ABSTRACT
Cadaverine is a fundamental C5 building block in the production of polyamides. Due to the limited regeneration efficiency of intracellular pyridoxal 5'-phosphate (PLP), the current fermentation-based production of cadaverine exhibits low efficiency. In this study, we developed an Escherichia coli strain L01 by introducing lysine decarboxylase (lysine decarboxylase, LDC, a key enzyme in the synthesis of cadaverine) into a lysine-producing strain E. coli LY-4, achieving a cadaverine tier of 1.07 g/L in shake flask fermentation. Subsequently, a dual metabolic pathway enhancement strategy was proposed to synergistically strengthen both endogenous and exogenous PLP synthesis modules, thereby improving intracellular PLP synthesis. The optimized strain L11 achieved a cadaverine titer of 9.23 g/L in shake flask fermentation. Finally, the fermentation process for cadaverine production by strain L11 was optimized in a 5 L fermenter. After 48 h of fed-batch fermentation, the engineered strain L11 achieved the cadaverine titer, yield, and productivity of 54.43 g/L, 0.22 g/g, and 1.13 g/(L·h), respectively. This study provides a theoretical and technical foundation for establishing microbial cell factories for bioamine production.
Subject(s)
Cadaverine , Carboxy-Lyases , Escherichia coli , Fermentation , Metabolic Engineering , Pyridoxal Phosphate , Cadaverine/biosynthesis , Cadaverine/metabolism , Metabolic Engineering/methods , Escherichia coli/metabolism , Escherichia coli/genetics , Carboxy-Lyases/genetics , Carboxy-Lyases/metabolism , Pyridoxal Phosphate/metabolismABSTRACT
Cadaverine is an endogenous metabolite produced by the gut microbiome with various activity in physiological and pathological conditions. However, whether cadaverine regulates pain or itch remains unclear. In this study, we first found that cadaverine may bind to histamine 4 receptor (H4R) with higher docking energy score using molecular docking simulations, suggesting cadaverine may act as an endogenous ligand for H4R. We subsequently found intradermal injection of cadaverine into the nape or cheek of mice induces a dose-dependent scratching response in mice, which was suppressed by a selective H4R antagonist JNJ-7777120, transient receptor potential vanilloid 1 (TRPV1) antagonist capsazepine and PLC inhibitor U73122, but not H1R antagonist or TRPA1 antagonist or TRPV4 antagonist. Consistently, cadaverine-induced itch was abolished in Trpv1-/- but not Trpa1-/- mice. Pharmacological analysis indicated that mast cells and opioid receptors were also involved in cadaverine-induced itch in mice. scRNA-Seq data analysis showed that H4R and TRPV1 are mainly co-expressed on NP2, NP3 and PEP1 DRG neurons. Calcium imaging analysis showed that cadaverine perfusion enhanced calcium influx in the dissociated dorsal root ganglion (DRG) neurons, which was suppressed by JNJ-7777120 and capsazepine, as well as in the DRG neurons from Trpv1-/- mice. Patch-clamp recordings found that cadaverine perfusion significantly increased the excitability of small diameter DRG neurons, and JNJ-7777120 abolished this effect, indicating involvement of H4R. Together, these results provide evidences that cadaverine is a novel endogenous pruritogens, which activates H4R/TRPV1 signaling pathways in the primary sensory neurons.
