposes, and highlights the cofactor flexibility of even established metalloenzyme families. IMPORTANCE Staphylococcus aureus and other pathogens consume glucose during infection. Glucose utilization increases the demand for transition metals, such as manganese, a nutrient that the host limits as a defense mechanism against invading pathogens. Therefore, pathogenic bacteria must balance glucose and manganese requirements during infection. The two-component system ArlRS is an important regulator that allows S. aureus to adapt to both glucose and manganese starvation. Among the genes regulated by ArlRS is the metal-independent fructose 1,6-bisphosphate aldolase fdaB, which functionally substitutes for the metal-dependent isoenzyme FbaA and enables S. aureus to survive host-imposed manganese starvation. Unexpectedly, and differing from most characterized metal-dependent aldolases, FbaA requires manganese for activity. Cumulatively, these findings reveal a new mechanism for overcoming nutritional immunity as well as the cofactor plasticity of even well-characterized metalloenzyme families.

Techno-economic comparative analysis of solar thermal collectors and high-temperature heat pumps for industrial steam generation

Industrial heat production is responsible for around 20% of total greenhouse gas emissions in Europe. To achieve the climate change goals defined in the Paris Climate Agreement, the EU commission has shifted its focus on sustainable means to generate heating. Moreover, global dependencies are leading to a re-organization of natural gas supplies. Therefore, there is a need for less vulnerable and less price-volatile solutions for heating. This paper focuses on two decarbonization technologies for industrial process heat supply: a) electricity-driven steam-generating high-temperature heat pumps (HTHP), a technology that is more efficient than fossil fuel boilers in generating steam, and b) solar parabolic trough collector (PTC), which can produce heat economically and at a minimal carbon footprint compared to other technologies. The main aim of this paper is to evaluate the levelized cost of heat (LCOH) of these technologies to fulfill a comparative techno-economic analysis. A maximum PTC collector's solar fraction limit (SFlimit) is defined to indicate when the LCOH for these two technologies is equal. This allows for distinguishing between the economic stronghold of each technology. The evaluation is carried out through the annual energy simulations using TRNSYS and Excel spreadsheets for HTHPs, while TRNSED and OCTAVE are used for the solar thermal part. Boundary conditions for European geographical constraints have been applied to establish use cases for the analysis. The result shows that the design of a PTC system with optimal SF can reach cost parity with HTHP for most of the analyzed locations. The developed methodology serves as a valuable guide to quickly determine a preferred lower carbon heat solution, thus easing the decision-making for industries.

Identification of Lactobacillus strains from human mother milk and cottage cheese revealed potential probiotic properties with enzymatic activity

The main attempt of this study is to isolate, determine potential probiotic properties and enzyme production of some lactic acid bacteria (LAB). Among all isolates, two LAB strains isolated from human mother milk and cottage cheese revealed antimicrobial activity against some tested pathogenic strains. Both isolates inhibited all the tested pathogens except Escherichia coli. The two isolates were identified by morphological, biochemical properties and then by 16S rRNA gene sequencing technique as Lactobacillus acidophilus SAM1 and Lactiplantibacillus plantarum SAM2. Potential probiotic characters were investigated. Both strains survived in relatively low pH and high bile concentrations and were able to grow at 0.5% of pancreatin concentrations. Their growth decreased by increasing phenol from 0.2% till 0.5%. Both strains did not show hemolytic activity. Coaggregation potential was exhibited by the two strains against Staphylococcus aureus and Candida albicans. Hydrophobicity of Lactobacillus acidophilus SAM1 and Lactiplantibacillus plantarum SAM2, with ethyl acetate; were 88.1% and 82.8%, respectively. Lactobacillus acidophilus SAM1 was susceptible to Ampicillin, Penicillin, Erythromycin, Ciprofloxacin and Tetracycline; on the contrary, it resists Vancomycin and Cefoxitin; while Lactiplantibacillus plantarum SAM2 resists all tested antibiotics. Maximum growth was achieved using glucose as a carbon source and yeast extract as nitrogen source for both strains; however, glucose is the most preferred carbon source for microorganisms and it prevents the uptake of carbon from other sources like yeast by catabolite repression mechanism. Lactobacillus acidophilus SAM1 produces lipase enzyme, while Lactiplantibacillus plantarum SAM2 produces amylase and protease.

