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Materials (Basel, Switzerland) Aug 2023Lignocellulosic materials are usually processed toward C5 and C6 corresponding sugars. Trifluoroacetic acid (TFA) is a pretreatment method to solubilize hemicellulose to...
Lignocellulosic materials are usually processed toward C5 and C6 corresponding sugars. Trifluoroacetic acid (TFA) is a pretreatment method to solubilize hemicellulose to sugars such xylose without degrading cellulose. However, this pretreatment has not been compared to other processes. Thus, this paper focuses on the techno-economic comparison of the C5-C6 production of C5-C6 as raw materials platforms using non-centrifuged sugarcane bagasse (NCSB) and wood chips (PP). Hydrolysates using TFA 2.5 M as an acid were characterized through HPLC regarding arabinose, galactose glucose, xylose, and mannose sugars. Then, simulations of the processes according to the experimental results were done. The economic assessment was performed, and compared with some common pretreatments. The mass and energy balances of the simulations indicate that the process can be compared with other pretreatments. From the economic perspective, the main operating expenditures (OpEx) are related to raw materials and capital depreciation due to the cost of TFA corrosion issues. The processes showed a CapEx and OpEx of 0.99 MUSD and 6.59 M-USD/year for NCSB, and 0.97 MUSD and 4.37 MUSD/year for PP, considering a small-scale base (1 ton/h). TFA pretreatment is innovative and promising from a techno-economic perspective.
PubMed: 37570205
DOI: 10.3390/ma16155502 -
Polymers Dec 2023This paper focuses on the changes in chemical structure and fiber morphological properties of spruce wood during 15 months of its storage in an open forest woodshed....
This paper focuses on the changes in chemical structure and fiber morphological properties of spruce wood during 15 months of its storage in an open forest woodshed. From the chemical composition, the extractives, cellulose, holocellulose, and lignin content were determined. The pH value was measured on the wood surface using a contact electrode. Acetic and formic acid, saccharides (glucose, xylose, galactose, arabinose and mannose), and polymerization degree (PD) of cellulose were analyzed using the HPLC method. Fiber length and width were determined using a fiber tester analyzer. After 15 months of storage the content of both cellulose (determined by the Seifert method) and lignin did not change; the quantity of hemicelluloses decreased by 13.2%, due to its easier degradation and less stability compared to cellulose; and the pH value dropped by one degree. HPLC analyses showed a total decrease in the cellulose DP of 9.2% and in saccharides of 40.2%, while the largest decreases were recorded in the quantity of arabinose, by 72%, in the quantity of galactose, by 61%, and in the quantity of xylose, by 43%. Organic acids were not detected due to their high volatility during wood storage. The total decrease in average fiber length was 38.2% and in width was 4.8%. An increase in the proportion of shorter fibers, and a decrease in the proportion of longer fibers, was recorded. It can be concluded that fundamental changes occurred in the wood, which could affect the quality of further products (e.g., chips, pulp, paper, particleboards).
PubMed: 38139985
DOI: 10.3390/polym15244734 -
FEMS Microbiology Ecology Apr 2024Arabinose and galactose are major, rapidly metabolized components of marine particulate and dissolved organic matter. In this study, we observed for the first time large...
Arabinose and galactose are major, rapidly metabolized components of marine particulate and dissolved organic matter. In this study, we observed for the first time large microbiomes for the degradation of arabinogalactan and report a detailed investigation of arabinogalactan utilization by the flavobacterium Maribacter sp. MAR_2009_72. Cellular extracts hydrolysed arabinogalactan in vitro. Comparative proteomic analyses of cells grown on arabinogalactan, arabinose, galactose, and glucose revealed the expression of specific proteins in the presence of arabinogalactan, mainly glycoside hydrolases (GH). Extracellular glycan hydrolysis involved five alpha-l-arabinofuranosidases affiliating with glycoside hydrolase families 43 and 51, four unsaturated rhamnogalacturonylhydrolases (GH105) and a protein with a glycoside hydrolase family-like domain. We detected expression of three induced TonB-dependent SusC/D transporter systems, one SusC, and nine glycoside hydrolases with a predicted periplasmatic location. These are affiliated with the families GH3, GH10, GH29, GH31, GH67, GH78, and GH115. The genes are located outside of and within canonical polysaccharide utilization loci classified as specific for arabinogalactan, for galactose-containing glycans, and for arabinose-containing glycans. The breadth of enzymatic functions expressed in Maribacter sp. MAR_2009_72 as response to arabinogalactan from the terrestrial plant larch suggests that Flavobacteriia are main catalysts of the rapid turnover of arabinogalactans in the marine environment.
PubMed: 38569650
DOI: 10.1093/femsec/fiae045 -
Plant Molecular Biology Dec 2023Plant cell wall-derived oligosaccharides, i.e., damage-associated molecular patterns (DAMPs), could be generated after pathogen attack or during normal plant...
