Alginate lyases have countless potential for application in industries and medicine particularly as an appealing biocatalyst for the production of biofuels and bioactive oligosaccharides. Solid-state fermentation (SSF) allows improved production of enzymes and consumes less energy compared to submerged fermentation. Seaweeds can serve as the most promising biomass for the production of biochemicals. Alginate present in the seaweed can be used by alginate lyase-producing bacteria to support growth and can secrete alginate lyase. In this perspective, the current study was directed on the bioprocessing of brown seaweeds for the production of alginate lyase using marine bacterial isolate. A novel alginate-degrading marine bacterium Enterobacter tabaci RAU2C which was previously isolated in the laboratory was used for the production of alginate lyase using Sargassum swartzii as a low-cost solid substrate. Process parameters such as inoculum incubation period and moisture content were optimized for alginate lyase production. SSF resulted in 33.56 U/mL of alginate lyase under the static condition maintained with 75% moisture after 4 days. Further, the effect of different buffers, pH, and temperature on alginate lyase activity was also analyzed. An increase in alginate lyase activity was observed with an increase in moisture content from 60 to 75%. Maximum enzyme activity was perceived with phosphate buffer at pH 7 and 37 °C. Further, the residual biomass after SSF could be employed as biofertilizer for plant growth promotion based on the preliminary analysis. To our knowledge, this is the first report stating the usage of seaweed biomass as a substrate for the production of alginate lyase using solid-state fermentation.
- MeSH
- Alginates * metabolism MeSH
- Biomass MeSH
- Enterobacter * metabolism enzymology isolation & purification growth & development MeSH
- Fermentation * MeSH
- Hydrogen-Ion Concentration MeSH
- Glucuronic Acid metabolism MeSH
- Seaweed * microbiology MeSH
- Phaeophyceae microbiology MeSH
- Polysaccharide-Lyases * metabolism MeSH
- Sargassum * microbiology metabolism MeSH
- Temperature MeSH
- Publication type
- Journal Article MeSH
Luteolin and naringenin are flavonoids found in various foods/beverages and present in certain dietary supplements. After a high intake of these flavonoids, their sulfate and glucuronide conjugates reach micromolar concentrations in the bloodstream. Some pharmacokinetic interactions of luteolin and naringenin have been investigated in previous studies; however, only limited data are available in regard to their metabolites. In this study, we aimed to investigate the interactions of the sulfate and glucuronic acid conjugates of luteolin and naringenin with human serum albumin, cytochrome P450 (CYP2C9, 2C19, and 3A4) enzymes, and organic anion transporting polypeptide (OATP1B1 and OATP2B1) transporters. Our main findings are as follows: (1) Sulfate conjugates formed more stable complexes with albumin than the parent flavonoids. (2) Luteolin and naringenin conjugates showed no or only weak inhibitory action on the CYP enzymes examined. (3) Certain conjugates of luteolin and naringenin are potent inhibitors of OATP1B1 and/or OATP2B1 enzymes. (4) Conjugated metabolites of luteolin and naringenin may play an important role in the pharmacokinetic interactions of these flavonoids.
- MeSH
- Cytochrome P-450 CYP3A * metabolism MeSH
- Cytochrome P-450 CYP2C19 metabolism MeSH
- Cytochrome P-450 CYP2C9 metabolism MeSH
- Flavonoids pharmacology MeSH
- Glucuronides MeSH
- Humans MeSH
- Serum Albumin, Human metabolism MeSH
- Luteolin pharmacology MeSH
- Organic Anion Transporters * metabolism MeSH
- Sulfates metabolism MeSH
- Cytochrome P-450 Enzyme System metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Cyanobacteria produce a wide range of metabolites of interest for industrial or medical use. The cultivation of freshwater Nostoc cf. linckia yielded 5.4 g/L of a crude exopolysaccharide (cEPS) with a molecular weight of 1.31 × 105 g/mol. Ion-exchange chromatography of cEPS yielded two dominant fractions, EPS-1 and EPS-2, differing in molecular weight. The lower molecular weight fraction (EPS-1) was subjected to structural studies. Results of chemical and spectroscopic analyses showed that three of the four dominant sugars, glucose, galactose and xylose are 1,4-linked in the backbone in the following order: [→4)-β-D-Xylp-(1 → 4)-β-D-Glcp-(1 → 4)-α-D-Galp-(1 → 4)-β-D-Glcp-(1→]n. Terminal mannose residues were identified as side chains linked at C3 of every third backbone xylose and every second glucose is branched at C6 by 3-O-lactyl-β-D-glucuronic acid (nosturonic acid). Antioxidant properties of EPS were tested using two in vitro methods. Both assays showed that the cEPS was more active than purified EPS-1 and EPS-2 fractions and deproteinized EPS.