The acetoin assimilation pathway of Pseudomonas putida KT2440 is regulated by overlapping global regulatory elements that respond to nutritional cues.

Many microorganisms produce and excrete acetoin (3-hydroxy-2-butanone) when growing in environments that contain glucose or other fermentable carbon sources. This excreted compound can then be assimilated by other bacterial species such as pseudomonads. This work shows that acetoin is not a preferred carbon source of Pseudomonas putida, and that the induction of genes required for its assimilation is down-modulated by different, independent, global regulatory systems when succinate, glucose or components of the LB medium are also present. The expression of the acetoin degradation genes was found to rely on the RpoN alternative sigma factor and to be modulated by the Crc/Hfq, Cyo and PTSNtr regulatory elements, with the impact of the latter three varying according to the carbon source present in addition to acetoin. Pyruvate, a poor carbon source for P. putida, did not repress acetoin assimilation. Indeed, the presence of acetoin significantly improved growth on pyruvate, revealing these compounds to have a synergistic effect. This would provide a clear competitive advantage to P. putida when growing in environments in which all the preferred carbon sources have been depleted and pyruvate and acetoin remain as leftovers from the fermentation of sugars by other microorganisms.

Regulation of hierarchical carbon substrate utilization, nitrogen fixation, and root colonization by the Hfq/Crc/CrcZY genes in Pseudomonas stutzeri.

Bacteria of the genus Pseudomonas consume preferred carbon substrates in nearly reverse order to that of enterobacteria, and this process is controlled by RNA-binding translational repressors and regulatory ncRNA antagonists. However, their roles in microbe-plant interactions and the underlying mechanisms remain uncertain. Here we show that root-associated diazotrophic Pseudomonas stutzeri A1501 preferentially catabolizes succinate, followed by the less favorable substrate citrate, and ultimately glucose. Furthermore, the Hfq/Crc/CrcZY regulatory system orchestrates this preference and contributes to optimal nitrogenase activity and efficient root colonization. Hfq has a central role in this regulatory network through different mechanisms of action, including repressing the translation of substrate-specific catabolic genes, activating the nitrogenase gene nifH posttranscriptionally, and exerting a positive effect on the transcription of an exopolysaccharide gene cluster. Our results illustrate an Hfq-mediated mechanism linking carbon metabolism to nitrogen fixation and root colonization, which may confer rhizobacteria competitive advantages in rhizosphere environments.

Effect of pH Change on the Microalgae-Based Biogas Upgrading Process

An alternative way to remove CO2 from biogas is the use of photosynthetic microorganisms, such as microalgae. This can be achieved by the operation of an open photobioreactor, connected with a mass transfer column, such as a counterflow column. This technology provides up-graded biogas with high quality. The microalgal uptake of CO2 from the biogas in counterflow columns generates pH changes in microalgae culture. To clarify the potential effect of these dynamic pH conditions in the culture, the effect of pH change on the photosynthetic activity and PSII quantum yield was studied for microalgae Chlorella sorokiniana. Thus, assays were carried out, where the pH drop reported in the counterflow columns was replicated in batch microalgae culture through HCl addition and CO2 injection, moving the culture pH from 7.0 to 5.0 and from 7.0 to 5.8, respectively. Moreover, the effect of light/darkness on photosynthetic activity was tested when the pH decreased. The results obtained in this research showed that the photosynthetic activity decreased for the light conditions when the pH was shifted by HCl addition and CO2 injection. Despite this, the value of the PSII quantum yield remained at 0.6–0.7, which means that the microalgae culture did not suffer a negative effect on the photosynthetic system of cells because a high value of PSII efficiency remained. In the same way, the results indicated that when the pH change was corrected, the photosynthetic activity recovered. Moreover, the apparent affinity constant for dissolved inorganic carbon (KDIC) was 0.9 µM at pH 5 and 112.0 µM at pH 7, which suggests that the preferred carbon source for C.sorokniana is CO2. Finally, all the results obtained indicated that the pH drop in the counter-flow column for biogas upgrading did not cause permanent damage to the photosynthetic system, and the decrease in the photosynthetic activity as a result of the pH drop can be recovered when the pH is corrected.

