•Escherichia coli was engineered for the production of caffeic acid.•Tyrosine ammonia lyase (TAL) converted 3mM of tyrosine to 2.62mM p-coumaric acid.•TAL and 4-coumarate 3-hydroxylase (C3H) ...converted tyrosine in 1mM caffeic acid.•This is the first study that shows caffeic acid production using TAL and CYP199A2.•TAL and CYP199A2 converted tyrosine in 1.56mM caffeic acid.
Caffeic acid is a plant secondary metabolite and its biological synthesis has attracted increased attention due to its beneficial effects on human health. In this study, Escherichia coli was engineered for the production of caffeic acid using tyrosine as the initial precursor of the pathway. The pathway design included tyrosine ammonia lyase (TAL) from Rhodotorula glutinis to convert tyrosine to p-coumaric acid and 4-coumarate 3-hydroxylase (C3H) from Saccharothrix espanaensis or cytochrome P450 CYP199A2 from Rhodopseudomonas palustris to convert p-coumaric acid to caffeic acid. The genes were codon-optimized and different combinations of plasmids were used to improve the titer of caffeic acid. TAL was able to efficiently convert 3mM of tyrosine to p-coumaric acid with the highest production obtained being 2.62mM (472mg/L). CYP199A2 exhibited higher catalytic activity towards p-coumaric acid than C3H. The highest caffeic acid production obtained using TAL and CYP199A2 and TAL and C3H was 1.56mM (280mg/L) and 1mM (180mg/L), respectively. This is the first study that shows caffeic acid production using CYP199A2 and tyrosine as the initial precursor. This study suggests the possibility of further producing more complex plant secondary metabolites like flavonoids and curcuminoids.
Heterologous production of curcuminoids Rodrigues, J L; Prather, K L J; Kluskens, L D ...
Microbiology and molecular biology reviews,
03/2015, Letnik:
79, Številka:
1
Journal Article
Recenzirano
Odprti dostop
Curcuminoids, components of the rhizome of turmeric, show several beneficial biological activities, including anticarcinogenic, antioxidant, anti-inflammatory, and antitumor activities. Despite their ...numerous pharmaceutically important properties, the low natural abundance of curcuminoids represents a major drawback for their use as therapeutic agents. Therefore, they represent attractive targets for heterologous production and metabolic engineering. The understanding of biosynthesis of curcuminoids in turmeric made remarkable advances in the last decade, and as a result, several efforts to produce them in heterologous organisms have been reported. The artificial biosynthetic pathway (e.g., in Escherichia coli) can start with the supplementation of the amino acid tyrosine or phenylalanine or of carboxylic acids and lead to the production of several natural curcuminoids. Unnatural carboxylic acids can also be supplemented as precursors and lead to the production of unnatural compounds with possibly novel therapeutic properties. In this paper, we review the natural conversion of curcuminoids in turmeric and their production by E. coli using an artificial biosynthetic pathway. We also explore the potential of other enzymes discovered recently or already used in other similar biosynthetic pathways, such as flavonoids and stilbenoids, to increase curcuminoid yield and activity.
The potential use of nanostructured dipeptide self-assemblies in materials science for energy harvesting devices is a highly sought-after area of research. Specifically, aromatic cyclo-dipeptides ...containing tryptophan have garnered attention due to their wide-bandgap semiconductor properties, high mechanical rigidity, photoluminescence, and nonlinear optical behavior. In this study, we present the development of a hybrid system comprising biopolymer electrospun fibers incorporated with the chiral cyclo-dipeptide L-Tryptophan-L-Tyrosine. The resulting nanofibers are wide-bandgap semiconductors (bandgap energy 4.0 eV) consisting of self-assembled nanotubes embedded within a polymer matrix, exhibiting intense blue photoluminescence. Moreover, the cyclo-dipeptide L-Tryptophan-L-Tyrosine incorporated into polycaprolactone nanofibers displays a strong effective second harmonic generation signal of 0.36 pm/V and shows notable piezoelectric properties with a high effective coefficient of 22 pCN-1, a piezoelectric voltage coefficient of geff=1.2 VmN-1 and a peak power density delivered by the nanofiber mat of 0.16μWcm-2. These hybrid systems hold great promise for applications in the field of nanoenergy harvesting and nanophotonics.
Aim
The aim of this study was to compare the effectiveness of 2% articaine and 2% lignocaine in achieving adequate anesthesia in children between the age group of 6–13 years using inferior alveolar ...nerve block.