Subject(s)
Cadaverine , Ganglia, Spinal , Mice, Inbred C57BL , Pruritus , TRPV Cation Channels , Animals , Pruritus/metabolism , Pruritus/chemically induced , TRPV Cation Channels/metabolism , Ganglia, Spinal/metabolism , Ganglia, Spinal/drug effects , Male , Cadaverine/analogs & derivatives , Cadaverine/pharmacology , Cadaverine/metabolism , Mice , Mice, Knockout , Humans , Mast Cells/metabolism , Mast Cells/drug effects , TRPA1 Cation Channel/metabolism , Sensory Receptor Cells/metabolism , Sensory Receptor Cells/drug effects , Receptors, G-Protein-Coupled/metabolism , Capsaicin/analogs & derivativesABSTRACT
Lactic acid bacteria (LAB) are frequently used in meat fermentation, and mixed stater cultures are reported to perform better than single ones. Lactiplantibacillus plantarum 3-19 and Pediococcus pentosaceus 18-1 were chosen from 28 sour-meat-origin strains to examine the effects of single and combined inoculation on sour meat quality. Natural fermentation was used as a control to investigate changes in pH, water activity (aw), amino acid nitrogen (AN), texture, microbial diversity, and volatile organic compounds (VOCs) during fermentation. The pH and aw of each inoculation group were significantly decreased, and AN content was significantly increased. The inoculation of P. pentosaceus 18-1 significantly reduced putrescine, cadaverine, and tryptamine content (p < 0.05), while the inoculation of Lpb. plantarum 3-19 significantly reduced cadaverine amounts (p < 0.05). At the fermentation endpoint, the total biogenic amines content in the C group was 992.96 ± 14.07, which was 1.65, 2.57, and 3.07 times higher than that in the Lp, Pe, and M groups, respectively. The mixed inoculation group combined the advantages of both strains and decreased total biogenic amines most significantly. At the end of fermentation, the VOCs in C, Lp, Pe, and M groups were 10.11, 11.56, 12.45, and 13.39 times higher than those at the beginning of fermentation. Inoculation promoted the production of key VOCs (OAV > 2000) such as heptanal, octanal, and (E)-2-nonanal. The mixed inoculation group had the highest variety and content of VOCs and the highest content of the above key VOCs, significantly enhancing its fruity, floral, ester, and other aromas. Sensory evaluation indicated that the M group had the best overall acceptability. Finally, it was suggested that a combination of Lpb. plantarum 3-19 and P. pentosaceus 18-1 is a novel and efficient starter culture for processing sour meat since they lower the amounts of biogenic amines in the meat and promote the production of VOCs.
Subject(s)
Biogenic Amines , Fermentation , Food Microbiology , Pediococcus pentosaceus , Volatile Organic Compounds , Biogenic Amines/metabolism , Volatile Organic Compounds/metabolism , Volatile Organic Compounds/analysis , Pediococcus pentosaceus/metabolism , Lactobacillus plantarum/metabolism , Meat/microbiology , Humans , Hydrogen-Ion Concentration , Cadaverine/metabolismABSTRACT
Periodontitis is an inflammatory condition of supporting structures of teeth leading to attachment and bone loss. Cigarette smoking is the single most important and modifiable risk factor with 5 to 20-fold susceptibility for periodontal diseases. Reverse smoking is a peculiar habit of smoking where the lit end is kept inside the mouth, which is predominant in the northern coastal districts of Andhra Pradesh. Polyamines are biologically active amines involved in tissue regeneration and modulation of inflammation. The study aimed to evaluate polyamines and check their utility as a marker in detection of periodontitis among different groups. Total polyamine levels showed significant increase in reverse smokers with periodontitis when compared to the other groups. Qualitative analysis by thin layer chromatography showed three polyamine bands with varying intensity among the different groups. Mass spectrometric and NMR analyses of the three bands identified them as N1, N8-diacetyl spermidine, N-acetyl cadaverine and lysine. Most significantly elevated levels of lysine was observed in the smoker and reverse smoker periodontitis groups when compared to healthy and non-smoker periodontitis groups. The significantly elevated levels of N-acetyl cadaverine could be responsible for the more destruction of periodontium in the reverse smoker group. Antioxidant potential decreased significantly in different smoker periodontitis groups. The present study suggests that the quantitative analysis of salivary polyamines, lysine and N-acetyl cadaverine can aid as an easy noninvasive diagnostic method for assessing the periodontal status, especially in smokers.