Insight into Na+ assistant anaerobic fermentation of waste activated sludge from carbon migration, bio-transformation and recovery perspectives

Short-chain fatty acids (SCFAs) produced from anaerobic fermentation of waste activated sludge (WAS) have been regarded as the preferred carbon source, however the carbon migration pattern has rarely been reported. This work elucidated that the Na+ assistant anaerobic fermentation could facilitate carbon source release and bio-transformation towards carbon recovery from WAS. Obvious disintegration of sludge flocs and property variation of solid–liquid interface were triggered with Na+-regulation, leading to extracellular polymeric substance disruption and microbial cell apoptosis by protein structure loosening and osmotic pressure stress. Thereby, extracellular and intracellular carbon source (e.g. proteins, carbohydrates) were simultaneously solubilized, contributing to considerable carbon migration from sludge solid into liquid phase (up to 20.43 %). Meanwhile, 61.30 % of the released carbon source was bio-transformed with the improved sludge acidification in 4-day Na+ assistant anaerobic fermentation. Substantial SCFAs of 61.39 mg C/g VSS was produced, which was mainly composed by acetate and propionate (69.07 %). As such, the overall carbon recovery rate of 12.51 % was achievable. The carbon source bio-availability in fermentative liquid indeed created economic benefits and carbon-emission reduction depending on alternative carbon source utilization and residual carbon source reduction in fermented sludge disposal.

One-Step Regio- and Stereoselective Electrochemical Synthesis of Orexin Receptor Antagonist Oxidative Metabolites.

Synthesis of drug metabolites, which often have complex structures, is an integral step in the evaluation of drug candidate metabolism, pharmacokinetic (PK) properties, and safety profiles. Frequently, such synthetic endeavors entail arduous, multiple-step de novo synthetic routes. Herein, we present the one-step Shono-type electrochemical synthesis of milligrams of chiral α-hydroxyl amide metabolites of two orexin receptor antagonists, MK-8133 and MK-6096, as revealed by a small-scale (pico- to nano-mole level) reaction screening using a lab-built online electrochemistry (EC)/mass spectrometry (MS) (EC/MS) platform. The electrochemical oxidation of MK-8133 and MK-6096 was conducted in aqueous media and found to produce the corresponding α-piperidinols with exclusive regio- and stereoselectivity, as confirmed by high-resolution nuclear magnetic resonance (NMR) characterization of products. Based on density functional theory (DFT) calculations, the exceptional regio- and stereoselectivity for this electrochemical oxidation are governed by more favorable energetics of the transition state, leading to the preferred secondary carbon radical α to the amide group and subsequent steric hindrance associated with the U-shaped conformation of the cation derived from the secondary α-carbon radical, respectively.

Redundancy in Citrate and cis-Aconitate Transport in Pseudomonas aeruginosa.

Tricarboxylates such as citrate are the preferred carbon sources for Pseudomonas aeruginosa, an opportunistic pathogen that causes chronic human infections. However, the membrane transport process for the tricarboxylic acid cycle intermediates citrate and cis-aconitate is poorly characterized. Transport is thought to be controlled by the TctDE two-component system, which mediates transcription of the putative major transporter OpdH. Here, we search for previously unidentified transporters of citrate and cis-aconitate using both protein homology and RNA sequencing approaches. We uncover new transporters and show that OpdH is not the major citrate importer; instead, citrate transport primarily relies on the tripartite TctCBA system, which is encoded in the opdH operon. Deletion of tctA causes a growth lag on citrate and loss of growth on cis-aconitate. Combinatorial deletion of newly discovered transporters can fully block citrate utilization. We then characterize transcriptional control of the opdH operon in tctDE mutants and show that loss of tctD blocks citrate utilization due to an inability to express opdH-tctCBA. However, tctE and tctDE mutants evolve heritable adaptations that restore growth on citrate as the sole carbon source. IMPORTANCE Pseudomonas aeruginosa is a bacterium that infects hospitalized patients and is often highly resistant to antibiotic treatment. It preferentially uses small organic acids called tricarboxylates rather than sugars as a source of carbon for growth. The transport of many of these molecules from outside the cell to the interior occurs through unknown channels. Here, we examined how the tricarboxylates citrate and cis-aconitate are transported in P. aeruginosa. We then sought to understand how production of proteins that permit citrate and cis-aconitate transport is regulated by a signaling system called TctDE. We identified new transporters for these molecules, clarified the function of a known transport system, and directly tied transporter expression to the presence of an intact TctDE system.

