26 results on '"Bridiau N"'
Search Results
2. Heparanase targeted for regulation of breast tumoral angiogenesis using structural analogues of heparan sulfate
- Author
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Poupard, N., Badarou, P., Bridiau, N., Kieda, Claudine, Piot, J-M., Maugard, T., Fruitier-Arnaudin, I., Frapart, Isabelle, Institut de Chimie de Clermont-Ferrand (ICCF), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-SIGMA Clermont (SIGMA Clermont)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), LIttoral ENvironnement et Sociétés (LIENSs), and La Rochelle Université (ULR)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
[SDV] Life Sciences [q-bio] ,[SDV]Life Sciences [q-bio] ,ComputingMilieux_MISCELLANEOUS - Abstract
International audience
- Published
- 2016
3. β-Galactosidase Catalyzed Selective Galactosylation of Aromatic Compounds
- Author
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Bridiau, N., primary, Taboubi, S., additional, Marzouki, N., additional, Legoy, M.D., additional, and Maugard, T., additional
- Published
- 2006
- Full Text
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4. Semi-pilot scale-up of a continuous packed-bed bioreactor system developed for the lipase-catalyzed production of pseudo-ceramides
- Author
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Le Joubioux Florian, Bridiau Nicolas, Graber Marianne, and Maugard Thierry
- Subjects
pseudo-ceramide ,continuous bioprocess ,packed-bed bioreactor ,lipase ,semi-pilot scale ,Oils, fats, and waxes ,TP670-699 - Abstract
Ceramides are sphingolipid compounds that are very attractive as active components in both the pharmaceutical and the cosmetic industries. In this study, the synthesis of 1-O,3-N-diacyl 3-amino-1,2-propanediol-type pseudo-ceramides was developed at the semi-pilot scale, starting from a two-step continuous enzymatic process with immobilized Candida antarctica lipase B (Novozym® 435) in a packed-bed bioreactor, previously optimized at the laboratory scale. This process involved the selective N-acylation of 3-amino-1,2-propanediol (step 1), followed by the selective O-acylation of the N-acyl 3-amino-1,2-propanediol synthesized in the first step, with various fatty acids as acyl donors, to produce N,O-diacyl 3-amino-1,2-propanediol-type pseudo-ceramides (step 2). Under partially optimized operating conditions, high synthesis yields and production rates were obtained, within the ranges 76–92% and 3.7–4.6 g h−1 (step 1), or 23–36% and 1–1.4 g h−1 (step 2), respectively, depending on the fatty acids used as acyl donors. The overall synthesis yields varied from 20 to 33%: the best yield was obtained using palmitic acid and lauric acid as first and second acyl donors, respectively. Together with the high production rates also obtained with these acyl donors, this confirms that this two-step process has great potential for the production of differently functionalized 1-O,3-N-diacyl 3-amino-1,2-propanediol-type pseudo-ceramides on an industrial scale.
- Published
- 2017
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5. Exopolysaccharide from marine microalgae belonging to the Glossomastix genus: fragile gel behavior and suspension stability.
- Author
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Dulong V, Rihouey C, Gaignard C, Bridiau N, Gourvil P, Laroche C, Pierre G, Varacavoudin T, Probert I, Maugard T, Michaud P, Picton L, and Le Cerf D
- Subjects
- Polysaccharides chemistry, Alginates, Photobioreactors, Polysaccharides, Bacterial, Microalgae
- Abstract
With the aim to find new polysaccharides of rheological interest with innovated properties, rhamnofucans produced as exopolysaccharides (EPS) in a photobioreactor (PBR) and an airlift bioreactor (ABR) by the marine microalgae Glossomastix sp. RCC3707 and RCC3688 were fully studied. Chemical characterizations have been conducted (UHPLC - MS HR). Analyses by size-exclusion chromatography (SEC) coupled online with a multiangle light scattering detector (MALS) and a differential refractive index detector showed the presence of large structures with molar masses higher than 10
6 g.mol-1 . The rheological studies of these EPS solutions, conducted at different concentrations and salinities, have evidenced interesting and rare behavior characteristic of weak and fragile hydrogels i.e. gel behavior with very low elastic moduli (between 10-2 and 10 Pa) and yield stresses (between 10-2 and 2 Pa) according to the EPS source, concentration, and salinity. These results were confirmed by diffusing wave spectroscopy. Finally, as one of potential application, solutions of EPS from Glossomastix sp. have evidenced very good properties as anti-settling stabilizers, using microcrystalline cellulose particles as model, studied by multiple light scattering (MLS) with utilization in cosmetic or food industry. Compared to alginate solution with same viscosity for which sedimentation is observed over few hours, microalgae EPS leads to a stable suspension over few days.- Published
- 2024
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6. Role of some structural features in EPS from microalgae stimulating collagen production by human dermal fibroblasts.
