Shunt Pathway Research Articles

A cascade bilayer electron transport layer toward efficient and stable Ruddlesden‐Popper perovskite solar cells

Optimal interfaces play a key role in charge transport/recombination characteristics and minimized potential loss of perovskite solar cells (PSCs). Currently, n-type oxide semiconductors are the most common electron transport layer (ETL) material, but the imperfect electronic structure, unfavorable band alignment, and corresponding poor interface of ETL/perovskite absorber remain a great challenge for achieving a high photovoltaic performance of PSCs. Here, we combine a fullerene derivative or a non-fullerene organic semiconductor with tin(IV) oxide (SnO2) to promise much-improved surface and electronic structure of ETL interface in Ruddlesden-Popper perovskites (RPPs)-based photovoltaic devices. The organic semiconductors fill a bumpy surface of SnO2 to make the surface smoother, which effectively avoids the shunt pathways at the interface of the ETL/RPP absorber layer, and thereby suppresses unintended electron-hole recombination. The organic-inorganic bilayered ETLs also improve the electronic structure of the SnO2 surface, which provides favorable optoelectronic properties such as quick electron extraction and elongated carrier lifetime in the device. Moreover, the hydrophobic organic semiconductor is helpful in not only growing a uniform, compact, and largely grained RPP absorber layer but also improving the stability of the perovskite absorber layer. Benefiting from the optimized bilayer ETLs, the power conversion efficiency is obviously enhanced up to 10.17% in the RPP-based photovoltaic devices. More importantly, the bilayer ETLs allow improved long-term stability in ambient storage of the devices, providing a viable path to design practical interfaces of efficient and stable RPP-based PSCs. Highlights Hybrid bilayer electron transport layer is designed to enhance the efficiency of Ruddlesden-Popper perovskite solar cells. Phenyl-C61-butyric acid methyl ester and ITIC are inserted as n-type organic semiconductors onto SnO2 film for the bilayer electron transport layer. Tailored optoelectronic properties of the bilayer electron transport layers deliver 10.17% efficiency of Ruddlesden-Popper perovskite solar cells.

Electrochemical Screening of Contact Layers for Metal Halide Perovskites

Optimizing selective contact layers in photovoltaics is necessary to yield high performing stable devices. However, this has been difficult for perovskites due to their complex interfacial defects that affect carrier concentrations in the active layer as well as charge transfer and recombination at the interface. Using vacuum thermally-evaporated tin oxide as a case study, we highlight electrochemical tests that are simple yet screen device-relevant contact layer properties, making them useful for process development and quality control. Specifically, we show that cyclic voltammetry and potentiostatic chronoamperometry correlate to key performance parameters in completed devices as well as other material/interfacial properties relevant to devices such as shunt pathways and chemical composition. Having fast, reliable, scalable, and actionable probes of electronic properties is increasingly important as halide perovskite photovoltaics approach their theoretical limits and scale to large-area devices.

Effect of drought stress on wheat (Triticum durum) growth and metabolism: insight from GABA shunt, reactive oxygen species and dehydrin genes expression.

Activation of γ-aminobutyric acid (GABA) shunt pathway and upregulation of dehydrins are involved in metabolic homeostasis and protective mechanisms against drought stress. Seed germination percentage, seedling growth, levels of GABA, alanine, glutamate, malondialdehyde (MDA), and the expression of glutamate decarboxylase (GAD ) and dehydrin (dhn and wcor ) genes were examined in post-germination and seedlings of four durum wheat (Triticum durum L.) cultivars in response to water holding capacity levels (80%, 50%, and 20%). Data showed a significant decrease in seed germination percentage, seedling length, fresh and dry weight, and water content as water holding capacity level was decreased. Levels of GABA, alanine, glutamate, and MDA were significantly increased with a negative correlation in post-germination and seedling stages as water holding capacity level was decreased. Prolonged exposure to drought stress increased the GAD expression that activated GABA shunt pathway especially at seedlings growth stage to maintain carbon/nitrogen balance, amino acids and carbohydrates metabolism, and plant growth regulation under drought stress. The mRNA transcripts of dhn and wcor significantly increased as water availability decreased in all wheat cultivars during the post-germination stage presumably to enhance plant tolerance to drought stress by cell membrane protection, cryoprotection of enzymes, and prevention of reactive oxygen species (ROS) accumulation. This study showed that the four durum wheat cultivars responded differently to drought stress especially during the seedling growth stage which might be connected with ROS scavenging systems and the activation of antioxidant enzymes that were associated with activation of GABA shunt pathway and the production of GABA in durum seedlings.

