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Cholestatic liver disease (CLD) is a common liver disorder with limited treatment options. Here, we demonstrate that zinc (Zn) supplementation can alter the gut microbiome to mitigate cholestatic liver injury. Oral Zn altered the microbiota of mice and humans (this study was registered at clinicaltrials.gov [NCT05597137]), increasing the abundance of Blautia producta (B. producta) and promoting the generation of p-coumaric acid. Additionally, p-coumaric acid concentrations were negatively correlated with liver injury parameters in CLD patients. In mice, the protective effects of Zn were partially mediated by p-coumaric acid, which directly bound to nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX2) and suppressed the production of reactive oxygen species (ROS) in hepatocytes, thus preventing hepatocyte cell death and liver damage. Additionally, knocking out the histidine ammonia-lyase, which catalyzes the conversion of tyrosine to p-coumaric acid in B. producta , blunted the protective effects of Zn. These findings highlight a host-microbiota interaction that is stimulated by Zn supplementation, providing potential benefits for CLD. [Display omitted] • Zn administration modulates the gut microbiota and mitigates CLD • Zn boosts the generation of p-coumaric acid by B. producta • p-coumaric acid is catalyzed by the histidine ammonia-lyase in B. producta • p-coumaric acid ameliorates bile-acid-induced hepatocyte pyroptosis Li et al. show that zinc impacts the gut microbiome and specifically Blautia producta (B. producta) with ameliorative effects for cholestatic liver disease. Zn enhances p-coumaric acid production by B. producta , which reduces bile-acid-induced hepatocyte pyroptosis by inhibiting the NOX2-ROS-GSDME axis, thus reducing liver injury. [ABSTRACT FROM AUTHOR]

In this study, we determine the chemical composition, bioactive compounds profile, antioxidant capacity and antidiabetic potential of two cactus fruits. Pilosocereus gounellei had the highest contents of ash (6.84%), lipid (7.79%), maltose (1.02 mg g−1), soluble fiber (10.85%) and minerals such as Mg. Thirteen volatile compounds (alcohols, terpenes, aldehydes and esters) were found in the fruits. Although higher amounts of quinic acid and quercetrin were found in Cereus Jamacaru, Pilosocereus gounellei had a higher content of p-coumaric acid and rutin. Regarding the functional potential, Cereus Jamacaru had the highest content of total phenolics (65.17 GAE g−1), total flavonoids (26.06 mg EC g−1), antioxidant capacity (28.67; 49.86; 158.79 µM TE g−1 for DPPH, ABTS and ORAC, respectively) and highest inhibition for α-amylase (84.84 ± 0.41%) and protein glycation. In view of their nutritional and bioactive characteristics, these species have great potential to be used in preparations with functional claims. [ABSTRACT FROM AUTHOR]

To investigate the effect of p‐coumaric acid (p‐CA) on postharvest cherry tomatoes, breaker stage fruits were treated with p‐CA to analyze the physiological metabolism during storage. The results showed that exogenous p‐CA treatment improved the sensory quality of tomato fruits. Transcriptomics results indicated that 782 genes (339 up‐regulated and 443 down‐regulated) were differentially expressed between p‐CA treated and control fruits. Results suggested that p‐CA treatment regulated the synthesis of phenolic compounds and inhibited the fruit ripening through the pathways of mangiferic acid biosynthesis and phenylalanine metabolism. Key enzymes activities of the phenylpropane metabolic pathway of tomato fruits were increased, including phenylalanine ammonia lyase (PAL), cinnamic acid 4‐hydroxylase (C4H), and 4‐coumarate‐CoA ligase (4CL). In addition, the total phenols content, flavonoids content and antioxidant capacities of tomato fruits were improved with p‐CA treatment. Overall, these findings showed that p‐CA treatment could be a potential strategy for fruit and vegetable preservation. [ABSTRACT FROM AUTHOR]

Natural alkylated hydroxy cinnamates (AHCs) isolated from medicinal plants and the thereby designed and synthesized cinnamides are derivatives of hydroxy cinnamic acids such as p-coumaric, sinapic, ferulic, and caffeic acids, which are naturally derived from human dietary sources. The pharmacological properties displayed by AHCs based on their inherent structure range include antioxidant, antimicrobial, antiplasmodial, anti-tyrosinase, Alzheimer’s and Parkinson’s disease therapy, anticancer therapy, metabolic disease therapy, and biopesticides, which have not been reviewed together. Based on their inherent antioxidant, antimicrobial, and UV absorption and their structure–activity relationships, these cinnamyl esters and amides can be used for food preservation in emulsions and oils, as sun-protective components of skin care formulations, and in many other multifunctional applications. In conclusion, the fine-tuning of the structural features such as the type of hydroxy cinnamic acid used, the length of alkyl chains for variable lipophilicity, conversion from cinnamic to propanoic for antioxidants, the increase in methoxy or the change to amino groups to increase the molar absorption coefficient and loss of absorption values, the substitution by halides or amino groups for potent biopesticides, and conversion from esters to amide bonds leads to different AHCs for biomedical, cosmetic, and agriculture applications as an emerging field of investigation that can overall provide natural, safe, biodegradable, and sustainable molecules.

Genetic logic gates can be employed in metabolic engineering and synthetic biology to regulate gene expression based on diverse inputs. Design of tunable genetic logic gates with versatile dynamic performance is essential for expanding the usability of these toolsets. Here, using the p-coumaric acid biosensor system as a proof-of-concept, we initially investigated the parameters influencing the buffer (BUF) genetic logic gates. Subsequently, integrating binding sequences from the p-coumaric acid biosensor system and tetR or lacI regulation systems into a constitutive promoter yielded AND genetic logic gates. Additionally, characterized antisense RNAs (asRNAs) or single guide RNAs (sgRNAs) with various repression efficiencies were combined with BUF gates to construct a suite of p-coumaric acid-triggered NOT genetic logic gates. Finally, the designed BUF and NOT gates were combined to construct bifunctional genetic circuits that were subjected to orthogonality evaluation. The genetic logic gates established in this study can serve as valuable tools in future applications of metabolic engineering and synthetic biology.