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Toucheteau C, Deffains V, Gaignard C, Rihouey C, Laroche C, Pierre G, Lépine O, Probert I, Le Cerf D, Michaud P, Arnaudin-Fruitier I, Bridiau N, and Maugard T
- Subjects
- Humans, Collagen, Fibroblasts, Sulfates, Matrix Metalloproteinase 1, Microalgae
- Abstract
Exopolysaccharides (EPS) from the microalgae Porphyridium cruentum , Chrysotila dentata , Pavlova sp., Diacronema sp., Glossomastix sp., Phaeodactylum tricornutum, and Synechococcus sp. were isolated and depolymerized. First, EPS were submitted to a high pressure pre-treatment step, followed by a solid acid-catalyzed hydrolysis step carried out in a batch or recycle fixed-bed reactor, using a strong acidic cation-exchange resin. Twenty-eight different EPS forms were thus obtained. After characterization of their main structural features (weight- and number-averaged molecular weight, polydispersity index, sulfate and uronic acid contents), we investigated the structure-function relationship of their pro-collagen activity. We found that native microalgae EPS were able to inhibit until 27% of human matrix metalloproteinase-1 (MMP-1) activity while the depolymerized forms were able to enhance collagen production by two different human fibroblast lines, used as cell models due to their major role in dermal collagen biosynthesis. The most active EPS forms, obtained by depolymerization in the recycle fixed-bed reactor of D. ennorea and Glossomastix sp. EPS, led to 390% increase in collagen production. Finally, principal component (PCA) and Pearson analyses indicated that MMP-1 inhibition was strongly correlated to the sulfate group content of EPS whereas collagen production by fibroblasts was mostly related to their proportion of low molecular weight polysaccharides (<10 kDa). Uronic acid content of EPS was also shown essential but only if the size of EPS was reduced in the first place. Altogether, these results gave new insights of the dermo-cosmetic potential of microalgae EPS as well as the key parameters of their activity.
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- 2023
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7. A Screening Approach to Assess the Impact of Various Commercial Sources of Crude Marine λ-Carrageenan on the Production of Oligosaccharides with Anti-heparanase and Anti-migratory Activities.
- Author
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Manseur C, Groult H, Porta M, Bodet PE, Mersni-Achour R, Petit R, Ali-Moussa S, Musnier B, Le Cerf D, Varacavoudin T, Haddad O, Sutton A, Leal CEY, Alencar-Filho EB, Piot JM, Bridiau N, Maugard T, and Fruitier-Arnaudin I
- Subjects
- Humans, Carrageenan pharmacology, Carrageenan chemistry, Oligosaccharides pharmacology, Oligosaccharides chemistry, Hydrogen Peroxide pharmacology, Neoplasms
- Abstract
Oligosaccharides derived from λ-carrageenan (λ-COs) are gaining interest in the cancer field. They have been recently reported to regulate heparanase (HPSE) activity, a protumor enzyme involved in cancer cell migration and invasion, making them very promising molecules for new therapeutic applications. However, one of the specific features of commercial λ-carrageenan (λ-CAR) is that they are heterogeneous mixtures of different CAR families, and are named according to the thickening-purpose final-product viscosity which does not reflect the real composition. Consequently, this can limit their use in a clinical applications. To address this issue, six commercial λ-CARs were compared and differences in their physiochemical properties were analyzed and shown. Then, a H
2 O2 -assisted depolymerization was applied to each commercial source, and number- and weight-averaged molar masses (Mn and Mw ) and sulfation degree (DS) of the λ-COs produced over time were determined. By adjusting the depolymerization time for each product, almost comparable λ-CO formulations could be obtained in terms of molar masses and DS, which ranged within previously reported values suitable for antitumor properties. However, when the anti-HPSE activity of these new λ-COs was screened, small changes that could not be attributed only to their small length or DS changes between them were found, suggesting a role of other features, such as differences in the initial mixture composition. Further structural MS and NMR analysis revealed qualitative and semi-quantitative differences between the molecular species, especially in the proportion of the anti-HPSE λ-type, other CARs types and adjuvants, and it also showed that H2 O2 -based hydrolysis induced sugar degradation. Finally, when the effects of λ-COs were assessed in an in vitro migration cell-based model, they seemed more related to the proportion of other CAR types in the formulation than to their λ-type-dependent anti-HPSE activity.- Published
- 2023
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8. Ulva intestinalis Protein Extracts Promote In Vitro Collagen and Hyaluronic Acid Production by Human Dermal Fibroblasts.