Phenotypic effects from the expression of a deregulated AtGAD1 transgene and GABA pathway suppression mutants in maize.

Glutamate decarboxylase (GAD; EC 4.1.1.15) catalyzes the irreversible decarboxylation of glutamate to produce γ-aminobutyric acid (GABA); a ubiquitous non-protein amino acid involved in the regulation of several aspects of plant metabolism and physiology. To study the function of GAD and GABA in maize, we have; 1) introduced native and deregulated forms of AtGAD1 into maize with the intent of increasing the synthesis of GABA and 2) introduced constructs into maize designed to suppress the activity of several GABA shunt, GABA transport and GABA pathway genes. Maize plants expressing the deregulated AtGAD1 exhibit a severe chlorosis and retarded growth phenotype and have high levels of GABA, and Ca++/CaM-independent GAD activity. Plants expressing the suppression constructs for GABA biosynthetic and transport pathway genes had no observable phenotype whereas a knockout of GABA catabolic pathway genes led to growth and developmental defects under standard growth conditions. The implications of this study to our understanding of the action and function of GABA and GAD in crops are discussed.

Trichoderma reesei Contains a Biosynthetic Gene Cluster That Encodes the Antifungal Agent Ilicicolin H.

The trili biosynthetic gene cluster (BGC) from the well-studied organism Trichoderma reesei was studied by heterologous expression in the fungal host Aspergillus oryzae. Coexpression of triliA and triliB produces two new acyl tetramic acids. Addition of the ring-expanding cytochrome P450 encoded by triliC then yields a known pyridone intermediate to ilicicolin H and a new chain-truncated shunt metabolite. Finally, addition of the intramolecular Diels-Alderase encoded by triliD affords a mixture of 8-epi ilicicolin H and ilicicolin H itself, showing that the T. reesei trili BGC encodes biosynthesis of this potent antifungal agent. Unexpected A. oryzae shunt pathways are responsible for the production of the new compounds, emphasising the role of fungal hosts in catalysing diversification reactions.

Phytosulfokine \u03b1 (PSK\u03b1) delays senescence and reinforces SUMO1/SUMO E3 ligase SIZ1 signaling pathway in cut rose flowers (Rosa hybrida cv. Angelina)

Roses are widely used as cut flowers worldwide. Petal senescence confines the decorative quality of cut rose flowers, an impressively considerable economic loss. Herein, we investigated the SUMO1/SUMO E3 ligase SIZ1 signaling pathway during bud opening, and petal senescence of cut rose flowers. Our results exhibited that the higher expression of SUMO1 and SUMO E3 ligase SIZ1 during bud opening was accompanied by lower endogenous H2O2 accumulation arising from higher expression and activities of SOD, CAT, APX, and GR, promoting proline accumulation by increasing P5CS expression and activity and enhancing GABA accumulation by increasing GAD expression and activity. In harvested flowers, lower expressions of SUMO1 and SUMO E3 ligase SIZ1 during petal senescence were associated with higher endogenous H2O2 accumulation due to lower expression and activities of SOD, CAT, APX, and GR. Therefore, promoting the activity of the GABA shunt pathway as realized by higher expression and activities of GABA-T and SSADH accompanied by increasing OAT expression and activity for sufficiently supply proline in rose flowers during petal senescence might serve as an endogenous antisenescence mechanism for slowing down petals senescence by avoiding endogenous H2O2 accumulation. Following phytosulfokine α (PSKα) application, postponing petal senescence in cut rose flowers could be ascribed to higher expression of SUMO1 and SUMO E3 ligase SIZ1 accompanied by higher expression and activities of SOD, CAT, APX, and GR, higher activity of GABA shunt pathway as realized by higher expression and activities of GAD, GABA-T, and SSADH, higher expression and activities of P5CS and OAT for supplying proline and higher expression of HSP70 and HSP90. Therefore, our results highlight the potential of the PSKα as a promising antisenescence signaling peptide in the floriculture industry for postponing senescence and extending the vase life of cut rose flowers.