Summary: This study focused on the fabrication of zein‐based coatings functionalised by two phenolic acids, caffeic acid (CA) and p‐coumaric acid (pCA), using electrospinning. The electrospun fibres were fabricated with three different concentrations (5%, 10% and 20% w/w) of CA and pCA individually. The average fibre diameter (AFD) increased due to the addition of phenolic acids. Thermogravimetric analysis (TGA) revealed that degradation temperatures of the zein electrospun fibres were not significantly affected (P > 0.05) after the incorporation of CA and pCA. Meanwhile, interactions between zein and the phenolic acids were indicated by surface characterisation. CA‐loaded zein electrospun fibres exhibited increasing antioxidant activity with increasing CA concentration. Both phenolic acid‐loaded zein electrospun fibres displayed favourable antibacterial activities against S. aureus and E. coli foodborne pathogens. Overall, zein electrospun fibres with 20% w/w CA demonstrated the most desirable properties for potential active food packaging application. [ABSTRACT FROM AUTHOR]

Background: Hyperpigmentation, frequently triggered by an excessive production of melanin, is a common issue within the realms of dermatology and cosmetology. In addition to regulating tyrosinase activity, the autophagy process plays a role in melanosome turnover, contributing to pigmentation control. p-Coumaric acid (PCA), a dietary phenolic compound with antioxidant and anti-inflammatory properties, was investigated for its dual suppressive effects on melanin production induced by alpha-melanocyte-stimulating hormone (α-MSH) and autophagy inhibitors in B16F10 cells. Results: PCA (25–100 µg/mL) serves as a potent in vitro inhibitor of tyrosinase activity. In addition, PCA can effectively mitigate the upregulation of tyrosinase gene expression (P < 0.01) and its cellular activities induced by α-MSH. In contrast to early-stage autophagy inhibitors like SBI0206965 (SBI) and spautin-1, treatment with 50 µM of chloroquine (CQ) and 20 nM of bafilomycin A1 (BFA), both of which inhibit the late stages of the autophagic process, results in an increase in melanin content within B16F10 cells, independent of cellular tyrosinase activity. Furthermore, PCA treatment could protect cells against CQ and BFA-induced lysosomal damage, ultimately leading to the promotion of autolysosome formation and the activation of the autophagic process, which results in melanin degradation. Conclusions: In summary, PCA exhibits dual suppressive effects on melanogenesis via inhibiting tyrosinase activity and melanin accumulation caused by lysosomal dysfunction. These effects offer an enhanced opportunity for the development of a safe and effective anti-melanogenesis agent. [ABSTRACT FROM AUTHOR]

Introduction: Applying biochar (BC) to reduce toxic substance accumulation in soil, either through direct adsorption or modulation of the microbial community, has received considerable attention. However, a knowledge gap exists regarding how BC regulates microbial community structure and functions to mitigate toxic substance accumulation. Methods: We previously identified p-coumaric acid (p-CA) as a representative autotoxin in tobacco rhizosphere soil. On this basis, this study simulated a soil environment with p-CA accumulation to investigate the impacts of BC on p-CA, soil physicochemical properties, and microbial community structure and function. Results: The results showed that p-CA could be directly adsorbed onto BC, which followed the pseudo-second-order kinetic model (R2 = 0.996). A pot experiment revealed that BC significantly reduced soil p-CA, altered soil microbial composition, and enhanced bacterial community diversity. A weighted correlation network analysis showed a close association between taxon 1 in the microbial network and p-CA, suggesting a pivotal role for this taxon in reducing p-CA, with Devosia and Nocardioides identified as potential key contributors to this process. The prediction of possible keystone taxa functions showed that BC increased the relative abundances of aromatic compound degraders. Mantel tests indicated that soil organic matter exerted the greatest influence on keystone taxa functions and hub genera. Discussion: These findings suggest that BC may either directly chemisorb p-CA or indirectly facilitate p-CA degradation by regulating the functioning of keystone taxa. The results of this study provide a novel perspective for further investigation of the mechanisms through which BC reduces the accumulation of toxic substances in soil. [ABSTRACT FROM AUTHOR]

The economic competitiveness of 2G-bioethanol technology should improve through the improvement of the sugar release and the valorization of by-products, especially lignin. Thus, an integrated scheme with corncob was developed to produce ethanol using low dosages of cellulases and value-added products from the semi-simultaneous saccharification and fermentation (SSSF) residue. Enzymatic hydrolysis and SSSF of acid pretreated corncob (< 20 mesh and > 20 mesh) were carried out under cellulase dosages of 5, 10, and 15 FPU/g in the absence and presence of polyethylene glycol 1500 (PEG 1500). The SSSF residue was used to obtain lignosulfonate via sulfomethylation reaction and phenolic acids via alkaline hydrolysis using 4% (w/v) sodium hydroxide and 0–5% (v/v) hydrogen peroxide. Pretreated corncob < 20 mesh allowed the reduction of cellulase dosage to 5 FPU/g without compromising sugar release. The addition of PEG 1500 boosted sugar release, reaching 56.73 g/L glucose under 20% (w/v) solids. The maximum ethanol production of 31.64 g/L was obtained using 5 FPU/g cellulases, 2% (w/w) PEG 1500, and 20% (w/v) solids (gradual addition). FTIR confirmed the preparation of lignosulfonate from SSSF residue, and the surfactant showed good stabilization performance in oil/water systems (emulsification index≈30%). High yields of p-coumaric acid (8045.3 mg/100 g) and ferulic acid (1429.4 mg/100 g) were obtained in alkaline hydrolysis with 5% (v/v) hydrogen peroxide. Based on these findings, corncob is versatile and can create a biorefinery with high economic potential. [ABSTRACT FROM AUTHOR]

