Compatible Solutes Research Articles

Compatibility and Physical Properties of Dexamethasone-Ondansetron Intravenous Admixture.

Purpose: This work aimed at evaluating the impact of different concentrations and final volumes on the compatibility and physical properties of dexamethasone-ondansetron intravenous (IV) admixture. Methods: The IV admixture of dexamethasone-ondansetron was prepared at different concentrations using normal saline solution as solvent. The final volume of the IV admixture was prepared at 50 and 100 ml. Turbidity was measured as an indicator of physical compatibility of the IV admixture of dexamethasone-ondansetron using UV-visible spectrophotometer and the pH of the IV admixture was measured on day 0, 7,14, and, 21 as an index of chemical stability. Also, the particle size and potential molecular interactions of the admixtures were determined using particle size analyzer and Fourier Transform infrared spectrometry analysis, respectively. Additionally, the effect of preservatives on the IV admixture was also evaluated. Results: Precipitation was observed for mixtures with amounts of dexamethasone and ondansetron exceeding 8 and 16 mg, respectively, in a final volume of 50 ml. For all mixtures with final volume of 100 ml, clear solutions void of any precipitates were observed. The pH of the solution had no effect on the precipitation of the dexamethasone-ondansetron during storage up to 21 days. Analyses of the precipitate formed revealed the presence of molecular interactions between dexamethasone and ondansetron. The benzyl alcohol used as a preservative affected the compatibility of the IV admixture compatibility. Conclusion: Thus, for the preparation of clear, physically compatible normal saline solutions of dexamethasone-ondansetron IV admixture, the maximum amounts of the respective drugs should not exceed 8 and 16 mg in total volume of 50 ml or 20 and 16 mg in a final volume of 100 ml.

Nanoparticles as potential hallmarks of drought stress tolerance in plants.

Plants are inevitably exposed to drought stress limiting their growth and causing yield loss, thus inciting food crises across the world. Nanoparticles (NPs) are regarded as effective and promising tools for modulation of crop yield to overcome current and future constraints in sustainable agricultural production by upgrading the plant tolerance mechanism under abiotic stress conditions, including drought. NPs exhibit alleviating effects against drought stress via induction of physiological and biochemical readjustments accompanied by modulation of gene expression involved in drought response/tolerance. NPs ameliorate drought-induced reduction in carbon assimilation via increasing the photosynthetic activity. The improved root growth, upregulation of aquaporins, modification of intracellular water metabolism, accumulation of compatible solutes and ion homeostasis are the major mechanisms used by NPs to mitigate the osmotic stress caused by water deficit. NPs reduce water loss from leaves through stomatal closure due to fostered abscisic acid (ABA) accumulation and ameliorate oxidative stress damage by reducing reactive oxygen species and activating the antioxidant defense system. This review provides an evolutionary foundation regarding drought stress in plant life and summarizes the interactions between NPs and plants under drought. The subsequent impact of NPs on plant development and productivity and recent nanobiotechnological approaches to improve drought stress resilience are presented. On the whole, this review highlights the significance of NPs in dealing with the global problem of water scarcity faced by farmers.

Oceanographic setting influences the prokaryotic community and metabolome in deep-sea sponges

Marine sponges (phylum Porifera) are leading organisms for the discovery of bioactive compounds from nature. Their often rich and species-specific microbiota is hypothesised to be producing many of these compounds. Yet, environmental influences on the sponge-associated microbiota and bioactive compound production remain elusive. Here, we investigated the changes of microbiota and metabolomes in sponges along a depth range of 1232 m. Using 16S rRNA gene amplicon sequencing and untargeted metabolomics, we assessed prokaryotic and chemical diversities in three deep-sea sponge species: Geodia barretti, Stryphnus fortis, and Weberella bursa. Both prokaryotic communities and metabolome varied significantly with depth, which we hypothesized to be the effect of different water masses. Up to 35.5% of microbial ASVs (amplicon sequence variants) showed significant changes with depth while phylum-level composition of host microbiome remained unchanged. The metabolome varied with depth, with relative quantities of known bioactive compounds increasing or decreasing strongly. Other metabolites varying with depth were compatible solutes regulating osmolarity of the cells. Correlations between prokaryotic community and the bioactive compounds in G. barretti suggested members of Acidobacteria, Proteobacteria, Chloroflexi, or an unclassified prokaryote as potential producers.