- MeSH
- Antioxidants chemistry MeSH
- Polysaccharides, Bacterial analysis chemistry MeSH
- Galactose chemistry MeSH
- Glucose chemistry MeSH
- Glucuronic Acid chemistry MeSH
- Magnetic Resonance Spectroscopy methods MeSH
- Molecular Structure MeSH
- Molecular Weight MeSH
- Nostoc chemistry MeSH
- Xylose chemistry MeSH
- Publication type
- Journal Article MeSH
Pochopení cest metabolizace inhibitorů SGLT2 (gliflozinů) je jednou z podmínek bezpečné terapie těmito léčivy. Glifloziny se v organismu metabolizují prakticky jen glukuronidací, což je velmi neobvyklé a lékaři i farmaceuti s tím musejí počítat. Největší část všech gliflozinů se metabolizuje cestou UGT1A9, v případě dapagliflozinu je tato cesta rozhodující, naopak empagliflozin je metabolizován hned čtyřmi izoenzymy UGT, což se projevuje zanedbatelným vlivem jejich polymorfismů a lékových interakcí. Glukuronidy vzniklé metabolizací gliflozinů jsou dobře rozpustné ve vodě, a jsou proto eliminovány ledvinami aktivním transportem. Míra glukuronidace jednotlivých gliflozinů je různá, nejnižší je v případě empagliflozinu a nejvyšší u dapagliflozinu.
Understanding the metabolism pathways of SGLT2 inhibitors (gliflozins) is one of the conditions for safe therapy with these drugs. Gliflozins are metabolized in the body practically only by glucuronidation, which is very unusual. Both doctors and pharmacists must count it in. Most gliflozins are metabolized by UGT1A9. In the case of dapagliflozin, this is the decisive pathway, whereas empagliflozin is metabolized by four UGT isoenzymes, which manifests as a negligible effect of their polymorphisms and drug interactions. Glucuronides formed by the metabolism of gliflozins are readily soluble in water and are therefore eliminated by active renal transport. The degree of glucuronidation of individual gliflozins is different; the lowest is in the case of empagliflozin and the highest in the case of dapagliflozin.
Deoxynivalenol (DON) and its modified forms, including DON-3-glucoside (DON-3G), pose a major agricultural and food safety issue in the world. Their metabolites are relatively well-characterized; however, their metabolizing enzymes have not been fully explored. UDP-glucuronosyltransferases, 3-O-acetyltransferase, and glutathione S-transferase are involved in the formation of DON-glucuronides, 3-acetyl-DON, and DON-glutathione, respectively. There are interindividual differences in the metabolism of these toxins, including variation with respect to sex. Furthermore, interspecies differences in DON metabolism have been revealed, including differences in the major metabolites of DON, the role of de-acetylation, and the hydrolysis of DON-3G. In this review, we summarized the major enzymes involved in metabolizing DON to its modified forms, focusing on the differences in metabolism of DON and its modified forms between individuals and species. This work provides important insight into the toxicity of DON and its derivatives in humans and animals, and provides scientific basis for the development of safer and more efficient biological detoxification methods.
- MeSH
- Glucuronides metabolism MeSH
- Glucuronosyltransferase metabolism MeSH
- Hydrolysis MeSH
- Humans MeSH
- Inactivation, Metabolic * MeSH
- Trichothecenes * chemistry metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
Mycophenolic acid (MPA) has become a cornerstone of immunosuppressive therapy, in particular for transplant patients. In the gastrointestinal tract, the liver and the kidney, MPA is mainly metabolized into phenyl-β-d glucuronide (MPAG). Knowledge about the interactions between MPA/MPAG and membrane transporters is still fragmented. The aim of the present study was to explore these interactions with the basolateral hepatic MRP4 transporter. The inhibition of the MRP4-driven transport by various drugs which can be concomitantly prescribed was also evaluated. In vitro experiments using vesicles overexpressing MRP4 showed an ATP-dependent transport of MPAG driven by MRP4 (Michaelis-Menten constant of 233.9 ± 32.8 µM). MPA was not effluxed by MRP4. MRP4-mediated transport of MPAG was inhibited (from -43% to -84%) by ibuprofen, cefazolin, cefotaxime and micafungin. An in silico approach based on molecular docking and molecular dynamics simulations rationalized the mode of binding of MPAG to MRP4. The presence of the glucuronide moiety in MPAG was highlighted as key, being prone to make electrostatic and H-bond interactions with specific residues of the MRP4 protein chamber. This explains why MPAG is a substrate of MRP4 whereas MPA is not.