Ferulic acid and p-coumaric acid, cinnamic derivatives of phenolic acid, have antibacterial, prooxidant, and antioxidant effects. In this study ferulic acid and p-coumaric acid amide derivatives were investigated for their antibacterial and antioxidant properties are described in this communication. The most effective conjugates against B subtilis were 5b (IC50: 215 ​± ​1.3 ​μM) and 4d (IC50: 336 ​± ​2.7 ​μM) and against P. aeruginosa were 4b (IC50: 365 ​± ​2.8 ​μM) and 5b (IC50: 341 ​± ​3.6 ​μM), whereas the none of conjugates were more effective against E. coli than reference Kanamycin. Conjugates 5b was the most effective against B subtilis of all the synthesized conjugates, with IC50 values of (IC50: 215 ​± ​1.3 ​μM). The free radical scavenging capacity of each compound was determined using the DPPH and ABTS assays. Conjugates 4b (IC50: 53 ​± ​3.6 ​μM), 4c (IC50: 58 ​± ​1.3 ​μM), 4d (IC50: 57 ​± ​2.5 ​μM), 5b (IC50: 29 ​± ​1.5 ​μM) and 4a (IC50: 56 ​± ​4.3 ​μM) have greater antioxidant capacity than ferulic acid and ascorbic acid in the DPPH assay. Whereas in the ABTS assay, compounds 4b (IC50: 7 ​± ​1.8 ​μM), 5b (IC50: 5 ​± ​0.7 ​μM), 4a (IC50: 9 ​± ​3.2 ​μM), 4g (IC50: 7 ​± ​2.3 ​μM), and 5a (IC50: 8 ​± ​4.3 ​μM) showed more antioxidant activity than ferulic acid, p-coumaric acid and ascorbic acid. Thus, a large library of compounds derived from bile acid can be easily synthesized for extensive structure-activity relationship studies in order to identify the most appropriate antibacterial and antioxidant agents.

Stone-fruit liqueurs contain high contents of the carcinogen ethyl carbamate (EC). In this study, we investigated the effect of plum fruit extract and single antioxidants present in plum fruit extracts on the reduction in the EC content during the macerating process in a plum liqueur model system and authentic plum liqueur. 30% ethanol model plum liqueur treated with 0.2% plum extract showed the lowest EC content with 55% reduction rate after the macerating process compared to the content in the control. Interestingly, neither 0.1% ascorbic acid nor 0.1% p-coumaric acid lowered the EC contents in the model liqueur, while they decreased the EC contents in authentic plum liqueur. This was possibly attributed to the synergistic effect of the plum fruit phenolics with the ascorbic acid and p-coumaric acid antioxidants. Thus, plum extracts can be applied to plum liqueurs to reduce the rate of EC formation. [ABSTRACT FROM AUTHOR]

P‐coumaric acid (p‐CA), a pant metabolite with antioxidant and anti‐inflammatory activity, is extensively utilized in biomedicine, food, and cosmetics industry. In this study, a synthetic pathway (PAL) for p‐CA was designed, integrating three enzymes (AtPAL2, AtC4H, AtATR2) into a higher l‐phenylalanine‐producing strain Escherichia coli PHE05. However, the lower soluble expression and activity of AtC4H in the PAL pathway was a bottleneck for increasing p‐CA titers. To overcome this limitation, the soluble expression of AtC4H was enhanced through N‐terminal modifications. And an optimal mutant, AtC4HL373T/G211H, which exhibited a 4.3‐fold higher kcat/Km value compared to the wild type, was developed. In addition, metabolic engineering strategies were employed to increase the intracellular NADPH pool. Overexpression of ppnk in engineered E. coli PHCA20 led to a 13.9‐folds, 1.3‐folds, and 29.1% in NADPH content, the NADPH/NADP+ ratio and p‐CA titer, respectively. These optimizations significantly enhance p‐CA production, in a 5‐L fermenter using fed‐batch fermentation, the p‐CA titer, yield and productivity of engineered strain E. coli PHCA20 were 3.09 g/L, 20.01 mg/g glucose, and 49.05 mg/L/h, respectively. The results presented here provide a novel way to efficiently produce the plant metabolites using an industrial strain. [ABSTRACT FROM AUTHOR]

Potassium bromate (KBrO3) is a common dietary additive, pharmaceutical ingredient, and significant by‐product of water disinfection. p‐coumaric acid (PCA) is a naturally occurring nutritional polyphenolic molecule with anti‐inflammatory and antioxidant activities. The goal of the current investigation was to examine the protective effects of p‐coumaric acid against the liver damage caused by KBrO3. The five groups of animals‐control, KBrO3 (100 mg/kg bw), treatment with KBrO3 along with Silymarin (100 mg/kg bw), KBrO3, followed by PCA (100 mg/bw, and 200 mg/kg bw) were randomly assigned to the animals. Mice were slaughtered, and blood and liver tissues were taken for assessment of the serum biochemical analysis for markers of liver function (alanine transaminase, aspartate transaminase, alkaline phosphatase, albumin, and protein), lipid markers and antioxidant markers (TBARS), glutathione peroxidase [GSH‐Px], glutathione (GSH), and markers of hepatic oxidative stress (CAT), (SOD), as well as histological H&E stain, immunohistochemical stain iNOS, and COX‐2 as markers of inflammatory cytokines. PCA protects against acute liver failure by preventing the augmentation of blood biochemical markers and lipid profiles. In mice liver tissues, KBrO3 increases lipid indicators and depletes antioxidants, leading to an increase in JNK, ERK, and p38 phosphorylation. Additionally, PCA inhibited the production of pro‐inflammatory cytokines and reduced the histological alterations in KBrO3‐induced hepatotoxicity. Notably, PCA effectively mitigated KBrO3‐induced hepatic damage by obstructing the TNF‐α/NF‐kB‐mediated inflammatory process signaling system. Additionally, in KBrO3‐induced mice, PCA increased the intensities of hepatic glutathione (GSH), SOD, GSH‐Px, catalase, and GSH activities. Collectively, we demonstrate the molecular evidence that PCA eliminated cellular inflammatory conditions, mitochondrial oxidative stress, and the TNF‐α/NF‐κB signaling process, thereby preventing KBrO3‐induced hepatocyte damage. Significant statement: Based on our study's biochemical and histological results, we have definitively proven that p‐coumaric acid (PCA) shields the liver in the experimental model against oxidative damage caused by potassium bromate (KBrO3). PCA also prevents oxidative stress. Additionally, in KBrO3‐induced mice, PCA increased the intensities of hepatic glutathione, superoxide dismutase, glutathione peroxidase, catalase, and glutathione reductase activities. Notably, PCA effectively mitigated KBrO3‐induced hepatic damage by obstructing the TNF‐α/NF‐kB‐initiated inflammatory response signaling pathway. Collectively, we provide molecular evidence that PCA reduced cellular inflammation, mitochondrial oxidative stress, and the TNF‐α/NF‐κB signaling cascade to prevent KBrO3‐induced hepatocyte damage. In a nutshell, the results of this investigation suggest that PCA therapy may be helpful in reducing oxidative stress in liver damage brought on by KBrO3. [ABSTRACT FROM AUTHOR]