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Bodin J, Adrien A, Bodet PE, Dufour D, Baudouin S, Maugard T, and Bridiau N
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- Amino Acids chemistry, Cell Survival drug effects, Cells, Cultured, Chromatography, High Pressure Liquid, Humans, Hydrolysis, Peptides chemistry, Peptides pharmacology, Plant Extracts chemistry, Plant Proteins chemistry, Protein Biosynthesis drug effects, Collagen biosynthesis, Fibroblasts drug effects, Fibroblasts metabolism, Hyaluronic Acid biosynthesis, Plant Extracts pharmacology, Plant Proteins pharmacology, Skin cytology, Ulva chemistry
- Abstract
With the increase in life expectancy, reducing the visible signs of skin aging has become a major issue. A reduction in collagen and hyaluronic acid synthesis by fibroblasts is a feature of skin aging. The green seaweed, Ulva intestinalis , is an abundant and rich source of nutrients, especially proteins and peptides. The aim of this study was to assess the potential cosmetic properties of a protein fraction from Ulva intestinalis (PROT-1) containing 51% of proteins and 22% of polysaccharides, and its enzymatic peptide hydrolysates on human dermal fibroblasts. PROT-1 was extracted using a patented acid- and solvent-free process (FR2998894 (B1)). The biochemical characterization and chromatographic analysis showed a main set of proteins (25 kDa). To demonstrate the anti-aging potential of PROT-1, fibroblast proliferation and collagen and hyaluronic acid production were assessed on fibroblast cell lines from donors aged 20 years (CCD-1059Sk) and 46 years (CCD-1090Sk). PROT-1 induced a significant increase in collagen and hyaluronic acid production per cell, and a reduction in cell proliferation without increasing cell mortality. These effects were reversed after protein hydrolysis of PROT-1, showing the central role of proteins in this promising anti-aging property.
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- 2020
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9. Anticoagulant Activity of Sulfated Ulvan Isolated from the Green Macroalga Ulva rigida .
- Author
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Adrien A, Bonnet A, Dufour D, Baudouin S, Maugard T, and Bridiau N
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- Anticoagulants chemistry, Anticoagulants isolation & purification, Blood Coagulation Tests, Enoxaparin pharmacology, Heparin pharmacology, Humans, Plasma drug effects, Anticoagulants pharmacology, Fibroblasts drug effects, Ulva chemistry
- Abstract
(1) Background: Brown and red algal sulfated polysaccharides have been widely described as anticoagulant agents. However, data on green algae, especially on the Ulva genus, are limited. This study aimed at isolating ulvan from the green macroalga Ulva rigida using an acid- and solvent-free procedure, and investigating the effect of sulfate content on the anticoagulant activity of this polysaccharide. (2) Methods: The obtained ulvan fraction was chemically sulfated, leading to a doubling of the polysaccharide sulfate content in a second ulvan fraction. The potential anticoagulant activity of both ulvan fractions was then assessed using different assays, targeting the intrinsic and/or common (activated partial thromboplastin time), extrinsic (prothrombin time), and common (thrombin time) pathways, and the specific antithrombin-dependent pathway (anti-Xa and anti-IIa), of the coagulation cascade. Furthermore, their anticoagulant properties were compared to those of commercial anticoagulants: heparin and Lovenox
® . (3) Results: The anticoagulant activity of the chemically-sulfated ulvan fraction was stronger than that of Lovenox® against both the intrinsic and extrinsic coagulation pathways. (4) Conclusion: The chemically-sulfated ulvan fraction could be a very interesting alternative to heparins, with different targets and a high anticoagulant activity.- Published
- 2019
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10. λ-Carrageenan Oligosaccharides of Distinct Anti-Heparanase and Anticoagulant Activities Inhibit MDA-MB-231 Breast Cancer Cell Migration.