Subject(s)
Biomarkers , Cadaverine , Lysine , Periodontitis , Humans , Periodontitis/metabolism , Periodontitis/diagnosis , Cadaverine/metabolism , Cadaverine/analysis , Biomarkers/metabolism , Biomarkers/analysis , Lysine/analogs & derivatives , Lysine/analysis , Lysine/metabolism , Adult , Male , Smokers , Female , Middle Aged , Smoking , Saliva/chemistry , Saliva/metabolismABSTRACT
Cadaverine is a critical C5 monomer for the production of polyamides. Pyridoxal 5'-phosphate (PLP), as a crucial cofactor for the key enzyme lysine decarboxylase in the cadaverine biosynthesis pathway, has seen a persistent shortage, leading to limitations in cadaverine production. To address this issue, a dual-pathway strategy was implemented, synergistically enhancing both endogenous and heterologous PLP synthesis modules and resulting in improved PLP synthesis. Subsequently, a growth-stage-dependent molecular switch was introduced to balance the precursor competition between PLP synthesis and cell growth. Additionally, a PLP sensor-based negative feedback circuit was constructed by integrating a newly identified PLP-responsive promoter PygjH and an arabinose-regulated system, dynamically regulating the expression of the PLP synthetic genes and preventing excessive intracellular PLP accumulation. The optimal strain, L18, cultivated in the minimal medium AM1, demonstrated cadaverine production with a titer, yield, and productivity of 64.03 g/L, 0.23 g/g glucose, and 1.33 g/L/h, respectively. This represents the highest titer reported to date in engineered Escherichia coli by fed-batch fermentation in a minimal medium.
Subject(s)
Cadaverine , Culture Media , Escherichia coli , Metabolic Engineering , Pyridoxal Phosphate , Cadaverine/metabolism , Cadaverine/biosynthesis , Pyridoxal Phosphate/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Metabolic Engineering/methods , Culture Media/chemistry , Promoter Regions, Genetic , Carboxy-Lyases/genetics , Carboxy-Lyases/metabolismABSTRACT
Among the compounds present in necro-leachate, a liquid released during the process of decomposition of the human body, are the biogenic amines cadaverine and putrescine. Although some studies on necro-leachate have indicated a potential ecotoxicological and public health risk associated with it, the research on this type of contamination is still rather limited. This study presents information about the phytotoxic and cytogenotoxic potential of cadaverine and putrescine, evaluated separately and within a mixture. Phytotoxicity was evaluated through a germination test, the initial growth of seedlings with Lactuca sativa, and cytogenotoxicity through chromosomal aberration and micronucleus tests with Allium cepa. The L. sativa results showed a phytotoxic effect for the evaluated amines, by reducing root (> 90%) and hypocotyl (> 80%) elongation. The co-exposure of cadaverine and putrescine potentiated cytogenotoxic activity by aneugenic action in the meristematic cells of A. cepa. From this result, it is possible to infer the eco-toxicogenic potential of cadaverine and putrescine. This study not only highlights the importance of the phytotoxic and cytogenotoxic effects of these amines but also emphasizes the urgent need for further investigation into contamination originating from cemetery environments. By evaluating the risks associated with necro-leachate, this research is aimed at informing global efforts to protect ecological and public health.
Subject(s)
Biogenic Amines , Cadaverine , Putrescine , Biogenic Amines/toxicity , Lactuca/drug effects , Onions/drug effects , Germination/drug effectsABSTRACT
Highly polar low molecular weight organic molecules are still very challenging to analyze by liquid chromatography. Yet, with the steadily increasing application of metabolomics and similar approaches in chemical analysis, separating polar compounds might be even more important. However, almost all established liquid chromatography techniques (i.e., normal and reversed phase, hydrophilic interaction liquid chromatography (HILIC), ion chromatography) struggle with either carry-over, low sensitivity, or a lack of retention. For improving these shortcomings, electrostatic repulsion hydrophilic interaction chromatography (ERLIC) might be an alternative. By combining a HILIC mobile phase, that is highly organic with a low water content, and an ion exchange column, a distinct layer system develops. When the analyte's charge is of the same direction as the stationary phase, retention and elution are determined by two antagonistic forces: electrostatic repulsion and hydrophilicity. One prominent group of challenging polar analytes are the polyamines cadaverine, putrescine, spermidine, and spermine. Carrying charges from +2 to +4 at physiological pH, these compounds are essential cell constituents and found in all living organisms. However, they are still notoriously challenging to analyze via the established liquid chromatography methods. In the present work, an ERLIC tandem mass spectrometry method has been exemplarily developed, optimized, and validated for the quantitative determination of cadaverine, putrescine, spermidine, and spermine. This method enables symmetrical peak shapes and good separation of analytes with different charges while simultaneously selectively detecting the co-eluting diamines by MS/MS. Furthermore, high linearity (R > 0.998) and sensitivity (LODs ≤ 2 ng/mL) have been proven. Thus, ERLIC may be interesting for both targeted and untargeted analysis approaches of highly charged low molecular weight organic molecules.