Contribution of pre-storage melatonin application to chilling tolerance of some mango fruit cultivars and relationship with polyamines metabolism and γ-aminobutyric acid shunt pathway

The efficacy of pre-storage application of 100 µM melatonin on conferring chilling tolerance in fruit of four mango cultivars stored at 5 ± 1 ºC for 28 d was investigated. ‘Langra’ and ‘Gulab Jamun’ mangoes had maximum and minimum chilling tolerance, respectively. Accumulation of endogenous polyamines and γ-aminobutyric acid (GABA) in the peel and the pulp of ‘Langra’ fruit treated with melatonin was a possible reason for its chilling tolerance. Accumulation of endogenous polyamines was associated with higher arginine decarboxylase (ADC) and ornithine decarboxylase (ODC), and lower diamine oxidase (DAO) and polyamine oxidase (PAO) activities in the peel and pulp. Additionally, higher GABA shunt (Glutamate decarboxylase and GABA-transaminase) activity was associated with elevated GABA content in ‘Langra’ mangoes. The limited effectiveness of melatonin treatment for chilling injury alleviation in ‘Chaunsa’ was probably due to higher polyamines accumulation accompanied with lower PAO and DAO activity in the peel and pulp of the fruit, in addition to higher ADC and ODC in the peel.

In This Issue

In This Issue

Multiple strategies for metabolic engineering of Escherichia coli for efficient production of glycolate.

Glycolate is a bulk chemical with wide applications in the textile, food processing, and pharmaceutical industries. Glycolate can be produced from glucose via the glycolysis and glyoxylate shunt pathways, followed by reduction to glycolate. However, two problems limit the productivity and yield of glycolate when using glucose as the sole carbon source. The first is a cofactor imbalance in the production of glycolate from glucose via the glycolysis pathway, since NADPH is required for glycolate production, while glycolysis generates NADH. To rectify this imbalance, the NADP+ -dependent glyceraldehyde 3-phosphate dehydrogenase GapC from Clostridium acetobutylicum was introduced to generate NADPH instead of NADH in the oxidation of glyceraldehyde 3-phosphate during glycolysis. The soluble transhydrogenase SthA was further eliminated to conserve NADPH by blocking its conversion into NADH. The second problem is an unfavorable carbon flux distribution between the tricarboxylic acid cycle and the glyoxylate shunt. To solve this problem, isocitrate dehydrogenase (ICDH) was eliminated to increase the carbon flux of glyoxylate and thereby improve the glycolate titer. After engineering through the integration of gapC, combined with the inactivation of ICDH, SthA, and by-product pathways, as well as the upregulation of the two key enzymes isocitrate lyase (encoding by aceA), and glyoxylate reductase (encoding by ycdW), the glycolate titer increased to 5.3 g/L with a yield of 1.89 mol/mol glucose. Moreover, an optimized fed-batch fermentation reached a titer of 41 g/L with a yield of 1.87 mol/mol glucose after 60 h.

Temperature and solvent exposure response of three fatty acid peroxygenase enzymes for application in industrial enzyme processes

Free fatty acids (FFAs) are a useful feedstock for a range of industrial chemical synthesis applications. However, efficiently converting FFAs to molecules for biofuel and other high-value chemicals requires more efficient and cost-effective catalysts. Cytochrome P450 fatty acid peroxygenases (CYP152) have a unique chemistry that allows use of the peroxide shunt pathway for biochemical conversion of FFAs. Known CYP152s are heat labile, however, requiring characterization of more thermotolerant versions for use in industrial applications. A fatty acid peroxygenase from Bacillus methanolicus (CYP152K6) was shown here to have a higher optimal reaction temperature than OleT (CYP152L1). CYP152K6 was stable up to 50 °C and showed great stability in 3% acetone and dimethylformamide. Stability in solvents helps the enzyme's substrates remain soluble in solution for more efficient catalysis, and heat stability allows enzymes to remain active longer during industrial processes.