Growth ability, carbon source utilization and biochemical features of the new specie Zalaria obscura

This research investigated the characteristics of Zalaria obscura LS31012019 in terms of growth ability in different media (SDB, YPD and TSB) and temperatures (22, 25 and 37 °C), utilization of several carbon sources (Glucose, Fructose, Lactose, Sucrose, Xylose, Glycerol and Mannitol at 5, 2 and 1%) and several biochemical features (total protein content, Glutathione, pigments), in comparison with those of the phylogenetically related Aureobasidium pullulans ATCC 15233. The best growth of Z. obscura LS31012019 was obtained in YPD at 25 °C with the highest OD value (0.45) after 144 h of incubation, similar to that of A. pullulans ATCC 15233 (0.48). Glucose resulted the preferred carbon source for both the considered yeasts but also sucrose resulted in efficacy supporting the growth of Z. obscura LS31012019 and A. pullulans ATCC 15233, for their ability in converting sucrose to glucose and fructose and the latter into glucose. Interestingly, Z. obscura LS31012019 utilized also glycerol and mannitol. The biochemical analysis showed the similarity of protein profile in Z. obscura LS31012019 and A. pullulans ATCC 15233 (from 90 to 20 kDa) and a reduced GSH content (0.321 and 0.233 µmol/mg). The pigments extraction with hexane generated a yellow oleaginous pellet in both the strains, while a yellow solid matrix more intensely coloured in A. pullulans ATTC 15233 was visible with the following solvent extractions. Overall, our data showed that Z. obscura LS31012019 can grow in different media and temperatures and utilize carbon sources apart from glucose and sucrose, shifting to a non-fermentative metabolism. These results improve the information regarding the characteristics of Z. obscura, opening a new field of investigation for the possible application of new species of black yeasts in human application.

Elevating Phospholipids Production Yarrowia lipolytica from Crude Glycerol.

Phospholipids (PLs) are a class of lipids with many proven biological functions. They are commonly used in lipid replacement therapy to enrich cell membranes damaged in chronic neurodegenerative diseases, cancer, or aging processes. Due to their amphipathic nature, PLs have been widely used in food, cosmetic, and pharmaceutical products as natural emulsifiers and components of liposomes. In Yarrowia lipolytica, PLs are synthesized through a similar pathway like in higher eukaryotes. However, PL biosynthesis in this yeast is still poorly understood. The key intermediate in this pathway is phosphatidic acid, which in Y. lipolytica is mostly directed to the production of triacylglycerols and, in a lower amount, to PL. This study aimed to deliver a strain with improved PL production, with a particular emphasis on increased biosynthesis of phosphatidylcholine (PC). Several genetic modifications were performed: overexpression of genes from PL biosynthesis pathways as well as the deletion of genes responsible for PL degradation. The best performing strain (overexpressing CDP-diacylglycerol synthase (CDS) and phospholipid methyltransferase (OPI3)) reached 360% of PL improvement compared to the wild-type strain in glucose-based medium. With the substitution of glucose by glycerol, a preferred carbon source by Y. lipolytica, an almost 280% improvement of PL was obtained by transformant overexpressing CDS, OPI3, diacylglycerol kinase (DGK1), and glycerol kinase (GUT1) in comparison to the wild-type strain. To further increase the amount of PL, the optimization of culture conditions, followed by the upscaling to a 2 L bioreactor, were performed. Crude glycerol, being a cheap and renewable substrate, was used to reduce the costs of PL production. In this process 653.7 mg/L of PL, including 352.6 mg/L of PC, was obtained. This study proved that Y. lipolytica is an excellent potential producer of phospholipids, especially from waste substrates.