Subject(s)
Polyamines , Tandem Mass Spectrometry , Tandem Mass Spectrometry/methods , Static Electricity , Putrescine , Spermidine , Spermine , Cadaverine , Gas Chromatography-Mass Spectrometry , Chromatography, Liquid/methods , Hydrophobic and Hydrophilic InteractionsABSTRACT
The response patterns of microbial functional genes involved in biogeochemical cycles to cadaver decay is a central topic of recent environmental sciences. However, the response mechanisms and pathways of the functional genes associated with the carbon (C) and nitrogen (N) cycling to cadaveric substances such as cadaverine and putrescine remain unclear. This study explored the variation of functional genes associated with C fixation, C degradation and N cycling and their influencing factors under cadaverine, putrescine and mixed treatments. Our results showed only putrescine significantly increased the alpha diversity of C fixation genes, while reducing the alpha diversity of N cycling genes in sediment. For the C cycling, the mixed treatment significantly decreased the total abundance of reductive acetyl-CoA pathway genes (i.e., acsB and acsE) and lig gene linked to lignin degradation in water, while only significantly increasing the hydroxypropionate-hydroxybutylate cycle (i.e., accA) gene abundance in sediment. For the N cycling, mixed treatment significantly decreased the abundance of the nitrification (i.e., amoB), denitrification (i.e., nirS3) genes in water and the assimilation pathway gene (i.e., gdhA) in sediment. Environmental factors (i.e., total carbon and total nitrogen) were all negatively associated with the genes of C and N cycling. Therefore, cadaverine and putrescine exposure may inhibit the pathway in C fixation and N cycling, while promoting C degradation. These findings can offer some new insight for the management of amine pollution caused by animal cadavers.
Subject(s)
Carbon , Putrescine , Humans , Animals , Cadaverine , Water , Rivers/chemistry , Geologic Sediments/chemistry , Nitrogen Cycle , NitrogenABSTRACT
The biosynthesis of cadaverine from lysine is an environmentally promising technology, that could contribute to a more sustainable approach to manufacturing bio-nylon 5X. However, the titer of biosynthesized cadaverine has still not reached a sufficient level for industrial production. A powerful green cell factory was developed to enhance cadaverine production by regulating lipopolysaccharide (LPS) genes and improving membrane permeability. Firstly, 10 LPS mutant strains were constructed and the effect on the growth was investigated. Then, the lysine decarboxylase (CadA) was overexpressed in 10 LPS mutant strains of Escherichia coli MG1655 and the ability to produce cadaverine was compared. Using 20.0 g L-1 of L-lysine hydrochloride (L-lysine-HCl) as the substrate for the biotransformation reaction, Cad02 and Cad06 strains exhibited high production levels of cadaverine, with 8.95 g L-1 and 7.55 g L-1 respectively while the control strain Cad00 only 4.92 g L-1 . Directed evolution of CadA was also used to improve its stability under alkaline conditions. The cadaverine production of the Cad02-M mutant stain increased by 1.86 times at pH 8.0. Finally, the production process was scaled up using recombinant whole cells as catalysts, achieving a high titer of 211 g L-1 cadaverine (96.8%) by fed-batch bioconversion. This study demonstrates the potential role of LPS in enhancing the efficiency of mass transfer between substrate and enzymes in vivo by increasing cell permeability. The results indicate that the argumentation of cell permeability could not only significantly enhance the biotransformation efficiency of cadaverine, but also provide a universally applicable, straightforward, environment-friendly, and cost-effective method for the biosynthesis of other high-value chemicals.