Copyright of Farabi Medical Journal is the property of Farabi Medical Journal and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Probiotics are defined as “live microorganisms that provide health benefits to the host when administered in adequate amounts.” Probiotics have beneficial effects on human health, including antibacterial activity against intestinal pathogens, regulation of blood cholesterol levels, reduction of colitis and inflammation incidence, regulation of the immune system, and prevention of colon cancer. In addition to probiotic bacteria, some phenolic compounds found in foods we consume (both food and beverages) have positive effects on human health. p-coumaric acid (p-CA) is one of the most abundant phenolic compounds in nature and human diet. The interactions between these two different food components (phenolics and probiotics), resulting in more beneficial combinations called synbiotics, are not well understood in terms of how they will affect the gut microbiota by promoting the probiotic properties and growth of probiotic bacteria. Thus, this study aimed to investigate synbiotic relationship between p-CA and Lactobacillus acidophilus LA-5 (LA-5), Lacticaseibacillus rhamnosus GG (LGG). Probiotic bacteria were grown in the presence of p-CA at different concentrations, and the effects of p-CA on probiotic properties, as well as its in vitro effects on AChE and BChE activities, were investigated. Additionally, Surface analysis was conducted using FTIR. The results showed that treatment with p-CA at different concentrations did not exhibit any inhibitory effect on the growth kinetics of LA-5 and LGG probiotic bacteria. Additionally, both probiotic bacteria demonstrated high levels of antibacterial properties. It showed that it increased the auto-aggregation of both probiotics. While p-CA increased co-aggregation of LA-5 and LGG against Escherichia coli, it decreased co-aggregation against Staphylococcus aureus. Probiotics grown with p-CA were more resistant to pepsin. While p-CA increased the resistance of LA-5 to bile salt, it decreased the resistance of LGG. The combinations of bacteria and p-CA efficiently suppressed AChE and BChE with inhibition (%) 11.04–68.43 and 13.20-65.72, respectively. Furthermore, surface analysis was conducted using FTIR to investigate the interaction of p-coumaric acid with LA-5 and LGG, and changes in cell components on the bacterial surface were analyzed. The results, recorded in range of 4000 –600 cm-1 with resolution of 4 cm-1, demonstrated that p-CA significantly affected only the phosphate/CH ratio for both bacteria. These results indicate the addition of p-CA to the probiotic growth may enhance the probiotic properties of bacteria. [ABSTRACT FROM AUTHOR]

This study investigated the efficacy of p-coumaric acid (p-CA), a natural phenolic compound, on traumatic brain injury (TBI)-induced spatial cognitive deficits and the possible involved mechanisms. Rats received p-CA (100 mg/kg, orally) 30 min after diffuse TBI induction. Spatial memory, neuroplasticity, oxidative stress, and histological alterations were evaluated at scheduled time points. TBI reduced spatial memory capacity on days 1, 3, and 7 in the water-maze task, which was associated with suppressed hippocampal long-term potentiation (LTP) at the perforant path-dentate gyrus (PP-DG) synapses. These deficits were accompanied by the inhibition of the activities of antioxidant enzymes and the promotion of lipid peroxidation in the hippocampal and cerebral cortex areas. Moreover, TBI resulted in neuronal loss in the DG. Interestingly, p-CA treatment ameliorated all the above-mentioned TBI outcomes. The findings demonstrate that p-CA alleviates TBI-induced spatial cognitive impairment, possibly by mitigating hippocampal synaptic dysfunction, modulating oxidative-antioxidative status, and preventing neuronal loss.

There is a balance between DM yield and feed value when choosing types of grasses on a farm depending on the acreages of farmland and types of ruminants to be fed. Therefore, optimisation of the harvest strategy for grass silage is important for profitable dairy farming. Tall fescue has high DM yield and can replace traditional grasses, such as timothy, in Northern Europe in a changing climate as it has been shown to be more drought tolerant. As differences in climate responses previously have been related to differences in cell wall structure between grass species and, consequently, in digestibility, it is highly relevant to compare these species at similar maturity stages and to investigate if a very early harvest date will diminish potential differences between the species. This study evaluated the effects of harvest date and forage species on the concentration of hydroxycinnamic acids in silages and its relationship to feed efficiency of dairy cows. Tall fescue and timothy were harvested at very early date on May 25 or at early date on May 31 in the spring growth cycle. Forty lactating dairy cows were used in a block design. Cows received 1 of 4 treatments: (1) tall fescue harvested at very early date, (2) timothy harvested at very early date, (3) tall fescue harvested at early date, and (4) timothy harvested at early date. Diets were formulated to have the same forage-to-concentrate ratio (49:51 on DM basis). Tall fescue silages showed greater concentrations of DM, ash, and CP than timothy silages. Grasses harvested at early date showed greater concentrations of NDF, ADL, and cell wall than grasses harvested at very early date. Tall fescue silages showed greater concentration of p-coumaric acid and lower in vitro organic matter digestibility (IVOMD) compared to timothy silages. Milk production and composition were not affected by treatments but cows fed tall fescue-based diets showed lower milk protein yield and greater milk urea nitrogen than when timothy-based diets were fed. Furthermore, cows receiving timothy-based diets showed greater feed efficiency compared to cows receiving tall fescue-based diets. Thus, the lower concentration of p-coumaric acid and the higher IVOMD was associated with greater feed efficiency of cows fed timothy-based diets compared to tall fescue-based diets.