One stone two birds: Biosynthesis of 3-hydroxypropionic acid from CO2 and syngas-derived acetic acid in Escherichia coli

Syngas, which contains large amount of CO2 as well as H2 and CO, can be convert to acetic acid chemically or biologically. Nowadays, acetic acid become a cost-effective nonfood-based carbon source for value-added biochemical production. In this study, acetic acid and CO2 were used as substrates for the biosynthesis of 3-hydroxypropionic acid (3-HP) in metabolically engineered Escherichia coli carrying heterogeneous acetyl-CoA carboxylase (Acc) from Corynebacterium glutamicum and codon-optimized malonyl-CoA reductase (MCR) from Chloroflexus aurantiacus. Strategies of metabolic engineering included promoting glyoxylate shunt pathway, inhibiting fatty acid synthesis, dynamic regulating of TCA cycle, and enhancing the assimilation of acetic acid. The engineered strain LNY07(M*DA) accumulated 15.8 g/L of 3-HP with the yield of 0.71 g/g in 48 h by whole-cell biocatalysis. Then, syngas-derived acetic acid was used as substrate instead of pure acetic acid. The concentration of 3-HP reached 11.2 g/L with the yield of 0.55 g/g in LNY07(M*DA). The results could potentially contribute to the future development of an industrial bioprocess of 3-HP production from syngas-derived acetic acid.

Neuroprotective Function of High Glycolytic Activity in Astrocytes: Common Roles in Stroke and Neurodegenerative Diseases.

Astrocytes (also, astroglia) consume huge amounts of glucose and produce lactate regardless of sufficient oxygen availability, indicating a high capacity for aerobic glycolysis. Glycolysis in astrocytes is activated in accordance with neuronal excitation and leads to increases in the release of lactate from astrocytes. Although the fate of this lactate remains somewhat controversial, it is believed to fuel neurons as an energy substrate. Besides providing lactate, astrocytic glycolysis plays an important role in neuroprotection. Among the minor pathways of glucose metabolism, glucose flux to the pentose-phosphate pathway (PPP), a major shunt pathway of glycolysis, is attracting research interest. In fact, PPP activity in astrocytes is five to seven times higher than that in neurons. The astrocytic PPP plays a key role in protecting neurons against oxidative stress by providing neurons with a reduced form of glutathione, which is necessary to eliminate reactive oxygen species. Therefore, enhancing astrocytic glycolysis might promote neuronal protection during acute ischemic stroke. Contrariwise, the dysfunction of astrocytic glycolysis and the PPP have been implicated in the pathogenesis of various neurodegenerative diseases such as Parkinson’s disease, Alzheimer’s disease, and amyotrophic lateral sclerosis, since mitochondrial dysfunction and oxidative stress trigger and accelerate disease progression.

Physiological and Biochemical Characterization of the GABA Shunt Pathway in Pea (Pisum sativum L.) Seedlings under Drought Stress