Subject(s)
Escherichia coli , Lipopolysaccharides , Escherichia coli/genetics , Cadaverine/metabolism , Lipopolysaccharides/metabolism , Catalysis , Biotransformation , Lysine/metabolismABSTRACT
The process of biological fermentation is often accompanied by the release of CO2, resulting in low yield and environmental pollution. Refixing CO2 to the product synthesis pathway is an attractive approach to improve the product yield. Cadaverine is an important diamine used for the synthesis of bio-based polyurethane or polyamide. Here, aiming to increase its final production, a RuBisCO-based shunt consisting of the ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and phosphoribulate kinase (PRK) was expressed in cadaverine-producing E. coli. This shunt was calculated capable of increasing the maximum theoretical cadaverine yield based on flux model analysis. When a functional RuBisCO-based shunt was established and optimized in E. coli, the cadaverine production and yield of the final engineered strain reached the highest level, which were 84.1 g/L and 0.37 g/g Glucose, respectively. Thus, the design of in situ CO2 fixation provides a green and efficient industrial production process.
Subject(s)
Escherichia coli , Ribulose-Bisphosphate Carboxylase , Ribulose-Bisphosphate Carboxylase/metabolism , Cadaverine/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Carbon Dioxide/metabolism , FermentationABSTRACT
Malignant tumors are still one of the most deadly diseases that threaten human life and health. However, developing new drugs is challenging due to lengthy trials, funding constraints, and regulatory approval procedures. Consequently, researchers have devoted themselves to transforming some clinically approved old drugs into antitumor drugs with certain active ingredients, which have become an attractive alternative. Disulfiram (DSF), an antialcohol medication, can rapidly metabolize in the physiological environment into diethyldithiocarbamate (DTC) which can readily react with Cu2+ ions in situ to form the highly toxic bis(N,N-diethyldithiocarbamate)-copper(II) (CuET) complex. In this study, DSF is loaded into mesoporous dopamine nanocarriers and surface-chelated with tannin and Cu2+ to construct M-MDTC nanoprodrugs under the camouflage of K7 tumor cell membranes. After intravenous injection, M-MDTC nanoprodrugs successfully reach the tumor sites with the help of mediated cell membranes. Under slightly acidic pH and photothermal stimulation conditions, DSF and Cu2+ are simultaneously released, forming a highly toxic CuET to kill tumor cells in situ. The generated CuET can also induce immunogenic cell death of tumor cells, increase the proportion of CD86+ CD80+ cells, and promote dendritic cell maturation. In vitro and in vivo studies of M-MDTC nanoprodrugs have shown excellent tumor-cell-killing ability and solid tumor suppression. This approach enables in situ amplification of chemotherapy in the tumor microenvironment, achieving an effective antitumor treatment.
Subject(s)
Cadaverine/analogs & derivatives , Copper , Neoplasms , Humans , Cell Line, Tumor , Copper/pharmacology , Copper/therapeutic use , Tumor Microenvironment , Biomimetics , Disulfiram/pharmacology , Ditiocarb/pharmacology , Ditiocarb/therapeutic use , Neoplasms/drug therapy , Neoplasms/pathologyABSTRACT
BACKGROUND: BaeS/BaeR is a two-component system of Escherichia coli that controls the expression of porins and efflux pumps. Its role in beta-lactam resistance is limited. OBJECTIVES: To study the role of baeS/baeR two-component system in temocillin resistance in E. coli. METHODS: E. coli strain BW25113 and single-gene deletion mutants related to two-component systems were collected from the KEIO collection. Double-gen deletion mutants were generated. Temocillin-resistant mutant frequencies were determined at 32â mg/L. E. coli BW25113 mutants were selected by selective pressure from serial passages. Biological costs were analysed by growth curves. Genomes of the generated mutants were sequenced. The expression level of the mdtA, mdtB, mdtC, acrD and tolC in the ΔbaeS mutant was determined by RT-PCR (with/without temocillin exposure). RESULTS: The frequency of temocillin mutants ranged from 2.12 × 10-8 to 4.51 × 10-8 in single-porin mutants. No mutants were recovered from E. coli BW25113 (>10-9). Selection of temocillin-resistant variants by serial passage yielded mutants up to 128â mg/L. Mutations were found in the baeS gene. Temocillin MICs ranged from 4 to 32â mg/L (highest MICs for ΔbaeS and ΔompR). The efflux pumps mdtA, mdtB, mdtC and acrD pumps were overexpressed 3-10-fold in the presence of temocillin in ΔbaeS compared to control. CONCLUSIONS: Mutations in the sensor histidine kinase, baeS, may be involved in temocillin resistance through the expression of the efflux pumps mdtABC and acrD. In addition, the low mutation rate may be a good predictor of temocillin activity.