The TonB-dependent uptake of pyrroloquinoline-quinone (PQQ) and secretion of gluconate by Escherichia coli K-12.

Glucose is taken up by Escherichia coli through the phosphotransferase system (PTS) as the preferred carbon source. PTS mutants grow with glucose as a carbon source only in the presence of pyrroloquinoline quinone (PQQ), which is needed as a redox cofactor for the glucose dehydrogenase Gcd. The membrane-anchored Gcd enzyme oxidises glucose to gluconolactone in the periplasm. For this reaction to occur, external supply of PQQ is required as E. coli is unable to produce PQQ de novo. Growth experiments show that PqqU (previously YncD) is the TonB-ExbBD-dependent transporter for PQQ through the outer membrane. PQQ protected the cells from the PqqU-dependent phage IsaakIselin (Bas10) by competition for the receptor protein. As a high affinity uptake system, PqqU allows E. coli to activate Gcd even at surrounding PQQ concentrations of about 1nmoL/L. At about 30-fold higher PQQ concentrations, the activation of Gcd gets PqqU independent. Due to its small size, Pqq may also pass the outer membrane through porins. The PQQ-dependent production of gluconate has been demonstrated in many plant growth-promoting bacteria that solubilise phosphate minerals in the soil by secreting this acid. Under phosphate limiting conditions also E. coli induces the glucose dehydrogenase and secretes gluconate, even in absence of PTS, that is, even when the bacterium is unable to grow on glucose without PQQ.

Evaluating the efficacy of bacterial consortium for decolorization of diazo dye mixture.

Textile wastewater contains dyes mixed with other contaminants in various concentrations. Bacteria-mediated decolorization and degradation of azo dyes have achieved momentum as a method of treatment attributed to their inexpensive, eco-friendly, and application to a wide range of azodyes. However, a single species of bacteria is inefficient in decolorizing diverse groups of dyeswhich is one of the most significant challenges for environmental technologists working in bioremediation. In the present study, an aerobic bacterial consortium AUJ consisting of six different bacterial strains (Pseudomonas stutzeri AK1, Pseudomonas stutzeri AK2, Pseudomonas stutzeri AK3, Bacillus spp. AK4, Pseudomonas stutzeri AK5, and Pseudomonas stutzeri AK6)removed the individual azo dyes in the 24-94% range when used in more than 200ppm concentration within 72-96h. In addition, the consortium was able to decolorize 52.19% mixed dyes (100ppm) and44.55% Acid blue 113 when used at a concentration as high as 1100ppm within 96h. Optimization of various nutritional and environmental parameters revealed that glucose and yeast extract were the preferred carbon and nitrogen source, respectively,and analysis of treated dye products using high-performance liquid chromatography (HPLC), Fourier-transform infrared spectroscopy (FTIR), and gas chromatography-mass spectrometry (GC-MS) confirmed the breakdown of dye. In all, we present a bacterial consortium with a good ability of dye decolorization that can be used for degrading a wide variety of azo dyes.

Rewiring of Gene Expression in Pseudomonas aeruginosa During Diauxic Growth Reveals an Indirect Regulation of the MexGHI-OpmD Efflux Pump by Hfq.

In Pseudomonas aeruginosa, the RNA chaperone Hfq and the catabolite repression protein Crc act in concert to regulate numerous genes during carbon catabolite repression (CCR). After alleviation of CCR, the RNA CrcZ sequesters Hfq/Crc, which leads to a rewiring of gene expression to ensure the consumption of less preferred carbon and nitrogen sources. Here, we performed a multiomics approach by assessing the transcriptome, translatome, and proteome in parallel in P. aeruginosa strain O1 during and after relief of CCR. As Hfq function is impeded by the RNA CrcZ upon relief of CCR, and Hfq is known to impact antibiotic susceptibility in P. aeruginosa, emphasis was laid on links between CCR and antibiotic susceptibility. To this end, we show that the mexGHI-opmD operon encoding an efflux pump for the antibiotic norfloxacin and the virulence factor 5-Methyl-phenazine is upregulated after alleviation of CCR, resulting in a decreased susceptibility to the antibiotic norfloxacin. A model for indirect regulation of the mexGHI-opmD operon by Hfq is presented.