With rapid advances in the development of metabolic pathways and synthetic biology toolkits, a persisting challenge in microbial bioproduction is how to optimally rewire metabolic fluxes and accelerate the concomitant high-throughput phenotype screening. Here we developed a biosensor-assisted titratable CRISPRi high-throughput (BATCH) screening approach that combines a titratable mismatch CRISPR interference and a biosensor mediated screening for high-production phenotypes in Escherichia coli. We first developed a programmable mismatch CRISPRi that could afford multiple levels of interference efficacy with a one-pot sgRNA pool (a total of 16 variants for each target gene) harboring two consecutive random mismatches in the seed region of sgRNA spacers. The mismatch CRISPRi was demonstrated to enable almost a full range of gene knockdown when targeting different positions on genes. As a proof-of-principle demonstration of the BATCH screening system, we designed doubly mismatched sgRNA pools targeting 20 relevant genes in E. coli and optimized a PadR-based p-coumaric acid biosensor with broad dynamic range for the eGFP fluorescence guided high-production screening. Using sgRNA variants for the combinatorial knockdown of pfkA and ptsI, the p-coumaric acid titer was increased by 40.6% to o 1308.6 mg/l from glycerol in shake flasks. To further demonstrate the general applicability of the BATCH screening system, we recruited a HpdR-based butyrate biosensor that facilitated the screening of E. coli strains achieving 19.0% and 25.2% increase of butyrate titer in shake flasks with sgRNA variants targeting sucA and ldhA, respectively. This work reported the establishment of a plug-and-play approach that enables multilevel modulation of metabolic fluxes and high-throughput screening of high-production phenotypes.

Endogenous metabolic pathways in microbial cells are usually precisely controlled by sophisticated regulation networks. However, the lack of such regulations when introducing heterologous pathways in microbial hosts often causes unbalanced enzyme expression and carbon flux distribution, hindering the construction of highly efficient microbial biosynthesis systems. Here, using naringenin as the target compound, we developed an Autonomous Cascaded Artificial Dynamic (AutoCAD) regulation system to automatically coordinate the pathway expression and redirect carbon fluxes for enhanced naringenin production. The AutoCAD regulation system, consisting of both intermediate-based feedforward and product-based feedback control genetic circuits, resulted in a 16.5-fold increase in naringenin titer compared with the static control. Fed-batch fermentation using the strain with AutoCAD regulation further enhanced the naringenin titer to 277.2 mg/L. The AutoCAD regulation system, with intermediate-based feedforward control and product-triggered feedback control, provides a new paradigm of developing complicated cascade dynamic control to engineer heterologous pathways.

Abstract Background Phenylpropanoids are a large group of plant secondary metabolites with various biological functions, derived from aromatic amino acids. Cyanobacteria are promising host organisms for sustainable production of plant phenylpropanoids. We have previously engineered Synechocystis sp. PCC 6803 to produce trans-cinnamic acid (tCA) and p-coumaric acid (pCou), the first intermediates of phenylpropanoid pathway, by overexpression of phenylalanine- and tyrosine ammonia lyases. In this study, we aimed to enhance the production of the target compounds tCA and pCou in Synechocystis. Results We eliminated the 4-hydroxyphenylpyruvate dioxygenase (HPPD) activity, which is a competing pathway consuming tyrosine and, possibly, phenylalanine for tocopherol synthesis. Moreover, several genes of the terminal steps of the shikimate pathway were overexpressed alone or in operons, such as aromatic transaminases, feedback insensitive cyclohexadienyl dehydrogenase (TyrC) from Zymomonas mobilis and the chorismate mutase (CM) domain of the fused chorismate mutase/prephenate dehydratase enzyme from Escherichia coli. The obtained engineered strains demonstrated nearly 1.5 times enhanced tCA and pCou production when HPPD was knocked out compared to the parental production strains, accumulating 138 ± 3.5 mg L−1 of tCA and 72.3 ± 10.3 mg L−1 of pCou after seven days of photoautotrophic growth. However, there was no further improvement when any of the pathway genes were overexpressed. Finally, we used previously obtained AtPRM8 and TsPRM8 Synechocystis strains with deregulated shikimate pathway as a background for the overexpression of synthetic constructs with ppd knockout. Conclusions HPPD elimination enhances the tCA and pCou productivity to a similar extent. The use of PRM8 based strains as a background for overexpression of synthetic constructs, however, did not promote tCA and pCou titers, which indicates a tight regulation of the terminal steps of phenylalanine and tyrosine synthesis. This work contributes to establishing cyanobacteria as hosts for phenylpropanoid production.

Colistin-resistant Acinetobacter baumannii (COLR-Ab) is an opportunistic pathogen commonly associated with nosocomial infections, and it is difficult to treat with current antibiotics. Therefore, new antimicrobial agents need to be developed for treatment. Based on this information, we investigated the antimicrobial, antibiofilm, and combination activities of p-coumaric acid (p-CA), ferulic acid (FA), and p-methoxycinnamic acid (p-MCA) against five COLR-Ab isolates. p-CA, FA, and p-MCA exhibited antimicrobial activity against COLR-Ab isolates, with minimum inhibitory concentration (MIC) values in the range of 256–128 µg/mL, 1024–512 µg/mL, and 512–128 µg/mL, respectively. The combination effects of the compounds with colistin (COL) were evaluated using a checkerboard synergy test. The combinations exhibited a synergistic effect and caused a 128- to 16-fold decrease in COL MIC values. In addition, the biofilm production capacities of the COLR-Ab isolates and the antibiofilm activities of the compounds were determined using the microtitre plate-based crystal violet (CV) technique. The compounds showed effective antibiofilm activity against strong and moderate biofilm-producing isolates, inhibiting biofilm formation by 77.5% and 19.7%. Spectrometric measurements were used to examine the effect of compounds on membrane permeability; 1.9-, 1.66-, and 1.34-fold increases in absorbance values were observed at MIC concentrations of p-CA, FA, and p-MCA, respectively. Furthermore, morphological changes caused by the compounds in the isolate were observed using scanning electron microscopy (SEM) micrographs. According to the WST assay, the compounds did not show any statistically significant cytotoxic effect on the cells (p > 0.05). These findings indicate that p-CA, FA, and p-MCA may be potential new alternative candidates against resistant A. baumannii.