Plant cell wall-derived oligosaccharides, i.e., damage-associated molecular patterns (DAMPs), could be generated after pathogen attack or during normal plant development, perceived by cell wall receptors, and can alter immunity and cell wall composition. Therefore, we hypothesised that xylo-oligosaccharides (XOS) could act as an elicitor and trigger immune responses. To test this, we treated Arabidopsis with xylobiose (XB) and investigated different parameters. XB-treatment significantly triggered the generation of reactive oxygen species (ROS), activated MAPK protein phosphorylation, and induced callose deposition. The combination of XB (DAMP) and flg22 a microbe-associated molecular pattern (MAMP) further enhanced ROS response and gene expression of PTI marker genes. RNA sequencing analysis revealed that more genes were differentially regulated after 30 min compared to 24 h XB-treated leaves, which correlated with ROS response. Increased xylosidase activity and soluble xylose level after 30 min and 3 h of XB-treatment were observed which might have weakened the DAMP response. However, an increase in total cell wall sugar and a decrease in uronic acid level was observed at both 30 min and 24 h. Additionally, arabinose, rhamnose, and xylose levels were increased in 30 min, and glucose was increased in 24 h compared to mock-treated leaves. The level of jasmonic acid, abscisic acid, auxin, and cytokinin were also affected after XB treatment. Overall, our data revealed that the shortest XOS can act as a DAMP, which triggers the PTI response and alters cell wall composition and hormone level.
Topics: Arabidopsis Proteins; Reactive Oxygen Species; Xylose; Arabidopsis; Cell Wall; Oligosaccharides; Plant Immunity; Gene Expression Regulation, Plant
PubMed: 37991689
DOI: 10.1007/s11103-023-01391-z -
Microbial Cell Factories Aug 2023R. toruloides is an oleaginous yeast, with diverse metabolic capacities and high tolerance for inhibitory compounds abundant in plant biomass hydrolysates. While R....
R. toruloides is an oleaginous yeast, with diverse metabolic capacities and high tolerance for inhibitory compounds abundant in plant biomass hydrolysates. While R. toruloides grows on several pentose sugars and alcohols, further engineering of the native pathway is required for efficient conversion of biomass-derived sugars to higher value bioproducts. A previous high-throughput study inferred that R. toruloides possesses a non-canonical L-arabinose and D-xylose metabolism proceeding through D-arabitol and D-ribulose. In this study, we present a combination of genetic and metabolite data that refine and extend that model. Chiral separations definitively illustrate that D-arabitol is the enantiomer that accumulates under pentose metabolism. Deletion of putative D-arabitol-2-dehydrogenase (RTO4_9990) results in > 75% conversion of D-xylose to D-arabitol, and is growth-complemented on pentoses by heterologous xylulose kinase expression. Deletion of putative D-ribulose kinase (RTO4_14368) arrests all growth on any pentose tested. Analysis of several pentose dehydrogenase mutants elucidates a complex pathway with multiple enzymes mediating multiple different reactions in differing combinations, from which we also inferred a putative L-ribulose utilization pathway. Our results suggest that we have identified enzymes responsible for the majority of pathway flux, with additional unknown enzymes providing accessory activity at multiple steps. Further biochemical characterization of the enzymes described here will enable a more complete and quantitative understanding of R. toruloides pentose metabolism. These findings add to a growing understanding of the diversity and complexity of microbial pentose metabolism.
Topics: Xylose; Arabinose; Pentoses
PubMed: 37537595
DOI: 10.1186/s12934-023-02126-x -
The Separation, Purification, Structure Identification, and Antioxidant Activity of Polysaccharides.Molecules (Basel, Switzerland) Sep 2023In order to investigate the antioxidant activity of polysaccharides, the physicochemical characteristics of purified polysaccharides (EUP, consisting of two fractions,...
In order to investigate the antioxidant activity of polysaccharides, the physicochemical characteristics of purified polysaccharides (EUP, consisting of two fractions, EUP1 and EUP2) were investigated using UV spectrophotometry, high-performance liquid chromatography (HPLC), high-performance gel permeation chromatography (HPGPC), and Fourier transform infrared spectroscopy (FT-IR). This revealed that EUP1 and EUP2 were acidic polysaccharides with an average molecular weight (MW) of 63 and 38 kDa, respectively. EUP1 mainly consisted of L-rhamnose and D-galactose in a molar ratio of 2.05:1, and EUP2 consisted of D-mannose, L-rhamnose, D-galactose, and D-arabinose in a molar ratio of 2.06:1:2.78:1. Furthermore, EUP exhibited considerable antioxidant potential for scavenging hydroxyl, superoxide anion, DPPH, and ABTS radicals. Therefore, EUP can be developed as a potential antioxidant for the functional food or pharmaceutical field.