A Salt-Tolerant Streptomyces paradoxus D2-8 from Rhizosphere Soil of Phragmites communis Augments Soybean Tolerance to Soda Saline-Alkali Stress.

Soil salinity and alkalization limit plant growth and agricultural productivity worldwide. The application of salt-tolerant plant growth-promoting rhizobacteria (PGPR) effectively improved plant tolerance to saline-alkali stress. To obtain the beneficial actinomyces resources with salt tolerance, thirteen isolates were isolated from rhizosphere saline and alkaline soil of Phragmites communis. Among these isolates, D2-8 was moderately halophilic to NaCl and showed 120 mmol soda saline-alkali solution tolerance. Moreover, the plant growth-promoting test demonstrated that D2-8 produced siderophore, IAA, 1-aminocyclopropane-1-carboxylate deaminase (ACCD), and organic acids. D2-8 showed 99.4% homology with the type strain Streptomyces paradoxus NBRC 14887T and shared the same branch, and, therefore, it was designated S. paradoxus D2-8. Its genome was sequenced to gain insight into the mechanism of growth-promoting and saline-alkali tolerance of D2-8. IAA and siderophore biosynthesis pathway, genes encoding ACC deaminase, together with six antibiotics biosynthesis gene clusters with antifungal or antibacterial activity, were identified. The compatible solute ectoine biosynthesis gene cluster, production, and uptake of choline and glycine betaine cluster in the D2-8 genome may contribute to the saline-alkali tolerance of the strain. Furthermore, D2-8 significantly promoted the seedling growth even under soda saline-alkali stress, and seed coating with D2-8 isolate increased by 5.88% of the soybean yield in the field. These results imply its significant potential to improve soybean soda saline-alkali tolerance and promote crop health in alkaline soil.

The Effects of Exogenous Salicylic Acid on Endogenous Phytohormone Status in Hordeum vulgare L. under Salt Stress.

Acclimation to salt stress in plants is regulated by complex signaling pathways involving endogenous phytohormones. The signaling role of salicylic acid (SA) in regulating crosstalk between endogenous plant growth regulators’ levels was investigated in barley (Hordeum vulgare L. ‘Ince’; 2n = 14) leaves and roots under salt stress. Salinity (150 and 300 mM NaCl) markedly reduced leaf relative water content (RWC), growth parameters, and leaf water potential (LWP), but increased proline levels in both vegetative organs. Exogenous SA treatment did not significantly affect salt-induced negative effects on RWC, LWP, and growth parameters but increased the leaf proline content of plants under 150 mM salt stress by 23.1%, suggesting that SA enhances the accumulation of proline, which acts as a compatible solute that helps preserve the leaf’s water status under salt stress. Changes in endogenous phytohormone levels were also investigated to identify agents that may be involved in responses to increased salinity and exogenous SA. Salt stress strongly affected endogenous cytokinin (CK) levels in both vegetative organs, increasing the concentrations of CK free bases, ribosides, and nucleotides. Indole-3-acetic acid (IAA, auxin) levels were largely unaffected by salinity alone, especially in barley leaves, but SA strongly increased IAA levels in leaves at high salt concentration and suppressed salinity-induced reductions in IAA levels in roots. Salt stress also significantly increased abscisic acid (ABA) and ethylene levels; the magnitude of this increase was reduced by treatment with exogenous SA. Both salinity and SA treatment reduced jasmonic acid (JA) levels at 300 mM NaCl but had little effect at 150 mM NaCl, especially in leaves. These results indicate that under high salinity, SA has antagonistic effects on levels of ABA, JA, ethylene, and most CKs, as well as basic morphological and physiological parameters, but has a synergistic effect on IAA, which was well exhibited by principal component analysis (PCA).