Caffeic acid (CA) is a phenolic acid compound widely used in pharmaceutical and food applications. However, the efficient synthesis of CA is usually limited by the resources of individual microbial platforms. Here, a cross-kingdom microbial consortium was developed to synthesize CA from sugarcane bagasse hydrolysate using Escherichia coli and Candida glycerinogenes as chassis. In the upstream E. coli module, shikimate accumulation was improved by intensifying the shikimate synthesis pathway and blocking shikimate metabolism to provide precursors for the downstream CA synthesis module. In the downstream C. glycerinogenes module, conversion of p-coumaric acid to CA was improved by increasing the supply of the cytoplasmic cofactor FAD(H2). Further, overexpression of ABC transporter-related genes promoted efflux of CA and enhanced strain resistance to CA, significantly increasing CA titer from 103.8 mg/L to 346.5 mg/L. Subsequently, optimization of the inoculation ratio of strains SA-Ec4 and CACg27 in this cross-kingdom microbial consortium resulted in an increase in CA titer to 871.9 mg/L, which was 151.6% higher compared to the monoculture strain CA-Cg27. Ultimately, 2311.6 and 1943.2 mg/L of CA were obtained by optimization of the co-culture system in a 5 L bioreactor using mixed sugar and sugarcane bagasse hydrolysate, respectively, with 17.2-fold and 14.6-fold enhancement compared to the starting strain. The cross-kingdom microbial consortium developed in this study provides a reference for the production of other aromatic compounds from inexpensive raw materials. [ABSTRACT FROM AUTHOR]

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This paper presents the results of a study of the effects of the lanthanide ions Ce3+, Pr3+, Nd3+ and Sm3+ on the electronic structure and antioxidant and biological (antimicrobial and cytotoxic) properties of p-coumaric acid (p-CAH2). Structural studies were conducted via spectroscopic methods (FTIR, ATR, UV). Thermal degradation studies of the complexes were performed. The results are presented in the form of TG, DTG and DSC curves. Antioxidant properties were determined via activity tests against DPPH, ABTS and OH radicals. The reducing ability was tested via CUPRAC assays. Minimum inhibitory concentrations (MICs) of the ligand and lanthanide complexes were determined on E. coli, B. subtilis and C. albicans microorganisms. The antimicrobial activity was also determined using the MTT assay. The results were presented as the relative cell viability of C. albicans, P. aeruginosa, E. coli and S. aureus compared to controls and expressed as percentages. In the obtained complexes in the solid phase, lanthanide ions coordinate three ligands in a bidentate chelating coordination mode through the carboxyl group of the acid. Spectroscopic analysis showed that lanthanide ions increase the aromaticity of the pi electron system of the ligand. Thermal analysis showed that the complexes are hydrated and have a higher thermal stability than the ligand. The products of thermal decomposition of the complexes are lanthanide oxides. In the aqueous phase, the metal combines with the ligand in a 1:1 molar ratio. Antioxidant activity tests showed that the complexes have a similar ability to remove free radicals. ABTS and DPPH tests showed that the complexes have twice the ability to neutralise radicals than the ligand, and a much higher ability to remove the hydroxyl radical. The abilities of the complexes and the free ligand to reduce Cu2+ ions in the CUPRAC test are at a similar level. Lanthanide complexes of p-coumaric acid are characterised by a higher antimicrobial capacity than the free ligand against Escherichia coli bacteria, Bacillus subtilis and Candida albicans fungi. [ABSTRACT FROM AUTHOR]

Novel p-coumaric acid microemulsion systems were developed to circumvent its absorption and bioavailability challenges. Simplex-lattice mixture design and machine learning methods were employed for optimization. Two optimized formulations were characterized using in vitro re-dispersibility and cytotoxicity on various tumor cell lines (MCF-7, CaCO2, and HepG2). The in vivo bioavailability profiles of the drug loaded in the two microemulsion systems and in the suspension form were compared. The optimized microemulsions composed of Labrafil M1944 CS (5.67%)/Tween 80 (38.71%)/Labrasol (38.71%)/water (16.92%) and Capryol 90 (0.50%)/Transcutol P (26.67%)/Tween 80 (26.67%)/Labrasol (26.67%)/water (19.50%), respectively. They revealed uniform and stable p-coumaric acid-loaded microemulsion systems with a droplet size diameter of about 10 nm. The loaded microemulsion formulations enhanced the drug re-dispersibility in contrast to the drug suspension which exhibited 5 min lag time. The loaded formulae were significantly more cytotoxic on all cell lines by 11.98–16.56 folds on MCF-7 and CaCo2 cells and 47.82–98.79 folds on HepG2 cells higher than the pure drug. The optimized microemulsions were 1.5–1.8 times more bioavailable than the drug suspension. The developed p-coumaric acid microemulsion systems could be considered a successful remedy for diverse types of cancer. [ABSTRACT FROM AUTHOR]

Cardiovascular diseases cause a large number of deaths throughout the world. No research was conducted earlier on p‐coumaric acid's effect on tachycardia, inflammation, ion pump dysfunction, and electrolyte imbalance. Hence, we appraised the above‐said parameters in isoproterenol‐induced myocardial infarcted rats. This investigation included 24 male albino Wistar rats in 4 groups. Normal control Group 1, p‐coumaric acid (8 mg/kg body weight) alone treated Group 2, Isoproterenol (100 mg/kg body weight) induced myocardial infarcted Group 3, p‐coumaric acid (8 mg/kg body weight) pretreated isoproterenol (100 mg/kg body weight) induced Group 4. After 1 day of the last dose of isoproterenol injection (day 10), rats were killed and blood and heart were taken and inflammatory markers, lipid peroxidation, nonenzymatic antioxidants, ion pumps, and electrolytes were measured. The heart rate, serum cardiac troponin‐T, serum/plasma inflammatory markers, and heart proinflammatory cytokines were raised in isoproterenol‐induced rats. Isoproterenol also enhanced plasma lipid peroxidation, lessened plasma nonenzymatic antioxidants, and altered heart ion pumps and serum and heart electrolytes. In this study, p‐coumaric acid pretreatment orally for 7 days to isoproterenol‐induced myocardial infarcted rats prevented changes in the above‐cited parameters. p‐Coumaric acid's anti‐tachycardial, anti‐inflammatory, anti‐ion pump dysfunction and anti‐electrolyte imbalance properties are the mechanisms for these cardioprotective effects. [ABSTRACT FROM AUTHOR]