The physiological and biochemical role of the γ-aminobutyric acid (GABA) shunt pathway in green pea seedlings (Pisum sativum L.) was studied in response to soil water holding capacity levels: 80%, 60%, 40%, 20%, and 10% grown under continuous light at 25 °C for 7 days and 14 days, separately. Characterization of seeds germination pattern, seedlings growth (plant height, fresh and dry weight, and chlorophyll contents), GABA shunt metabolite (GABA, glutamate, and alanine) levels, total protein and carbohydrate levels, and oxidative damage (MDA level) were examined. Data showed a significant effect of drought stress on seed germination, plant growth, GABA shunt metabolites level, total protein and carbohydrate contents, and MDA level. A significant decline in seed germination percentage was recorded at a 20% drought level, which indicated that 20% of soil water holding capacity is the threshold value of water availability for normal germination after 14 days. Seedling fresh weight, dry weight, and plant height were significantly reduced with a positive correlation as water availability was decreased. There was a significant decrease with a positive correlation in Chl a and Chl b contents in response to 7 days and 14 days of drought. GABA shunt metabolites were significantly increased with a negative correlation as water availability decreased. Pea seedlings showed a significant increase in protein content as drought stress was increased. Total carbohydrate levels increased significantly when the amount of water availability decreased. MDA content increased slightly but significantly after 7 days and sharply after 14 days under all water stress levels. The maximum increase in MDA content was observed at 20% and 10% water levels. Overall, the significant increases in GABA, protein and carbohydrate contents were to cope with the physiological impact of drought stress on Pisum sativum L. seedlings by maintaining cellular osmotic adjustment, protecting plants from oxidative stress, balancing carbon and nitrogen (C:N) metabolism, and maintaining cell metabolic homeostasis and cell turgor. The results presented in this study indicated that severe (less than 40% water content of the holding capacity) and long-term drought stress should be avoided during the germination stage to ensure proper seedling growth and metabolism in Pisum sativum L.

Hypoxia-induced increase in GABA content is essential for restoration of membrane potential and preventing ROS-induced disturbance to ion homeostasis

When plants are exposed to hypoxic conditions, the level of γ-aminobutyric acid (GABA) in plant tissues increases by several orders of magnitude. The physiological rationale behind this elevation remains largely unanswered. By combining genetic and electrophysiological approach, in this work we show that hypoxia-induced increase in GABA content is essential for restoration of membrane potential and preventing ROS-induced disturbance to cytosolic K+ homeostasis and Ca2+ signaling. We show that reduced O2 availability affects H+-ATPase pumping activity, leading to membrane depolarization and K+ loss via outward-rectifying GORK channels. Hypoxia stress also results in H2O2 accumulation in the cell that activates ROS-inducible Ca2+ uptake channels and triggers self-amplifying “ROS-Ca hub,” further exacerbating K+ loss via non-selective cation channels that results in the loss of the cell's viability. Hypoxia-induced elevation in the GABA level may restore membrane potential by pH-dependent regulation of H+-ATPase and/or by generating more energy through the activation of the GABA shunt pathway and TCA cycle. Elevated GABA can also provide better control of the ROS-Ca2+ hub by transcriptional control of RBOH genes thus preventing over-excessive H2O2 accumulation. Finally, GABA can operate as a ligand directly controlling the open probability and conductance of K+ efflux GORK channels, thus enabling plants adaptation to hypoxic conditions.

GABA shunt: a key-player in mitigation of ROS during stress

Unfavorable environmental conditions such as heat, cold, drought, metal/metalloid toxicity, and pathogens enhance production of intra-and inter-cellular levels of reactive oxygen species (ROS) in plants. ROS, acting as signaling molecules, activate signal transduction pathways in response to various stresses. Alternatively, ROS cause irreversible cellular damage due to lipid peroxidation, oxidation of protein, inactivation of enzymes, DNA damage, and interact with other vital constituents of plant cells through their strong oxidative properties, which drastically alter plant morphological structures, becoming disadvantageous for survival and productivity. Higher plants have complex defense systems to scavenge ROS. Being a central molecule of the defense system, gamma-aminobutyric acid (GABA) is ubiquitous from prokaryotes to eukaryotes cells. GABA helps mitigate ROS in plants and GABA shunt pathway plays a key role either as metabolites or endogenous signaling molecules in several regulatory mechanisms under stress conditions. The GABA transporters (GATs) being activated with the attachment of GABA under environmental stress stimuli facilitate high content of Ca2+ into the cytosol. Ca2+ combines with calmodulin (CaM) -binding domain that activates the glutamate decarboxylase (GAD) enzyme for the conversion of glutamate into GABA. This synchronized process regulates GABA shunt gene expressions under stress conditions and improves defense mechanisms in plants. This review highlights the regulatory aspects of GABA shunt pathway for ROS production as well as in the defense mechanism of plants.