Comprehensive Genome Analysis of Halolamina pelagica CDK2: Insights into Abiotic Stress Tolerance Genes

Halophilic archaeon Halolamina pelagica CDK2, showcasing plant growth-promoting properties and endurance towards harsh environmental conditions (high salinity, heavy metals, high temperature and UV radiation) was sequenced earlier. Pan-genome of Halolamina genus was created and investigated for strain-specific genes of CDK2, which might confer it with features helping it to withstand high abiotic stress. Pathways and subsystems in CDK2 were compared with other Halolamina strain CGHMS and analysed using KEGG and RAST. A genome-scale metabolic model was reconstructed from the genome of H. pelagica CDK2. Results implicated strain-specific genes like thermostable carboxypeptidase and DNA repair protein MutS which might protect the proteins and DNA from high temperature and UV denaturation respectively. A bifunctional trehalose synthase gene responsible for trehalose biosynthesis was also annotated specifying the need for low salt compatible solute strategy, the probable reason behind the ability of this haloarchaea to survive in a wide range of salt concentrations. A modified shikimate and mevalonate pathways were also identified in CDK2, along with many ABC transporters for metal uptakes like zinc and cobalt through pathway analysis. Probable employment of one multifunctional ABC transporter in place of two for similar metals (Nickel/cobalt and molybdenum/tungsten) might be employed as a strategy for energy conservation. The findings of the present study could be utilized for future research relating metabolic model for flux balance analysis and the genetic repertoire imparting resistance to harsh conditions can be transferred to crops for improving their tolerance to abiotic stresses.

Cellular adaptation of Clostridioides difficile to high salinity encompasses a compatible solute-responsive change in cell morphology.

Infections by the pathogenic gut bacterium Clostridioides difficile cause severe diarrhoeas up to a toxic megacolon and are currently among the major causes of lethal bacterial infections. Successful bacterial propagation in the gut is strongly associated with the adaptation to changing nutrition-caused environmental conditions; e.g. environmental salt stresses. Concentrations of 350 mM NaCl, the prevailing salinity in the colon, led to significantly reduced growth of C. difficile. Metabolomics of salt-stressed bacteria revealed a major reduction of the central energy generation pathways, including the Stickland-fermentation reactions. No obvious synthesis of compatible solutes was observed up to 24 h of growth. The ensuing limited tolerance to high salinity and absence of compatible solute synthesis might result from an evolutionary adaptation to the exclusive life of C. difficile in the mammalian gut. Addition of the compatible solutes carnitine, glycine-betaine, γ-butyrobetaine, crotonobetaine, homobetaine, proline-betaine and dimethylsulfoniopropionate restored growth (choline and proline failed) under conditions of high salinity. A bioinformatically identified OpuF-type ABC-transporter imported most of the used compatible solutes. A long-term adaptation after 48 h included a shift of the Stickland fermentation-based energy metabolism from the utilization to the accumulation of l-proline and resulted in restored growth. Surprisingly, salt stress resulted in the formation of coccoid C. difficile cells instead of the typical rod-shaped cells, a process reverted by the addition of several compatible solutes. Hence, compatible solute import via OpuF is the major immediate adaptation strategy of C. difficile to high salinity-incurred cellular stress.

Impact of Two Strains of Rhizobium leguminosarum on the Adaptation to Terminal Water Deficit of Two Cultivars Vicia faba.

Drought stress has become one of the most uncontrolled and unpredictable constraints on crop production. The purpose of this study was to evaluate the impacts of two different Rhizobium leguminosarum strains on terminal drought tolerance induction in two faba bean genotypes cultivated in Algeria, Aquadulce and Maltais. To this end, we measured physiological parameters—osmoprotectants accumulation, oxidative stress markers and enzyme activities—to assess the effect of R. leguminosarum inoculation on V. faba under terminal water deficiency conditions in greenhouse trials. Upregulation of anti-oxidative mechanisms and production of compatible solutes were found differentially activated according to Rhizobium strain. Drought stress resilience of the Maltais variety was improved using the local Rhizobium strain OL13 compared to the common strain 3841. Symbiosis with OL13 strain leads in particular to a much better production of proline and soluble sugar in nodules but also in roots and leaves of Maltais plant. Even if additional work is still necessary to decipher the mechanism by which a Rhizobium strain can affect the accumulation of osmoprotectants or cellular redox status in all the plants, inoculation with selected Rhizobium could be a promising strategy for improving water stress management in the forthcoming era of climate change.