Ferulic acid (FA) and p-coumaric acid (PCA) are abundantly present in commonly consumed food and beverages. Being polyphenolic compounds, they have been explored for their antioxidant and anti-inflammatory properties. Based on our previous study, we selected these two compounds to further investigate their potential in lipopolysaccharide (LPS)-induced sickness behavior and the ensuing neuroinflammation by specifically focusing on the NLRP3 inflammasome pathway. Male Swiss albino mice were divided into nine groups (n = 6) consisting of Normal Control, LPS, fluoxetine (FLX), FA40, FA160, FA640, PCA40, PCA160, and PCA640 respectively. Each group received respective FA or PCA treatment except Normal Control and LPS, which received the vehicle, carboxymethylcellulose 0.25% w/v. All groups were challenged with LPS 1.5 mg/kg, intraperitoneally except the Normal Control group, which received saline. Behavioral assessments were performed between 1–2 h, and the whole brains were collected at 3 h post-LPS administration. LPS-induced sickness behavior was characterized by significantly reduced spontaneous activity and high immobility time. The expression of NLRP3, ASC, caspase-1 and IL-1β was significantly increased, along with the levels of brain IL-1β suggesting the assembly and activation of NLRP3 inflammasome pathway. Furthermore, the major cytokines involved in sickness behavior, IL-6 and TNF-α were also significantly elevated with the accompanied lipid peroxidation. The results of this study emphasize that within the employed dose ranges of both FA and PCA, both the compounds were effective at blocking the activation of the NLRP3 inflammasome pathway and thereby reducing the release of IL-1β and the sickness behavior symptoms. There was a prominent effect on cytokine levels and lipid peroxidation as well. [ABSTRACT FROM AUTHOR]

Phenolic acids are derivatives of benzoic and cinnamic acids, which possess important biological activities at certain concentrations. Trans-cinnamic acid (t-CA) and its derivatives, such as p-coumaric acid (p-CA) and ferulic acid (FA) have been shown to have antibacterial activity against various Gram-positive and -negative bacteria. However, there is limited information available concerning the antibacterial mode of action of these phenolic acids. In this study, we aimed to ascertain metabolic alterations associated with exposure to t-CA, p-CA, and FA in Escherichia coli BW25113 using a nuclear magnetic resonance (NMR)-based metabolomics approach. The results showed that t-CA, p-CA, and FA treatments led to significant changes (p < 0.05) in the concentration of 42, 55, and 74% of the identified metabolites in E. coli, respectively. Partial least-squares discriminant analysis (PLS-DA) revealed a clear separation between control and phenolic acid groups with regard to metabolic response. Moreover, it was found that FA and p-CA treatment groups were clustered closely together but separated from the t-CA treatment group. Arginine, putrescine, cadaverine, galactose, and sucrose had the greatest impact on group differentiation. Quantitative pathway analysis demonstrated that arginine and proline, pyrimidine, glutathione, and galactose metabolisms, as well as aminoacyl-tRNA and arginine biosyntheses, were markedly affected by all phenolic acids. Finally, the H2O2 content of E. coli cells was significantly increased in response to t-CA and p-CA whereas all phenolic acids caused a dramatic increase in the number of apurinic/apyrimidinic sites. Overall, this study suggests that the metabolic response of E. coli cells to t-CA is relatively different from that to p-CA and FA. However, all phenolic acids had a certain impact on oxidative/antioxidant status, genomic stability, arginine-related pathways, and nucleic acid metabolism. [ABSTRACT FROM AUTHOR]

ABSTRACT: Rice husk (RH) is an abundant agro-industrial residue in Uruguay, being the disposal of it an environmental problem because of the pollution it generates. This work had as objective the obtention of phenolic compounds from RH by alkaline hydrolysis and their separation by means of aqueous two-phase systems (ATPS). An experimental design was implemented for the alkaline delignification process, studying the influence of temperature, solid/liquid (S/L) ratio and NaOH concentration on the hydrolysis yield (HY), total phenolic content (TPC), the antioxidant capacity (FRAP and TEAC) and the ferulic and p-coumaric acids content of the hydrolysates. The optimal conditions were 120 °C, S/L ratio 1/30 g/mL and 2 N NaOH. Subsequently, ATPS based on choline chloride and different surfactants, Tween 20 (Tw20) and Tween 80 (Tw80), were used to separate the phenolic compounds from the hydrolysate. The influence of the tie-line length (TLL), volume ratio (VR) and temperature were studied. Ferulic acid and p-coumaric acid migrated to the surfactant rich-phase in all cases. Systems containing Tw80 turned out to have better separation yields (SY), greater than 90%, for both phenolic acids. SY was not affected by use of different TLL and temperatures, although it was affected by changes on VR.

Recently, we identified the biosynthetic gene cluster of avenalumic acid (ava cluster) and revealed its entire biosynthetic pathway, resulting in the discovery of a diazotization-dependent deamination pathway. Genome database analysis revealed the presence of more than 100 ava cluster-related biosynthetic gene clusters (BGCs) in actinomycetes; however, their functions remained unclear. In this study, we focused on an ava cluster-related BGC in Kutzneria albida (cma cluster), and revealed that it is responsible for p-coumaric acid biosynthesis by heterologous expression of the cma cluster and in vitro enzyme assays using recombinant Cma proteins. The ATP-dependent diazotase CmaA6 catalyzed the diazotization of both 3-aminocoumaric acid and 3-aminoavenalumic acid using nitrous acid in vitro. In addition, the high efficiency of the CmaA6 reaction enabled us to perform a kinetic analysis of AvaA7, which confirmed that AvaA7 catalyzes the denitrification of 3-diazoavenalumic acid in avenalumic acid biosynthesis. This study deepened our understanding of the highly reducing type II polyketide synthase system as well as the diazotization-dependent deamination pathway for the production of avenalumic acid or p-coumaric acid.