Solanum lycopersicum Seedlings. Metabolic Responses Induced by the Alkamide Affinin.

Alkamides have been observed to interact in different ways in several superior organisms and have been used in traditional medicine in many countries e.g., to relieve pain. Previous studies showed that affinin when applied to other plant species induces prominent changes in the root architecture and induces transcriptional adjustments; however, little is known about the metabolic pathways recruited by plants in response to alkamides. Previous published work with Arabidopsis seedlings treated in vitro with affinin at 50 µM significantly reduced primary root length. In tomato seedlings, that concentration did not reduce root growth but increase the number and length of lateral roots. Non-targeted metabolomic analysis by Gas Chromatography couplet to Mass Spectrometry (GC/EIMS) showed that, in tomato seedlings, affinin increased the accumulation of several metabolites leading to an enrichment of several metabolic pathways. Affinin at 100 µM alters the accumulation of metabolites such as organic acids, amino acids, sugars, and fatty acids. Finally, our results showed a response possibly associated with nitrogen, GABA shunt and serine pathways, in addition to a possible alteration in the mitochondrial electron transport chain (ETC), interesting topics to understand the molecular and metabolic mechanisms in response to alkamide in plants.

Modulation of lanosterol synthase drives 24,25-epoxysterol synthesis and oligodendrocyte formation

Small molecules that promote the formation of new myelinating oligodendrocytes from oligodendrocyte progenitor cells (OPCs) are potential therapeutics for demyelinating diseases. We recently established inhibition of specific cholesterol biosynthesis enzymes and resulting accumulation of 8,9-unsaturated sterols as a unifying mechanism through which many such molecules act. To identify more potent sterol enhancers of oligodendrocyte formation, we synthesized a collection of 8,9-unsaturated sterol derivatives and found that 24,25-epoxylanosterol potently promoted oligodendrocyte formation. In OPCs, 24,25-epoxylanosterol was metabolized to 24,25-epoxycholesterol via the epoxycholesterol shunt pathway. Increasing flux through the epoxycholesterol shunt using genetic manipulation or small-molecule inhibition of lanosterol synthase (LSS) increased endogenous 24,25-epoxycholesterol levels and OPC differentiation. Notably, exogenously supplied 24,25-epoxycholesterol promoted oligodendrocyte formation despite lacking an 8,9-unsaturation. This work highlights epoxycholesterol shunt usage, controlled by inhibitors of LSS, as a target to promote oligodendrocyte formation. Additionally, sterols beyond the 8,9-unsaturated sterols, including 24,25-epoxycholesterol, drive oligodendrocyte formation.

Effect of Caralluma tuberculata on regulation of carbohydrate metabolizing genes in alloxan-induced rats