- Author
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Groult H, Cousin R, Chot-Plassot C, Maura M, Bridiau N, Piot JM, Maugard T, and Fruitier-Arnaudin I
- Subjects
- Anticoagulants chemistry, Antineoplastic Agents chemistry, Breast Neoplasms drug therapy, Breast Neoplasms pathology, Carrageenan chemistry, Cell Line, Tumor, Cell Movement drug effects, Drug Screening Assays, Antitumor, Enzyme Assays, Female, Glucuronidase metabolism, Heparitin Sulfate metabolism, Humans, Oligosaccharides chemistry, Anticoagulants pharmacology, Antineoplastic Agents pharmacology, Carrageenan pharmacology, Glucuronidase antagonists & inhibitors, Oligosaccharides pharmacology
- Abstract
In tumor development, the degradation of heparan sulfate (HS) by heparanase (HPSE) is associated with cell-surface and extracellular matrix remodeling as well as the release of HS-bound signaling molecules, allowing cancer cell migration, invasion and angiogenesis. Because of their structural similarity with HS, sulfated polysaccharides are considered a promising source of molecules to control these activities. In this study, we used a depolymerisation method for producing λ-carrageenan oligosaccharides (λ-CO), with progressive desulfation over time. These were then used to investigate the influence of polymeric chain length and degree of sulfation (DS) on their anti-HPSE activity. The effects of these two features on λ-CO anticoagulant properties were also investigated to eliminate a potential limitation on the use of a candidate λ-CO as a chemotherapeutic agent. HPSE inhibition was mainly related to the DS of λ-CO, however this correlation was not complete. On the other hand, both chain length and DS modulated λ-CO activity for factor Xa and thrombin IIa inhibition, two enzymes that are involved in the coagulation cascade, and different mechanisms of inhibition were observed. A λ-carrageenan oligosaccharide of 5.9 KDa was identified as a suitable anticancer candidate because it displayed one of the lowest anticoagulant properties among the λ-CO produced, while showing a remarkable inhibitory effect on MDA-MB-231 breast cancer cell migration.
- Published
- 2019
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11. Family of Bioactive Heparin-Coated Iron Oxide Nanoparticles with Positive Contrast in Magnetic Resonance Imaging for Specific Biomedical Applications.
- Author
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Groult H, Poupard N, Herranz F, Conforto E, Bridiau N, Sannier F, Bordenave S, Piot JM, Ruiz-Cabello J, Fruitier-Arnaudin I, and Maugard T
- Subjects
- Animals, Contrast Media chemistry, Female, HEK293 Cells, Humans, Metal Nanoparticles adverse effects, Mice, Mice, Inbred C57BL, Ferric Compounds chemistry, Heparin chemistry, Magnetic Resonance Angiography methods, Metal Nanoparticles chemistry
- Abstract
Unfractionated heparin (UFH) and low-molecular-weight heparins (LMWH) are well-known for their anticoagulant properties. There is also currently a growing interest in using LMWH in targeted cancer therapy. In particular, several types inhibit heparanase, a key enzyme overexpressed in the tumor microenvironment that promotes angiogenesis progression and metastasis spreading. Here, we propose iron oxide nanoparticles (HEP-IONP) coated with different heparins of distinct anticoagulant/anti-heparanase activity ratios and suitable for positive contrast in magnetic resonance imaging. As a proof of concept, magnetic resonance angiography (MRA) was conducted in mice up to 3 h after intravenous administration. This new IONP-based positive contrast appropriate for clinic together with the long vascular circulating times can enable innovative theranostic applications if combined with the various bioactivities of the heparins. Indeed, we showed, using advanced in vitro tests, how HEP-IONP anticoagulant or anti-heparanase activities were maintained depending on the heparin species used for the coating. Overall, the study allowed presenting an IONP coated with a commercial LMWH (Lovenox) suggested as a theranostic translational probe for MRA diagnostic and treatment of thrombosis, and an antitumor IONP coated with a specific depolymerized heparin to be used in targeted therapy and diagnostic modalities.