Integrative genome and metabolome analysis reveal the potential mechanism of osmotic stress tolerance in Bifidobacterium bifidum

Osmotic stress affects the viability of probiotics, especially Bifidobacteria, which are less resistant to environmental stress. In this study, an adaptive evolution technique was applied to Bifidobacterium bifidum to select tolerant strains against high osmotic stress. B. bifidum CCFM16 was subjected to a gradually increasing penetration environment for 1000 generations. The generation time of the hyperosmotic tolerance mutant CCFM16m is 1/3 of its ancestor at an osmotic stress of 1300 mOsm/kg. Whole-genome comparison, candidate gene expression, and 3D protein structure simulation were performed and the results indicated that critical mutations were related to glutamate synthesis and transport. Untargeted metabolome analysis showed that compared to parent strain, the difference was mainly reflected in the increased accumulation of amino acids, especially proline, while the accumulation of central carbon decreased. Increase in the key enzyme F6PPK in the “bifidus” pathway indicated that cells transfer more energy from basal metabolism to resist osmotic stress. Since there was no proline transporter protein, excessive accumulation of proline showed that under long-term high osmotic stress, B. bifidum CCFM16 evolved a protective mechanism to synthesize proline from glutamic acid, an osmotic protection system with proline as the main compatible solute was formed.

Microwave-induced self-ignition: An efficient approach for high purity graphite digestion and multitechnique halogen determination

In this work, high purity graphite, a high chemically stable material, was effectively digested using a single method allowing compatible solutions for the further multitechnique determination of halogens by: ion chromatography (F and Cl), inductively coupled plasma mass spectrometry (Cl, Br and I) and by ion selective electrode (only for F). The recent system using microwave-induced self-ignition (MISI) is based on the strong interaction between microwave radiation and graphite in a closed system pressurized with oxygen (Maxwell-Wagner effect). Carbon-based materials present intense and specific interfacial polarization when exposed to microwave electromagnetic field resulting in a fast heating rate. This effect associated to a pressurized oxygen system, provides a quick self-ignition of carbon-based materials and consequent combustion/digestion of organic matrices. Under optimized conditions, sample masses up to 600 mg were fully digested in a quartz vessel under 20 bar of oxygen pressure and using just a diluted solution (100 mmol L−1 NH4OH) for the quantitative absorption of all the analytes. MISI method was validated, and the accuracy (better than 94%) was evaluated by comparison of results obtained by pyrohydrolysis for two coal certified reference materials as well as with subsequent analytes determination by the three techniques: IC, ICP-MS and ISE. It is important to point out that no filter paper disks, electrical connections or other ignition aids are required as in the case of previous or classical combustion methods. Moreover, just a diluted absorbing solution was used resulting in negligible blanks and relatively low limits of detection. The digestion efficiency was higher than 99%, making the proposed method a suitable and powerful alternative for the quasi complete digestion of graphite and determination of halogens virtually free of interferences.

PGPR-Mediated Salt Tolerance in Maize by Modulating Plant Physiology, Antioxidant Defense, Compatible Solutes Accumulation and Bio-Surfactant Producing Genes.

Salinity stress is a barrier to crop production, quality yield, and sustainable agriculture. The current study investigated the plant growth promotion, biochemical and molecular characterization of bacterial strain Enterobacter cloacae PM23 under salinity stress (i.e., 0, 300, 600, and 900 mM). E. cloacae PM23 showed tolerance of up to 3 M NaCl when subjected to salinity stress. Antibiotic-resistant Iturin C (ItuC) and bio-surfactant-producing genes (sfp and srfAA) were amplified in E. cloacae PM23, indicating its multi-stress resistance potential under biotic and abiotic stresses. Moreover, the upregulation of stress-related genes (APX and SOD) helped to mitigate salinity stress and improved plant growth. Inoculation of E. cloacae PM23 enhanced plant growth, biomass, and photosynthetic pigments under salinity stress. Bacterial strain E. cloacae PM23 showed distinctive salinity tolerance and plant growth-promoting traits such as indole-3-acetic acid (IAA), siderophore, ACC deaminase, and exopolysaccharides production under salinity stress. To alleviate salinity stress, E. cloacae PM23 inoculation enhanced radical scavenging capacity, relative water content, soluble sugars, proteins, total phenolic, and flavonoid content in maize compared to uninoculated (control) plants. Moreover, elevated levels of antioxidant enzymes and osmoprotectants (Free amino acids, glycine betaine, and proline) were noticed in E. cloacae PM23 inoculated plants compared to control plants. The inoculation of E. cloacae PM23 significantly reduced oxidative stress markers under salinity stress. These findings suggest that multi-stress tolerant E. cloacae PM23 could enhance plant growth by mitigating salt stress and provide a baseline and ecofriendly approach to address salinity stress for sustainable agriculture.