Background: Gray mold, caused by Botrytis cinerea, is one of the major fungal diseases in agriculture. Biological methods are preferred over chemical fungicides to control gray mold since they are less toxic to the environment and could induce the resistance to pathogens in plants. In this work, we try to understand if ginseng defense to B. cinerea could be induced by fungal hypovirulent strain △BcSpd1. BcSpd1 encodes Zn(II)2Cys6 transcription factor which regulates fungal pathogenicity and we recently reported △BcSpd1 mutants reduced fungal virulence. Methods: We performed transcriptomic analysis of the host to investigate the induced defense response of ginseng treated by B. cinerea △BcSpd1. The metabolites in ginseng flavonoids pathway were determined by UPLC-ESI-MS/MS and the antifungal activates were then performed. Results: We found that △BcSpd1 enhanced the ginseng defense response when applied to healthy ginseng leaves and further changed the metabolism of flavonoids. Compared with untreated plants, the application of △BcSpd1 on ginseng leaves significantly increased the accumulation of p-coumaric acid and myricetin, which could inhibit the fungal growth. Conclusion: B. cinerea △BcSpd1 could effectively induce the medicinal plant defense and is referred to as the biological control agent in ginseng disease management.

Dynamic regulation has been proved efficient in controlling gene expression at transcriptional, translational, and post-translational level. However, the dynamic regulation at gene replication level has been rarely explored so far. In this study, we established dynamic regulation at gene copy level through engineering controllable plasmid replication to dynamically control the gene expression. Prototypic genetic circuits with different control logic were applied to enable diversified dynamic behaviors of gene copy. To explore the applicability of this strategy, the dynamic gene copy control was employed in regulating the biosynthesis of p-coumaric acid, which resulted in an up to 78% increase in p-coumaric acid titer to 1.69 g/L in shake flasks. These results indicated the great potential of applying dynamic gene copy control for engineering biosynthesis of valuable compounds in metabolic engineering.

Abstract Background Phenylpropanoids such as p-coumaric acid represent important precursors for the synthesis of a broad range of plant secondary metabolites including stilbenoids, flavonoids, and lignans, which are of pharmacological interest due to their health-promoting properties. Although extraction from plant material or chemical synthesis is possible, microbial synthesis of p-coumaric acid from glucose has the advantage of being less expensive and more resource efficient. In this study, Corynebacterium glutamicum was engineered for the production of the plant polyphenol precursor p-coumaric acid from glucose. Results Heterologous expression of the tyrosine ammonia-lyase encoding gene from Flavobacterium johnsoniae enabled the conversion of endogenously provided tyrosine to p-coumaric acid. Product consumption was avoided by abolishing essential reactions of the phenylpropanoid degradation pathway. Accumulation of anthranilate as a major byproduct was eliminated by reducing the activity of anthranilate synthase through targeted mutagenesis to avoid tryptophan auxotrophy. Subsequently, the carbon flux into the shikimate pathway was increased, phenylalanine biosynthesis was reduced, and phosphoenolpyruvate availability was improved to boost p-coumaric acid accumulation. A maximum titer of 661 mg/L p-coumaric acid (4 mM) in defined mineral medium was reached. Finally, the production strain was utilized in co-cultivations with a C. glutamicum strain previously engineered for the conversion of p-coumaric acid into the polyphenol resveratrol. These co-cultivations enabled the synthesis of 31.2 mg/L (0.14 mM) resveratrol from glucose without any p-coumaric acid supplementation. Conclusions The utilization of a heterologous tyrosine ammonia-lyase in combination with optimization of the shikimate pathway enabled the efficient production of p-coumaric acid with C. glutamicum. Reducing the carbon flux into the phenylalanine and tryptophan branches was the key to success along with the introduction of feedback-resistant enzyme variants.

Background: Apple cider vinegar (ACV) is most commonly used in sweet and sour Chinese cuisine. This study aimed to overcome the side effects of raw ACV, by increasing the concentration of p-Coumaric acid (p-CA), a polyphenolic component of ACV. The combination (diluted ACV with p-CA) has also been compared with individual raw ACV and p-CA to confirm if the overall dosage of the ACV can be reduced to avoid side effects and if the combination therapy had any better effect than the individual component itself. Objective: To evaluate and compare antioxidant, antimicrobial, and wound healing effects of ACV and p-CA combination with individual components ACV and p-CA. Methods: The antimicrobial properties of the samples were assessed by determining the Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using the broth microdilution method, and zone of inhibition (ZOI) and an ex ovo study was also done to evaluate the antimicrobial effect of the samples in live embryo. For the evaluation of the test samples excision wound was created in Swiss male albino mice weighing 25–30 g of either sex to estimate parameters like wound contraction rate, WBC and platelet count, SOD and GSH levels. Histopathological analysis of the skin was also done. Result: In DPPH and ABTS antioxidant assays, the combined sample (ACV + p-CA) had lower IC50. The combination therapy also showed the best antimicrobial potential against P. aeruginosa and B. subtilis. In this context, ex-ovo antimicrobial study results showed that diluted ACV + p-CA treated live embryo samples had the least bacterial growth after 48 h, in comparison with non-treatment group as well as individually ACV and p-CA treated samples. In vivo study depicted that the highest dose of the combination test sample had the best wound contraction rate and antioxidant marker enzymes elevation compared to diseased control proving the potency to restore the wound healing progression. Conclusion: ACV and p-CA combination can be used with daily intake as this combination can prevent microbial contamination and oxidative stress additionally can repair wounds more safely than raw ACV.