Ethnopharmacological relevanceCaralluma tuberculata (C. tuberculata) has traditionally been used in Pakistan and other parts of the world as a folk treatment for diabetes mellitus. A few studies indicated its antihyperglycemic effect, however, the mystery remained unfolded as how did it modify the pathophysiological condition. Aim of studyHence, this study aimed to explore underlying mechanism(s) for its hypoglycemic activity at biochemical and molecular levels. Materials and methodsMethanol extract (ME) of C. tuberculata as well as its hexane (HF) and aqueous (AF) fractions were explored for their effect on total glycogen in liver and skeletal muscle of alloxan-induced rats by spectroscopy. Moreover, the expression of genes related to hepatic carbohydrate metabolizing enzymes was quantified. At molecular level, mRNA expression of glucose transporter 2 (GLUT-2), glycogen synthase (GS), glucokinase (GK), hexokinase 1 (HK-1), pyruvate kinase (PK), glucose 6 phosphate dehydrogenase (G-6-PDH), pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK) and glucose 6 phosphatase (G-6-Pase) was determined by using quantitative real time polymerase chain reaction (qRT-PCR) after administration of ME (350 mg), HF(3 mg), AF (10 mg) and metformin (500 mg). The doses were administered twice daily according to per kg of body weight. ResultsA significant reduction in hepatic and skeletal muscle glycogen content was exhibited. The data of qRT-PCR revealed that gene's expression of GLUT-2 was significantly decreased after treatment with ME and HF, whilst it was unaltered by AF, however, a significant decrease was observed in genes corresponding to GS, GK and HK-1 after treatment with ME. Similarly, there was a significant decrease in expression of genes corresponding to GS, GK and HK-1 following treatment with HF. Surprisingly, post-treatment with AF didn't modify the gene's expression of GS and GK, whilst it caused a profound decrease in expression of HK-1 gene. Contrarily, the expression of gene related to PK was significantly up-regulated post-administration with ME, HF and AF. The expression levels of G-6-PDH, however, remained unaltered after treatment with the experimental extract and fractions of the plant. In addition, HF and AF did not cause any modification in PEPCK, whereas ME caused a significant down-regulation of the gene. Treatment with all the extract and fractions of the plant caused a substantial decrease in the gene's expression of PC, while there was a significant increase in the expression of gene related to G-6-Pase. ConclusionThe three experimental extract and fractions caused a substantial decrease in glycogen content in liver and skeletal muscle tissues. The analysis by qRT-PCR showed that glucose transport via GLUT-2 was profoundly declined by ME and HF. The expression of genes related to various metabolic pathways involved in metabolism of carbohydrate in hepatocytes revealed explicitly that the ME, HF and AF decreased the phenomena of glycogenesis and gluconeogenesis. Contrarily, all the extract and fractions of the plant activated glycogenolysis and glycolysis but did not modify the pentose phosphate shunt pathway.

Enhanced glycolic acid yield through xylose and cellobiose utilization by metabolically engineered Escherichia coli.

Microbial biorefinery is a promising route toward sustainable production of glycolic acid (GA), a valuable raw material for various industries. However, inherent microbial GA production has limited substrate consumption using either D-xylose or D-glucose as carbon catabolite repression (CCR) averts their co-utilization. To bypass CCR, a GA-producing strain using D-xylose via Dahms pathway was engineered to allow cellobiose uptake. Unlike glucose, cellobiose was assimilated and intracellularly degraded without repressing D-xylose uptake. The final GA-producing E. coli strain (CLGA8) has an overexpressed cellobiose phosphorylase (cep94A) from Saccharophagus degradans 2-40 and an activated glyoxylate shunt pathway. Expression of cep94A improved GA production reaching the maximum theoretical yield (0.51gGAg-1 xylose), whereas activation of glyoxylate shunt pathway enabled GA production from cellobiose, which further increased the GA titer(2.25 g GA L-1). To date, this is the highest reported GA yield from D-xylose through Dahms pathway in an engineered E. coli with cellobiose as co-substrate.

Non-invasive skin sampling of tryptophan/kynurenine ratio in vitro towards a skin cancer biomarker

The tryptophan to kynurenine ratio (Trp/Kyn) has been proposed as a cancer biomarker. Non-invasive topical sampling of Trp/Kyn can therefore serve as a promising concept for skin cancer diagnostics. By performing in vitro pig skin permeability studies, we conclude that non-invasive topical sampling of Trp and Kyn is feasible. We explore the influence of different experimental conditions, which are relevant for the clinical in vivo setting, such as pH variations, sampling time, and microbial degradation of Trp and Kyn. The permeabilities of Trp and Kyn are overall similar. However, the permeated Trp/Kyn ratio is generally higher than unity due to endogenous Trp, which should be taken into account to obtain a non-biased Trp/Kyn ratio accurately reflecting systemic concentrations. Additionally, prolonged sampling time is associated with bacterial Trp and Kyn degradation and should be considered in a clinical setting. Finally, the experimental results are supported by the four permeation pathways model, predicting that the hydrophilic Trp and Kyn molecules mainly permeate through lipid defects (i.e., the porous pathway). However, the hydrophobic indole ring of Trp is suggested to result in a small but noticeable relative increase of Trp diffusion via pathways across the SC lipid lamellae, while the shunt pathway is proposed to slightly favor permeation of Kyn relative to Trp.