Topics: Antioxidants; Galactose; Rhamnose; Spectroscopy, Fourier Transform Infrared; Elaeagnaceae; Polysaccharides
PubMed: 37764243
DOI: 10.3390/molecules28186468 -
Microbiology Spectrum Aug 2023Polymyxins are last-line antibiotics employed against multidrug-resistant (MDR) Klebsiella pneumoniae. Worryingly, polymyxin resistance is rapidly on the rise globally....
Polymyxins are last-line antibiotics employed against multidrug-resistant (MDR) Klebsiella pneumoniae. Worryingly, polymyxin resistance is rapidly on the rise globally. Polymyxins initially target lipid A of lipopolysaccharides (LPSs) in the cell outer membrane (OM), causing disorganization and cell lysis. While most studies focus on how genetic variations confer polymyxin resistance, the mechanisms of membrane remodeling and metabolic changes in polymyxin-resistant strains remain unclear, thus hampering the development of effective therapies to treat severe K. pneumoniae infections. In the present study, lipid A profiling, OM lipidomics, genomics, and metabolomics were integrated to elucidate the global mechanisms of polymyxin resistance and metabolic adaptation in a polymyxin-resistant strain (strain S01R; MIC of >128 mg/L) obtained from K. pneumoniae strain S01, a polymyxin-susceptible (MIC of 2 mg/L), New Delhi metallo-β-lactamase (NDM)-producing MDR clinical isolate. Genomic analysis revealed a novel in-frame deletion at position V258 of PhoQ in S01R, potentially leading to lipid A modification with 4-amino-4-deoxy-l-arabinose (L-Ara4N) despite the absence of polymyxin B. Comparative metabolomic analysis revealed slightly elevated levels of energy production and amino acid metabolism in S01R compared to their levels in S01. Exposure to polymyxin B (4 mg/L for S01 and 512 mg/L for S01R) substantially altered energy, nucleotide, and amino acid metabolism and resulted in greater accumulation of lipids in both strains. Furthermore, the change induced by polymyxin B treatment was dramatic at both 1 and 4 h in S01 but only significant at 4 h in S01R. Overall, profound metabolic adaptation was observed in S01R following polymyxin B treatment. These findings contribute to our understanding of polymyxin resistance mechanisms in problematic NDM-producing K. pneumoniae strains and may facilitate the discovery of novel therapeutic targets. Antimicrobial resistance (AMR) is a major threat to global health. The emergence of resistance to the polymyxins that are the last line of defense in so-called Gram-negative "superbugs" has further increased the urgency to develop novel therapies. There are frequent outbreaks of K. pneumoniae infections in hospitals being reported, and polymyxin usage is increasing remarkably. Importantly, the polymyxin-resistant K. pneumoniae strains are imposing more severe consequences to health systems. Using metabolomics, lipid A profiling, and outer membrane lipidomics, our findings reveal (i) changes in the pentose phosphate pathway and amino acid and nucleotide metabolism in a susceptible strain following polymyxin treatment and (ii) how cellular metabolism, lipid A modification, and outer membrane remodeling were altered in K. pneumoniae following the acquisition of polymyxin resistance. Our study provides, for the first time, mechanistic insights into metabolic responses to polymyxin treatment in a multidrug-resistant, NDM-producing K. pneumoniae clinical isolate with acquired polymyxin resistance. Overall, these results will assist in identifying new therapeutic targets to combat and prevent polymyxin resistance.
Topics: Humans; Polymyxins; Polymyxin B; Klebsiella pneumoniae; Lipid A; Anti-Bacterial Agents; Lipid Metabolism; Klebsiella Infections; Microbial Sensitivity Tests
PubMed: 37432123
DOI: 10.1128/spectrum.00852-23 -
International Journal of Molecular... Aug 2023Hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs) are a new class of medications for managing renal anemia in patients with chronic kidney disease (CKD)....
Changes in Metabolomic Profiles Induced by Switching from an Erythropoiesis-Stimulating Agent to a Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibitor in Hemodialysis Patients: A Pilot Study.
Hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs) are a new class of medications for managing renal anemia in patients with chronic kidney disease (CKD). In addition to their erythropoietic activity, HIF-PHIs exhibit multifaceted effects on iron and glucose metabolism, mitochondrial metabolism, and angiogenesis through the regulation of a wide range of HIF-responsive gene expressions. However, the systemic biological effects of HIF-PHIs in CKD patients have not been fully explored. In this prospective, single-center study, we comprehensively investigated changes in plasma metabolomic profiles following the switch from an erythropoiesis-stimulating agent (ESA) to an HIF-PHI, daprodustat, in 10 maintenance hemodialysis patients. Plasma metabolites were measured before and three months after the switch from an ESA to an HIF-PHI. Among 106 individual markers detected in plasma, significant changes were found in four compounds (erythrulose, n-butyrylglycine, threonine, and leucine), and notable but non-significant changes were found in another five compounds (inositol, phosphoric acid, lyxose, arabinose, and hydroxylamine). Pathway analysis indicated decreased levels of plasma metabolites, particularly those involved in phosphatidylinositol signaling, ascorbate and aldarate metabolism, and inositol phosphate metabolism. Our results provide detailed insights into the systemic biological effects of HIF-PHIs in hemodialysis patients and are expected to contribute to an evaluation of the potential side effects that may result from long-term use of this class of drugs.
Topics: Humans; Prolyl Hydroxylases; Pilot Projects; Prolyl-Hydroxylase Inhibitors; Hematinics; Erythropoiesis; Prospective Studies; Procollagen-Proline Dioxygenase; Hypoxia
PubMed: 37628932
DOI: 10.3390/ijms241612752 -
International Journal of Biological... Nov 2023This study investigated the structural and emulsion-stabilizing capacities of alkali-extracted arabinoxylans from corn and wheat bran (CAXs and WAXs). The results...
This study investigated the structural and emulsion-stabilizing capacities of alkali-extracted arabinoxylans from corn and wheat bran (CAXs and WAXs). The results demonstrated that all AXs were mainly composed of arabinose and xylose. WAXs had a higher weight-average molecular weight (Mw, 375-473 KDa) and protein content (3.09-8.68 %) but lower total phenolic acid content (TPC, 1.18-1.91 mg gallic acid equivalents/g) than CAXs; however, CAX stabilized emulsions exhibited smaller and more regular oil droplet size (524-589 nm) and higher absolute value of ζ potential (48-52 mV) compared with WAX stabilized emulsions during storage. Moreover, the increment of NaOH concentration caused a decrease in Mw, protein content, and TPC of CAXs or WAXs and the corresponding CAXs or WAXs emulsions showed bigger and more unstable oil droplets during 14 d storage. The Mw, protein, and TPC were well correlated with their emulsion stability. Furthermore, emulsions stabilized by AXs with low-concentration NaOH could resist better various temperatures, pH, and NaCl. In conclusion, the structural properties of AXs derived from different cereal sources and treated with different concentrations of NaOH varied, leading to differences in their ability to stabilize emulsions. CAXs or WAXs obtained from low-concentration NaOH treatment demonstrated significant potential as highly effective natural emulsifiers.
Topics: Emulsions; Zea mays; Xylans; Dietary Fiber; Alkalies; Molecular Weight; Sodium Hydroxide; Hydrogen-Ion Concentration; Temperature; Triticum
PubMed: 37586632
DOI: 10.1016/j.ijbiomac.2023.126190 -
Vaccine Jul 2023The most common source of foodborne Salmonella infection in humans is poultry eggs and meat, such that prevention of human infection is mostly achieved by vaccination of...
The most common source of foodborne Salmonella infection in humans is poultry eggs and meat, such that prevention of human infection is mostly achieved by vaccination of farm animals. While inactivated and attenuated vaccines are available, both present drawbacks. This study aimed to develop a novel vaccination strategy, which combines the effectiveness of live-attenuated and safety of inactivated vaccines by construction of inducible self-destructing bacteria utilizing toxin-antitoxin (TA) systems. Hok-Sok and CeaB-CeiB toxin-antitoxin systems were coupled with three induction systems aimed for activating cell killing upon lack of arabinose, anaerobic conditions or low concentration of metallic di-cations. The constructs were transformed into a pathogenic Salmonella enterica serovar Enteritidis strain and bacteria elimination was evaluated in vitro under specific activating conditions and in vivo following administration to chickens. Four constructs induced bacterial killing under the specified conditions, both in growth media and within macrophages. Cloacal swabs of all chicks orally administered transformed bacteria had no detectable levels of bacteria within 9 days of inoculation. By day ten, no bacteria were identified in the spleen and liver of most birds. Antibody immune response was raised toward TA carrying Salmonella which resembled response toward the wildtype bacteria. The constructs described in this study led to self-destruction of virulent Salmonella enteritidis both in vitro and in inoculated animals within a period which is sufficient for the induction of a protective immune response. This system may serve as a safe and effective live vaccine platform against Salmonella as well as other pathogenic bacteria.
Topics: Animals; Humans; Antitoxins; Chickens; Salmonella Infections, Animal; Salmonella enteritidis; Toxins, Biological; Vaccination; Salmonella Vaccines; Poultry Diseases; Vaccines, Attenuated
PubMed: 37400285
DOI: 10.1016/j.vaccine.2023.06.074