Methods
A triple blinded randomized controlled trial was conducted in 180 participants (90 patients- 2% articaine, 90 patients-2% lignocaine). Effectiveness of the anesthetic agent was determined at 3 points determined by subjective evaluation of pain using pain scales (FPS-R). Paired sample t-test and chi square test were performed for statistical significance.
Result
Anesthetic success for 2% articaine were 64.4%, 42.2% and 81.8% respectively. The anesthetic success of 2% lignocaine was 66.7%, 48.9% and 85.7% at point one, point two and point three respectively (
p
> 0.05).
Conclusion
This study concludes that 2% articaine in 1:2,00,000 did not demonstrate superior clinical effectiveness in comparison to 2% lignocaine.
Clinical significance
Lignocaine has always been considered the gold standard. With its unique chemical structure and increased potency, Articaine has been gaining popularity. Its efficacy in 2% concentration had not been compared to 2% lignocaine. 2% articaine did not show clinical superiority but its comparable effectiveness with lignocaine can encourage further research in using articaine in reduced concentrations to improve effectiveness.
A polymorph of glycyl-L-alanine HI.H
O is synthesized from chiral cyclo-glycyl-L-alanine dipeptide. The dipeptide is known to show molecular flexibility in different environments, which leads to ...polymorphism. The crystal structure of the glycyl-L-alanine HI.H
O polymorph is determined at room temperature and indicates that the space group is polar (P2
), with two molecules per unit cell and unit cell parameters a = 7.747 Å, b = 6.435 Å, c = 10.941 Å, α = 90°, β = 107.53(3)°, γ = 90° and V = 520.1(7) Å
. Crystallization in the polar point group 2, with one polar axis parallel to the b axis, allows pyroelectricity and optical second harmonic generation. Thermal melting of the glycyl-L-alanine HI.H
O polymorph starts at 533 K, close to the melting temperature reported for cyclo-glycyl-L-alanine (531 K) and 32 K lower than that reported for linear glycyl-L-alanine dipeptide (563 K), suggesting that although the dipeptide, when crystallized in the polymorphic form, is not anymore in its cyclic form, it keeps a memory of its initial closed chain and therefore shows a thermal memory effect. Here, we report a pyroelectric coefficient as high as 45 µC/m
K occurring at 345 K, one order of magnitude smaller than that of semi-organic ferroelectric triglycine sulphate (TGS) crystal. Moreover, the glycyl-L-alanine HI.H
O polymorph displays a nonlinear optical effective coefficient of 0.14 pm/V, around 14 times smaller than the value from a phase-matched inorganic barium borate (BBO) single crystal. The new polymorph displays an effective piezoelectric coefficient equal to deff=280 pCN-1, when embedded into electrospun polymer fibers, indicating its suitability as an active system for energy harvesting.
Peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) transcriptional coactivators are key regulators of energy metabolism-related genes and are expressed in energy-demanding ...tissues. There are several PGC-1α variants with different biological functions in different tissues. The brain is one of the tissues where the role of PGC-1α isoforms remains less explored. Here, we used a toxin-based mouse model of Parkinson's disease (PD) and observed that the expression levels of variants PGC-1α2 and PGC-1α3 in the nigrostriatal pathway increases at the onset of dopaminergic cell degeneration. This increase occurs concomitant with an increase in glial fibrillary acidic protein levels. Since PGC-1α coactivators regulate cellular adaptive responses, we hypothesized that they could be involved in the modulation of astrogliosis induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Therefore, we analysed the transcriptome of astrocytes transduced with expression vectors encoding PGC-1α1 to 1α4 by massively parallel sequencing (RNA-seq) and identified the main cellular pathways controlled by these isoforms. Interestingly, in reactive astrocytes the inflammatory and antioxidant responses, adhesion, migration, and viability were altered by PGC-1α2 and PGC-1α3, showing that sustained expression of these isoforms induces astrocyte dysfunction and degeneration. This work highlights PGC-1α isoforms as modulators of astrocyte reactivity and as potential therapeutic targets for the treatment of PD and other neurodegenerative disorders.
•PGC-1α2 and α3 expression increases in the nigrostriatal pathway of mice exposed to the MPTP model of Parkinson's disease.•PGC-1α variants control the expression of different gene sets in astrocytes in culture.•Expressing PGC-1α2 and α3 in reactive astrocytes changes inflammatory and antioxidant responses, cell motility, and viability.
Display omitted
•Alginate and extrusion technique are still widely used to encapsulate probiotics.•Gums/mucilages as encapsulating material improves the survival of encapsulated cells.•Cell ...encapsulation improves the survival of probiotics in food matrices.•Encapsulation turns non-dairy food products into alternative to deliver probiotics.