Sequential uptake of aldoses over fructose and enhanced phosphate solubilization in Rhizobium sp. RM.

Rhizobium sp. RM solubilized tri-calcium phosphate (TCP: 324-463µgml-1) and rock phosphate (RP: 36-46.58µgml-1) in the presence of common rhizospheric sugars-glucose, arabinose, xylose and their combinations. Fructose, though did not support RP solubilization individually, surprisingly solubilized significantly higher phosphate when combined with aldoses. The highest TCP (644µgml-1) and RP (75µgml-1) solubilization was achieved in fructose + glucose combination. Presence of gluconate, malate and oxalate in culture supernatant indicated functioning of periplasmic glucose oxidation, the non-phosphorylative arabinose dehydrogenase pathway and the tricarboxylate (TCA) cycle, respectively. Aldoses, when present together, were co-utilized (monoauxic growth) however, when added with fructose, prevented the uptake of fructose yielding a typical diauxic growth. This presented an unusual sequential utilization of aldoses over a ketose (fructose) in strain RM. The prevention of fructose uptake by aldoses was investigated through real-time expression of key genes coding fructose transport proteins and initial enzymes of sugar metabolism. Fructose was actively transported via fructose-specific ABC transporters as suggested by upregulation of frcB and frcC only in fructose and fructose growth phases of fructose + aldose combinations. The probable route of initial fructose metabolism involved either fructokinase and/or xylose isomerase, as confirmed by enzyme activities. The upregulation of hfq and hprK genes only in aldose phase of fructose + aldose combinations suggested their possible involvement in governing the preferential utilization. The novel aspects of this study are enhanced organic acid mediated P solubilization in fructose + aldose combinations and a rare hierarchy of aldoses over fructose which is possibly regulated at the level of fructose transport and fructokinase. KEY POINTS: • Sugars when provided in different dual combinations, supported enhanced P solubilization from complex phosphate sources like TCP and RP in Rhizobium sp. RM. • Transcriptional status of genes in cells of RM when grown in different individual sugars and their combinations suggested that fructose might be a less preferred carbon source and hence was utilized after aldoses with the possible regulation by Hfq and HPrK. • First study to present a unique phenomenon of sequential utilization of aldoses (glucose, arabinose and xylose) over fructose in a concentration-independent manner in Rhizobium sp. RM. and to present the effect of dual combinations of sugars on organic acid mediated P solubilization trait of rhizobia.

Estimating Preferred Alkane Carbon Numbers of Nonionic Surfactants in Normalized Hydrophilic-Lipophilic Deviation Theory from Dissipative Particle Dynamics Modeling.

The preferred alkane carbon number (PACN) in the normalized hydrophilic-lipophilic deviation (HLDN) theory is a numerical parameter and a transferable scale to characterize the amphiphilicity of surfactants, which is usually measured experimentally using the fish diagram or phase inversion temperature (PIT) methods, and the experimental measurement can only be applied to existing surfactants. Here, for the first time, we propose a procedure to estimate the PACN of CiEj nonionic surfactants directly from dissipative particle dynamics (DPD) simulation. The procedure leverages the method of moment concept to quantitatively evaluate the bending tendency of nonionic surfactant monolayers by calculating the torque density. Seven nonionic surfactants, CiEj (C6E2, C6E3, C8E3, C8E4, C10E4, C12E4, and C12E5), with known PACNs are modeled. Two surfactants, C10E4 and C6E2, were first selected to train and test the interaction parameters, and the relationship between interaction parameters and torque density was mapped for the C10E4-octane-water system using the artificial neural network (ANN) fitting approach to derive the interaction parameters giving zero torque density, then the interaction parameters were tested in the C6E2-dodecane-water system to get the final tuned interaction parameters for PACN estimation. With this procedure, we reproduce the PACN values and their trend of seven nonionic surfactants with reasonable accuracy, which opens the door for quantitative comparison of surfactant amphiphilicity and surfactant classification in silico using the PACN as a transferrable scale.