Subject(s)
Cadaverine/analogs & derivatives , Escherichia coli Proteins , Escherichia coli , Penicillins , Escherichia coli/genetics , Biological Transport , Trans-Activators , Escherichia coli Proteins/geneticsABSTRACT
Ten Gouda cheese wheels with an age of 31 weeks from six different batch productions were affected by a crack defect and displayed an unpleasant off-flavor. To unravel the causes of these defects, the concentrations of free amino acids, other organic acids, volatile organic compounds, and biogenic amines were quantified in zones around the cracks and in zones without cracks, and compared with those of similar Gouda cheeses without crack defect. The Gouda cheeses with cracks had a significantly different metabolome. The production of the non-proteinogenic amino acid γ-aminobutyric acid (GABA) could be unraveled as the key mechanism leading to crack formation, although the production of the biogenic amines cadaverine and putrescine contributed as well. High-throughput amplicon sequencing of the full-length 16S rRNA gene based on whole-community DNA revealed the presence of Loigolactobacillus rennini and Tetragenococcus halophilus as most abundant non-starter lactic acid bacteria in the zones with cracks. Shotgun metagenomic sequencing allowed to obtain a metagenome-assembled genome of both Loil. rennini and T. halophilus. However, only Loil. rennini contained genes necessary for the production of GABA, cadaverine, and putrescine. Metagenetics further revealed the brine and the rennet used during cheese manufacturing as the most plausible inoculation sources of both Loil. rennini and T. halophilus.IMPORTANCECrack defects in Gouda cheeses are still poorly understood, although they can lead to major economic losses in cheese companies. In this study, the bacterial cause of a crack defect in Gouda cheeses was identified, and the pathways involved in the crack formation were unraveled. Moreover, possible contamination sources were identified. The brine bath might be a major source of bacteria with the potential to deteriorate cheese quality, which suggests that cheese producers should regularly investigate the quality and microbial composition of their brines. This study illustrated how a multiphasic approach can understand and mitigate problems in a cheese company.
Subject(s)
Carboxy-Lyases , Cheese , Lactobacillales , Lactobacillus , Salts , Lactobacillales/genetics , Cheese/microbiology , RNA, Ribosomal, 16S/genetics , Cadaverine , Putrescine , Bacteria/genetics , gamma-Aminobutyric Acid , Lactic Acid , Food MicrobiologyABSTRACT
Necro-leachate, a liquid released during cadaveric decomposition, is considered the main culprit for impacts on cemetery environments. The biogenic amines cadaverine and putrescine make up part of the composition of necro-leachate and have a certain toxicity to different organisms. Springtails are among the most used bioindicators to assess the impacts of soil contaminants. As there are no data on the acute and chronic toxicity of springtails exposed to cadaverine and putrescine, the objective of this study was to evaluate the toxic potential of both amines, under the behavioral effect of avoidance and reproduction in the species Folsomia candida. Springtails were exposed to soils contaminated with different concentrations of cadaverine and putrescine, and different mixtures of these amines. To evaluate the avoidance and reproduction test, the individuals were exposed for periods of 48 h and 28 days, respectively. The results obtained in the avoidance test showed that springtails exhibited a preference for the treated soil in both isolated and mixed treatments. The chronic evaluation assays showed that the reproduction was affected, particularly in the treatments with combined amines, resulting in a reduction in the total number of juveniles. From the results, it is possible to infer that the methods applied in this research have provided information that will contribute to a better understanding of the toxicity of putrefactive biogenic amines, since there exist few ecotoxicological studies carried out with these amines, and especially with those from cemetery environments.