- MeSH
- Biological Transport MeSH
- Glucuronides metabolism MeSH
- Hepatocytes metabolism MeSH
- Liver metabolism MeSH
- Mycophenolic Acid analogs & derivatives metabolism MeSH
- Kidney metabolism MeSH
- Humans MeSH
- Membrane Transport Proteins metabolism MeSH
- Multidrug Resistance-Associated Proteins metabolism MeSH
- Molecular Docking Simulation MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Silybum marianum (milk thistle) is a medicinal plant used for producing the hepatoprotective remedy silymarin. Its main bioactive constituents, including silybin and related flavonolignans, can be metabolized directly by phase II conjugation reactions. This study was designed to identify UDP-glucuronosyltransferases (UGTs) involved in the glucuronidation of six silymarin flavonolignans, namely silybin A, silybin B, isosilybin A, isosilybin B, silychristin, and silydianin. UHPLC-MS analyses showed that all of the tested compounds, both individually and in silymarin, were glucuronidated by human liver microsomes, and that glucuronidation was the main metabolic transformation in human hepatocytes. Further, each compound was glucuronidated by multiple recombinant human UGT enzymes. UGTs 1A1, 1A3, 1A8 and 1A9 were able to conjugate all of the tested flavonolignans, and some of them were also metabolized by UGTs 1A6, 1A7, 1A10, 2B7 and 2B15. In contrast, no glucuronides were produced by UGTs 1A4, 2B4, 2B10 and 2B17. With silymarin, we found that UGT1A1 and, to a lesser extent UGT1A9, were primarily responsible for the glucuronidation of the flavonolignan constituents. It is concluded that the metabolism of silymarin flavonolignans may involve multiple UGT enzymes, of which UGT1A1 appears to play the major role in the glucuronidation. These results may be relevant for future research on the metabolism of flavonolignans in humans.
- MeSH
- Adult MeSH
- Flavonolignans metabolism MeSH
- Glucuronides metabolism MeSH
- Glucuronosyltransferase metabolism MeSH
- Hepatocytes metabolism MeSH
- Microsomes, Liver metabolism MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Silybum marianum metabolism MeSH
- Silybin metabolism MeSH
- Silymarin analogs & derivatives metabolism MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
Complex structure of cyanobacterium Nostoc sp. exopolysaccharide (EPS), with apparent molecular weight 214 × 103 g/mol, can be deduced from its composition. Chemical and NMR analyses found four dominant sugar monomers, namely (1 → 4)-linked α-l-arabinopyranose, β-d-glucopyranose, β-d-xylopyranose and (1 → 3)-linked β-d-mannopyranose, two different uronic acids and a lactyl group, with (1 → 4,6)-linked β-d-glucopyranose as the only branch point suggest a complex structure of this polymer. The dominant uronic acid is α-linked, but it remained unidentified. β-d-Glucuronic acid was present in lower amount. Their position as well as that of lactyl remained undetermined too. Different doses of orally administered EPS in guinea pigs evoked a significant decrease in cough effort and a decrease in airway reactivity. The antitussive efficacy and bronchodilator effect of higher doses of EPS were found to be similar to that of the antitussive drug codeine and the antiasthmatic salbutamol. Without significant cytotoxicity on the RAW 264.7 cells, EPS stimulated the macrophage cells to produce pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and prostaglandins (PGs) and nitric oxide (NO) via induction of COX-2 and iNOS expression, respectively, suggesting that this biopolymer potentiates an early innate immune response and can therefore be used as a new immune modulator.
- MeSH
- Albuterol pharmacology MeSH
- Polysaccharides, Bacterial chemistry pharmacology MeSH
- Biopolymers chemistry MeSH
- Bronchodilator Agents pharmacology MeSH
- Cell Line MeSH
- Cytokines metabolism MeSH
- Interleukin-6 metabolism MeSH
- Cough drug therapy MeSH
- Codeine pharmacology MeSH
- Glucuronic Acid chemistry MeSH
- Uronic Acids chemistry MeSH
- Macrophages drug effects metabolism MeSH
- Guinea Pigs MeSH
- Mice MeSH
- Nostoc metabolism MeSH
- Nitric Oxide metabolism MeSH
- RAW 264.7 Cells MeSH
- Cyanobacteria metabolism MeSH
- Tumor Necrosis Factor-alpha metabolism MeSH
- Animals MeSH
- Check Tag
- Guinea Pigs MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
The work is focused on the development of microspheres based on the combination of two polysaccharides; chitosan and alginic acid with the aim to allocate, hold, release and protect environmentally sensible molecules. The microspheres were prepared using a solvent-free, low cost and scalable approach and two enzymes; trypsin and protease from Aspergillus Oryzae have been used as a model to evaluate the microspheres peculiarities. The proteins were encapsulated during the microspheres preparation. The relationship between the polysaccharides weight ratio and the morphology, stability and ability of the carrier to allocate the enzymes has been evaluated. The enzymatic activity and the release kinetics were assessed in different conditions to assess the impact of the external environment. Obtained results demonstrate the efficacy of the prepared microspheres to preserve the activity of relevant bioactive compounds which are highly relevant in food, cosmetic and pharmaceutic, but the application is limited due to their high sensibility.
- MeSH
- Aspergillus oryzae enzymology MeSH
- NIH 3T3 Cells MeSH
- Chitosan chemistry toxicity MeSH
- Enzymes, Immobilized chemistry metabolism MeSH
- Hydrogen-Ion Concentration MeSH
- Alginic Acid chemistry toxicity MeSH
- Humans MeSH
- Microspheres * MeSH
- Mice MeSH
- Materials Testing MeSH
- Capsules MeSH
- Trypsin chemistry metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