Fluoride tolerance in rice is negatively regulated by the 'stress-phytohormone' abscisic acid (ABA), but promoted by ABA-antagonist growth regulators, melatonin, and gibberellic acid.

The present manuscript aimed at investigating whether abscisic acid (ABA) promotes fluoride tolerance, similar to inciting salt adaptation in rice. Seeds of three salt-tolerant rice genotypes were maintained at 32°C under 16/8h light/dark photoperiodic cycle with 700μmol photons m-2s-1 intensity and 50% relative humidity in a plant growth chamber for 20days. Suppressed ABA biosynthesis, and downregulated expression of ABA-inducible genes like Rab16A, Osem, and TRAB1 triggered NaCl-induced growth inhibition and physiological injuries like chlorophyll degradation, electrolyte leakage, formation of H2O2, malondialdehyde, and methylglyoxal in Matla. Reduced ABA accumulation increased the levels of melatonin and gibberellic acid in NaF (50mgL-1)-stressed Nonabokra and Matla, which altogether promoted fluoride tolerance. Higher ABA content in NaF-stressed Jarava stimulated fluoride uptake via chloride channels, thus exhibiting severe fluoride susceptibility, in spite of higher production of ABA-associated osmolytes like proline, glycine-betaine and polyamines via the concerted action of genes like PDH, ADC, ODC, SAMDC, SPDS, SPMS, DAO, and PAO. Increased accumulation of compatible solutes in presence of high endogenous ABA promoted salt tolerance in Jarava; the same was insufficient to ameliorate fluoride-induced injuries in this cultivar. Treatment with ABA biosynthetic inhibitor, Na2WO4 promoted fluoride tolerance in Jarava, whereas further supplementation with exogenous ABA resulted in reversion back to fluoride-susceptible phenotype. Our work clearly established that ABA cannot always be considered as a 'universal' stress hormone as known in literature, since it acts as a negative regulator of fluoride tolerance which is more tightly regulated in rice via melatonin- and gibberellic acid-dependent pathways in ABA-independent manner.

Insights into Ionic liquids-resistance mechanism and lignocellulose-degradation model of Aspergillus terreus in 1-ethyl-3-methylimidazolium acetate

Ionic liquids (ILs)-assisted microbes and enzymes are potential approaches for converting lignocellulosic biomass into fermentable sugars in a biorefinery. In the present study, an ILs-tolerant strain Aspergillus terreus NEAU-7 with ILs-stable cellulase production ability was isolated and identified. A comparative transcriptome analysis revealed that A. terreus NEAU-7 could promote ion transport and acquire metabolic energy to reduce the adverse effect of ILs. The results further confirmed that the strain could increase the accumulation of compatible solutes to offset the osmotic pressure of ILs and reduce the cell membrane permeability to prevent ILs. A secretome analysis of Carbohydrate-Active Enzymes (CAZymes) showed no significant reduction in the Glycoside Hydrolases (GH) families of cellulolytic enzymes, while a significant reduction was observed for Carbohydrate Esterases (CE) families and Glycoside Hydrolase 43 (encoding α-L-arabinofuranosidases and β-xylosidases), indicating that the presence of ILs could decrease the expression of hemicellulolytic enzymes. Herein, a model for lignocellulose degradation of A. terreus NEAU-7 was interpreted. Overall, this study will provide insights into further exploration of the ILs-resistance mechanism of lignocellulolytic fungi and employment of ILs-assisted fungi in the degradation of lignocellulosic biomass into value-added products.

Direct Electromembrane Extraction‐Based Mass Spectrometry: A Tool for Studying Drug Metabolism Properties of Liver Organoids

AbstractThis work introduces a strategy for organoid analysis ‐ direct Electromembrane Extraction based Mass Spectrometry (dEME‐MS) – for coupling liver organoids with mass spectrometry (MS). dEME‐MS comprises electrophoresis of selected small molecules from a culture chamber across an oil membrane, and to a MS compatible solution. This enables clean micro‐extraction of drugs and their metabolites as produced in the liver organoids to capillary liquid chromatography‐mass spectrometry. Applying dEME‐MS, proof‐of‐concept of directly measuring methadone metabolism is demonstrated on adult liver organoids. With 50 liver organoids and 1 μM methadone, methadone metabolism was monitored from 0 to 24 hours (11 time points). All analytes had <0.4 % variance in retention times with >100 measurements. dEME‐MS is capable of automated and selective monitoring of drug metabolism in liver organoids, and could serve as a valuable tool for automated drug discovery efforts.