Subject(s)
Arthropods , Putrescine , Humans , Animals , Cadaverine , Environmental Monitoring , Cadaver , Biogenic Amines/toxicity , SoilABSTRACT
Amine determination is crucial to our daily life, including the prevention of pollution, the treatment of certain disorders, and the evaluation of food quality. Herein, a mixed-linkage donor-acceptor covalent organic framework (named DSE-COF) was first constructed by the polymerization between 2,4-dihydroxybenzene-1,3,5-tricarbaldehyde (DTA) and 4,4'-(benzo[c][1,2,5]selenadiazole-4,7-diyl)dianiline (SEZ). DSE-COF displayed superior turn-on fluorescent responses to primary, secondary, and tertiary aliphatic amines, such as cadaverine, isopropylamine, sec-butylamine, cyclohexylamine, hexamethylenediamine, di-n-butylamine, and triethylamine in absolute acetonitrile than other organic species. Further experiments and theoretical calculations demonstrated that the combination of intramolecular charge transfer (ICT) and photoinduced electron transfer (PET) effects between the DSE-COF and aliphatic amines resulted in enhanced fluorescence. Credibly, DSE-COF can quantitatively detect cadaverine content in actual pork samples with satisfactory results. In addition, DSE-COF-based test papers could rapidly monitor cadaverine from real pork samples, manifesting the potential application of COFs in food quality inspection.
Subject(s)
Cockayne Syndrome , Metal-Organic Frameworks , Humans , Cadaverine , Amines , Cyclohexylamines , Coloring AgentsABSTRACT
Escherichia coli and Shigella spp. are closely related, making it crucial to accurately identify them for disease control and prevention. In this study, we utilized MALDI-TOF MS to identify characteristic peaks of decarboxylation products of lysine and ornithine to distinguish between E. coli and Shigella spp. Our findings indicate that the peak at m/z 103.12 ± 0.1 of the product cadaverine from lysine decarboxylase is unique to E. coli, while all Shigella species lack the m/z 103.12 ± 0.1 peak. However, S. sonnei and S. boydii serotype C13 exhibit a specific peak at m/z 89.10 ± 0.1, which is the product of putrescine from ornithine decarboxylase. We were able to correctly identify 97.06% (132 of 136) of E. coli and Shigella isolates and 100% (8 of 8) of S. sonnei isolates using this biochemical-based MALDI-TOF MS detection system. This technology is advantageous for its high-throughput, high quality, and ease of operation, and is of significant value for the diagnosis of E. coli and Shigella-related diseases.
Subject(s)
Escherichia coli , Shigella , Escherichia coli/chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Shigella/chemistry , Cadaverine , PutrescineABSTRACT
Pseudomonas putida strain U can be grown using, as sole carbon sources, the biogenic amines putrescine or cadaverine, as well as their catabolic intermediates, ɣ-aminobutyrate or δ-aminovalerate, respectively. Several paralogs for the genes that encode some of the activities involved in the catabolism of these compounds, such as a putrescine-pyruvate aminotransferase (spuC1 and spuC2 genes) and a ɣ-aminobutyrate aminotransferase (gabT1 and gabT2 genes) have been identified in this bacterium. When the expression pattern of these genes is analyzed by qPCR, it is drastically conditioned by supplying the carbon sources. Thus, spuC1 is upregulated by putrescine, whereas spuC2 seems to be exclusively induced by cadaverine. However, gabT1 increases its expression in response to different polyamines or aminated catabolic derivatives from them (i.e., ɣ-aminobutyrate or δ-aminovalerate), although gabT2 does not change its expression level concerning no-amine unrelated carbon sources (citrate). These results reveal differences between the mechanisms proposed for polyamine catabolism in P. aeruginosa and Escherichia coli concerning P. putida strain U, as well as allow a deeper understanding of the enzymatic systems used by this last strain during polyamine metabolism.