The administration of probiotic microorganisms in adequate amounts is constantly related to health benefits. To promote beneficial effects, these microorganisms must not be affected by exposure to environmental factors and must be able to adhere and colonize the human gastrointestinal tract. Several encapsulation techniques and encapsulating materials are available to produce probiotic particles, however, it is essential that the process must not be aggressive, reducing or preventing injuries and cell losses, besides, the particle properties obtained must be adequate for the proposed purpose. At the same time, the global market for supplements and probiotic foods has been growing significantly, and cell encapsulation appears as an alternative to incorporate probiotics into different food matrices. This review discusses and updates the main techniques, and the traditional and emerging polysaccharides for encapsulation of probiotic cells, as well as the advantages and possibilities of incorporating produced particles into food matrices. Currently, various scientific studies report the use of different encapsulation techniques, such as extrusion, emulsion, spray drying, spray chilling and fluidized bed to encapsulate probiotics properly. The alginate is still widely used to produce probiotic particles, however, there has been a growing interest in its total or partial substitution with others polysaccharides, such as gums, mucilages, prebiotic compounds and microbial exopolysaccharides, which improve the protection and survival of encapsulated cells and allow their incorporation into dairy and non-dairy food products.
The updated definition of prebiotic expands the range of potential applications in which emerging xylooligosaccharides (XOS) can be used. It has been demonstrated that XOS exhibit prebiotic effects ...at lower amounts compared to others, making them competitively priced prebiotics. As a result, the industry is focused on developing alternative approaches to improve processes efficiency that can meet the increasing demand while reducing costs. Recent advances have been made towards greener and more efficient processes, by applying process integration strategies to produce XOS from costless lignocellulosic residues and using genetic engineering to create microorganisms that convert these residues to XOS. In addition, collecting more in vivo data on their performance will be key to achieve regulatory claims, greatly increasing XOS commercial value.
Variability in snake venom composition is well-documented and crucial for understanding snake ecology and predicting snakebites. In this study, we characterize the venom composition and biological ...activities of newborn female and male Bothrops moojeni and their mother. Our results reveal significant differences between the venom of newborn females and males, demonstrating a broad and diverse range of proteins. The venoms of newborn females showed higher serine protease effects, increased hemorrhagic activity, and greater lethality compared to the venom of newborn males. However, no differences were observed in phospholipase A2 and coagulant activity. The differences in protein composition and toxic activities between maternal and neonatal venom, as well as between the venoms of newborn females and males, contribute to understanding the diverse outcomes of snakebites. These results underscore the importance of considering sex and ontogeny in understanding venom composition in snakes.
Curcuminoids are phenylpropanoids with high pharmaceutical potential. Herein, we report an engineered artificial pathway in Escherichia coli to produce natural curcuminoids through caffeic acid. ...Arabidopsis thaliana 4‐coumaroyl‐CoA ligase and Curcuma longa diketide‐CoA synthase (DCS) and curcumin synthase (CURS1) were used to produce curcuminoids and 70 mg/L of curcumin was obtained from ferulic acid. Bisdemethoxycurcumin and demethoxycurcumin were also produced, but in lower concentrations, by feeding p‐coumaric acid or a mixture of p‐coumaric acid and ferulic acid, respectively. Additionally, curcuminoids were produced from tyrosine through the caffeic acid pathway. To produce caffeic acid, tyrosine ammonia lyase from Rhodotorula glutinis and 4‐coumarate 3‐hydroxylase from Saccharothrix espanaensis were used. Caffeoyl‐CoA 3‐O‐methyltransferase from Medicago sativa was used to convert caffeoyl‐CoA to feruloyl‐CoA. Using caffeic acid, p‐coumaric acid or tyrosine as a substrate, 3.9, 0.3, and 0.2 mg/L of curcumin were produced, respectively. This is the first time DCS and CURS1 were used in vivo to produce curcuminoids and that curcumin was produced by feeding tyrosine. We have shown that curcumin can be produced using a pathway involvoing caffeic acid. This alternative pathway represents a step forward in the heterologous production of curcumin using E. coli.
Curcuminoids are phenylpropanoids from the plant Curcuma longa Linn, with high pharmaceutical potential. This study reports an alternative pathway for the production of curcuminoids using Escherichia coli, in which curcuminoids are produced from tyrosine through the caffeic acid pathway. The presented work represents a significant advance in the heterologous production of curcumin by E. coli.
PDF