Methylphosphonate Degradation and Salt-Tolerance Genes of Two Novel Halophilic Marivita Metagenome-Assembled Genomes from Unrestored Solar Salterns.

Aerobic bacteria that degrade methylphosphonates and produce methane as a byproduct have emerged as key players in marine carbon and phosphorus cycles. Here, we present two new draft genome sequences of the genus Marivita that were assembled from metagenomes from hypersaline former industrial salterns and compare them to five other Marivita reference genomes. Phylogenetic analyses suggest that both of these metagenome-assembled genomes (MAGs) represent new species in the genus. Average nucleotide identities to the closest taxon were <85%. The MAGs were assembled with SPAdes, binned with MetaBAT, and curated with scaffold extension and reassembly. Both genomes contained the phnCDEGHIJLMP suite of genes encoding the full C-P lyase pathway of methylphosphonate degradation and were significantly more abundant in two former industrial salterns than in nearby reference and restored wetlands, which have lower salinity levels and lower methane emissions than the salterns. These organisms contain a variety of compatible solute biosynthesis and transporter genes to cope with high salinity levels but harbor only slightly acidic proteomes (mean isoelectric point of 6.48).

Enhanced production of ectoine from methane using metabolically engineered Methylomicrobium alcaliphilum 20Z

BackgroundEctoine (1,3,4,5-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid) is an attractive compatible solute because of its wide industrial applications. Previous studies on the microbial production of ectoine have focused on sugar fermentation. Alternatively, methane can be used as an inexpensive and abundant resource for ectoine production by using the halophilic methanotroph, Methylomicrobium alcaliphilum 20Z. However, there are some limitations, including the low production of ectoine from methane and the limited tools for the genetic manipulation of methanotrophs to facilitate their use as industrial strains.ResultsWe constructed M. alcaliphilum 20ZDP with a high conjugation efficiency and stability of the episomal plasmid by the removal of its native plasmid. To improve the ectoine production in M. alcaliphilum 20Z from methane, the ectD (encoding ectoine hydroxylase) and ectR (transcription repressor of the ectABC-ask operon) were deleted to reduce the formation of by-products (such as hydroxyectoine) and induce ectoine production. When the double mutant was batch cultured with methane, ectoine production was enhanced 1.6-fold compared to that obtained with M. alcaliphilum 20ZDP (45.58 mg/L vs. 27.26 mg/L) without growth inhibition. Notably, a maximum titer of 142.32 mg/L was reached by the use of an optimized medium for ectoine production containing 6% NaCl and 0.05 μM of tungsten without hydroxyectoine production. This result demonstrates the highest ectoine production from methane to date.ConclusionsEctoine production was significantly enhanced by the disruption of the ectD and ectR genes in M. alcaliphilum 20Z under optimized conditions favoring ectoine accumulation. We demonstrated effective genetic engineering in a methanotrophic bacterium, with enhanced production of ectoine from methane as the sole carbon source. This study suggests a potentially transformational path to commercial sugar-based ectoine production.Graphical

Heterologous Production of Glycine Betaine Using Synechocystis sp. PCC 6803-Based Chassis Lacking Native Compatible Solutes.

Among compatible solutes, glycine betaine has various applications in the fields of nutrition, pharmaceuticals, and cosmetics. Currently, this compound can be extracted from sugar beet plants or obtained by chemical synthesis, resulting in low yields or high carbon footprint, respectively. Hence, in this work we aimed at exploring the production of glycine betaine using the unicellular cyanobacterium Synechocystis sp. PCC 6803 as a photoautotrophic chassis. Synechocystis mutants lacking the native compatible solutes sucrose or/and glucosylglycerol—∆sps, ∆ggpS, and ∆sps∆ggpS—were generated and characterized. Under salt stress conditions, the growth was impaired and accumulation of glycogen decreased by ∼50% whereas the production of compatible solutes and extracellular polymeric substances (capsular and released ones) increased with salinity. These mutants were used as chassis for the implementation of a synthetic device based on the metabolic pathway described for the halophilic cyanobacterium Aphanothece halophytica for the production of the compatible solute glycine betaine. Transcription of ORFs comprising the device was shown to be stable and insulated from Synechocystis’ native regulatory network. Production of glycine betaine was achieved in all chassis tested, and was shown to increase with salinity. The introduction of the glycine betaine synthetic device into the ∆ggpS background improved its growth and enabled survival under 5% NaCl, which was not observed in the absence of the device. The maximum glycine betaine production [64.29 µmol/gDW (1.89 µmol/mg protein)] was reached in the ∆ggpS chassis grown under 3% NaCl. Taking into consideration this production under seawater-like salinity, and the identification of main key players involved in the carbon fluxes, this work paves the way for a feasible production of this, or other compatible solutes, using optimized Synechocystis chassis in a pilot-scale.