Subject(s)
Pseudomonas putida , Putrescine , Cadaverine/metabolism , Putrescine/metabolism , Putrescine/pharmacology , Pseudomonas putida/genetics , Pseudomonas putida/metabolism , Polyamines/metabolism , Pseudomonas aeruginosa/genetics , Escherichia coli/genetics , Aminobutyrates/metabolism , Carbon/metabolism , Gene ExpressionABSTRACT
Cadaverine (Cad), which has an independent synthesis pathway compared to other polyamine (PA) types, contributes to the health of plants by regulating plant growth and development, abiotic stress tolerance and antioxidant defense mechanisms. In this work, experiments were carried out to understand the effects of exogenous Cad (10 µM) application under drought stress (%22 PEG 6000) and without stress on cell cycle, total protein content, endogenous PA levels, and biochemical enzyme activities in barley (Hordeum vulgare cv. Burakbey) considering the potential of Cad to stimulate the drought-related tolerance system. Cad application in a stress-free environment showed an effect almost like low-impact drought stress, causing changes in all parameters examined compared to samples grown in distilled water environment (Control). The results clearly show that Cad applied against the negative effects of drought stress on all parameters creates a drought resistance mechanism of the plant. Accordingly, Cad applied together with drought stress increased the density of cells in the cell cycle (G1-S and S-G2 phases) and the amount of endogenous (spermidine 10% and spermine 40%) PAs. In addition, while superoxide dismutase (SOD) (5%), (CAT) (55%) and ascorbate peroxidase (APX) (18%) enzyme levels increased, a stress response mechanism occurred due to the decrease in total protein content (20%) and malondialdehyde (MDA) (80%). As a result, exogenous application of 10 µM Cad showed that it reduced the negative effects of drought stress on endogenous PA amounts, cell division and biochemical activities in barley.
Subject(s)
Hordeum , Polyamines , Polyamines/metabolism , Hordeum/metabolism , Cadaverine/pharmacology , Cadaverine/metabolism , Seedlings , Droughts , Antioxidants/metabolism , Cell Cycle , Cell DivisionABSTRACT
Chlamydomonas reinhardtii (C. reinhardtii) is a single-cell green alga that can be easily genetically manipulated. With its favorable characteristics of rapid growth, low cost, non-toxicity, and the ability for post-translational protein modification, C. reinhardtii has emerged as an attractive option for the biosynthesis of various valuable products. To enhance the expression level of exogenous genes and overcome the silencing of foreign genes by C. reinhardtii, synthetic promoters such as the chimeric promoter AR have been constructed and evaluated. In this study, a synthetic promoter GA was constructed by hybridizing core fragments from the natural promoters of the acyl carrier protein gene (ACP2) and the glutamate dehydrogenase gene (GDH2). The GA promoter exhibited a significant increase (7 times) in expressing GUS, over the AR promoter as positive control. The GA promoter also displayed a strong responsiveness to blue light (BL), where the GUS expression was doubled compared to the white light (WL) condition. The ability of the GA promoter was further tested in the expression of another exogenous cadA gene, responsible for catalyzing the decarboxylation of lysine to produce cadaverine. The cadaverine yield driven by the GA promoter was increased by 1-2 times under WL and 2-3 times under BL as compared to the AR promoter. This study obtained, for the first time, a blue light-responsive GDH2 minimal fragment in C. reinhardtii, which delivered a doubling effect under BL when used alone or in hybrid. Together with the strong GA synthetic promoter, this study offered useful tools of synthetic biology to the algal biotechnology field.
Subject(s)
Chlamydomonas reinhardtii , Chlamydomonas reinhardtii/genetics , Chlamydomonas reinhardtii/metabolism , Cadaverine/metabolism , Promoter Regions, Genetic , Biotechnology , LightABSTRACT
Aldehyde-functionalized cellulose (AFC) was prepared by oxidizing cellulose with sodium metaperiodate. The reaction was characterized by Schiff's test, FT-IR, and UV-vis study. AFC was evaluated as a reactive sorbent for controlling polyamine-based odor from chronic wounds, and its performance was compared with charcoal, one of the most widely utilized odor-control sorbents through physisorption. Cadaverine was used as the model odor molecule. A liquid chromatography/mass spectrometry (LC/MS) method was established to quantify the compound. AFC was found to rapidly react with cadaverine through the Schiff-base reaction, which was confirmed by FT-IR, visual observation, CHN elemental analysis, and the ninhydrin test. The sorption and desorption behaviors of cadaverine onto AFC were quantified. With clinic-relevant cadaverine concentrations, AFC demonstrated much better sorption performance than charcoal. At even higher cadaverine concentrations charcoal showed higher sorption capacity, probably due to its high surface area. On the other hand, in desorption studies, AFC retained much more of the sorbed cadaverine than charcoal. When AFC and charcoal were combined, the pair demonstrated excellent sorption and desorption behaviors. The XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) assay confirmed that AFC has very good in vitro biocompatibility. These results suggest that AFC-based reactive sorption can be a new strategy to control odors associated with chronic wounds for improved healthcare.