Time-space-direction Extension TRIZ Innovation Model for Product Innovation

In order to solve the problem that college students are prone to thinking set and direction deviation in the process of innovation practice. TRIZ (Latin abbreviation of "Teoriya Resheniya Izoblatelskikh Zadatch", which means theory of the solution of innovative problems) is extended and matched with the meta conditional features or quantities of the three dimensions of TSD (time, space and direction) of the problem. In this paper, TRIZ-TSD extension problem solving model is proposed to find compatible solutions. TRIZ-TSD extension problem solving model expands the available resources of the original TRIZ analysis tools, strengthens the interaction between the analysis tools, and makes it more suitable for beginners to use in practical innovation. Taking the university student innovation award-winning project "the intelligent wall planting system" as an example, the basic process and practical effect of TRIZ-TSD fusion innovation model in solving specific problems are verified. The basic principle and thinking mode of this method is not only limited to the practice of College Students’ innovation projects, but also has certain reference value for solving problems in other fields.

Identification and characterization of a novel GNAT superfamily Nα -acetyltransferase from Salinicoccus halodurans H3B36.

SummaryNα‐acetyl‐α‐lysine was found as a new type of compatible solutes that acted as an organic cytoprotectant in the strain of Salinicoccus halodurans H3B36. A novel lysine Nα‐acetyltransferase gene (shkat), encoding an enzyme that catalysed the acetylation of lysine exclusively at α position, was identified from this moderate halophilic strain and expressed in Escherichia coli. Sequence analysis indicated ShKAT contained a highly conserved pyrophosphate‐binding loop (Arg‐Gly‐Asn‐Gly‐Asn‐Gly), which was a signature of the GNAT superfamily. ShKAT exclusively recognized free amino acids as substrate, including lysine and other basic amino acids. The enzyme showed a wide range of optimal pH value and was tolerant to high‐alkali and high‐salinity conditions. As a new member of the GNAT superfamily, the ShKAT was the first enzyme recognized free lysine as substrate. We believe this work gives an expanded perspective of the GNAT superfamily, and reveals great potential of the shkat gene to be applied in genetic engineering for resisting extreme conditions.

All-fiber-transmission photometry for simultaneous optogenetic stimulation and multi-color neuronal activity recording

Manipulating and real-time monitoring of neuronal activities with cell-type specificity and precise spatiotemporal resolution during animal behavior are fundamental technologies for exploring the functional connectivity, information transmission, and physiological functions of neural circuits<italic>in vivo</italic>. However, current techniques for optogenetic stimulation and neuronal activity recording mostly operate independently. Here, we report an all-fiber-transmission photometry system for simultaneous optogenetic manipulation and multi-color recording of neuronal activities and the neurotransmitter release in a freely moving animal. We have designed and manufactured a wavelength-independent multi-branch fiber bundle to enable simultaneous optogenetic manipulation and multi-color recording at different wavelengths. Further, we combine a laser of narrow linewidth with the lock-in amplification method to suppress the optogenetic stimulation-induced artifacts and channel crosstalk. We show that the collection efficiency of our system outperforms a traditional epi-fluorescence system. Further, we demonstrate successful recording of dynamic dopamine (DA) responses to unexpected rewards in the nucleus accumbens (NAc) in a freely moving mouse. We also show simultaneous dual-color recording of neuronal Ca²⁺ signals and DA dynamics in the NAc upon delivering an unexpected reward and the simultaneous optogenetic activating at dopaminergic terminals in the same location. Thus, our multi-function fiber photometry system provides a compatible, efficient, and flexible solution for neuroscientists to study neural circuits and neurological diseases.