- Published
- 2017
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12. Evaluation of the anticoagulant potential of polysaccharide-rich fractions extracted from macroalgae.
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Adrien A, Dufour D, Baudouin S, Maugard T, and Bridiau N
- Subjects
- Anticoagulants chemistry, Blood Coagulation drug effects, Drug Evaluation, Preclinical methods, Heparin pharmacology, Humans, Partial Thromboplastin Time, Anticoagulants pharmacology, Phaeophyceae chemistry, Polysaccharides chemistry, Polysaccharides pharmacology, Seaweed chemistry
- Abstract
The aim of this study was to evaluate the potential anticoagulant activity of sulphated polysaccharide-containing extracts of six french edible marine macroalgae. Aqueous extracts of brown (Himanthalia elongata, Laminaria digitata, Ascophyllum nodosum, Fucus vesiculosus), green (Ulva lactuca) and red (Chondrus crispus) macroalgae were prepared and their biochemical properties were determined, including major biomolecules, sulphate and ash contents. The anticoagulant activity of each extract was investigated using different scales from the specific antithrombin-dependent pathway (anti-Xa and anti-IIa) to the intrinsic and/or common (Activated Partial Thromboplastin Time, APTT), extrinsic (Prothrombin Time, PT) or common (Thrombin Time, TT) anticoagulant pathways, and compared with those of commercial anticoagulants, heparin and Lovenox®. Laminaria digitata, Fucus vesiculosus and Chondrus crispus extracts showed a significant APTT anticoagulant capacity, only 5-fold lower than that of Lovenox®, which is a pure low molecular weight heparin used as an anticoagulant agent to prevent pulmonary embolism in patients undergoing surgery.
- Published
- 2017
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13. Production of heparin and λ-carrageenan anti-heparanase derivatives using a combination of physicochemical depolymerization and glycol splitting.
- Author
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Poupard N, Groult H, Bodin J, Bridiau N, Bordenave-Juchereau S, Sannier F, Piot JM, Fruitier-Arnaudin I, and Maugard T
- Subjects
- Glycols, Carrageenan chemistry, Glucuronidase antagonists & inhibitors, Heparin chemistry
- Abstract
Strongly associated with tumor angiogenesis and metastasis, the enzyme heparanase is an endo-β-d-glucuronidase which is overexpressed in the tumor microenvironment. Its inhibition could be one of the most promising anti-angiogenic approaches to date. Although heparin is known as a good heparanase inhibitor, it also possesses major anticoagulant properties that may be incompatible with its use as an anti-angiogenic agent, hence the considerable interest for other sources of sulfated polysaccharides. Recent investigations point to λ-carrageenans, highly sulfated galactans with a tremendous potential that are found in red algae. This study describes the production of low-molecular-weight (LMW) heparins and λ-carrageenans, using a combination of glycol splitting and ultrasonically-assisted radical hydrolysis using hydrogen-peroxide. The structural characteristics, as well as the anticoagulant and antiheparanase activities of the resulting products were assessed. The best candidate was a LMW glycol-split λ-carrageenan that displayed major anti-heparanase properties, with an IC
50 of 7.32ng/mL and a close-to-zero anticoagulant activity., (Copyright © 2017 Elsevier Ltd. All rights reserved.)- Published
- 2017
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14. Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent.
- Author
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Poupard N, Badarou P, Fasani F, Groult H, Bridiau N, Sannier F, Bordenave-Juchereau S, Kieda C, Piot JM, Grillon C, Fruitier-Arnaudin I, and Maugard T
- Subjects
- Cell Line, Cell Line, Tumor, Extracellular Matrix drug effects, Heparin pharmacology, Humans, MCF-7 Cells, Neoplasms drug therapy, Tumor Microenvironment drug effects, Angiogenesis Inhibitors pharmacology, Carrageenan pharmacology, Endothelial Cells drug effects, Glucuronidase antagonists & inhibitors, Neovascularization, Pathologic drug therapy
- Abstract
Heparanase is overexpressed by tumor cells and degrades the extracellular matrix proteoglycans through cleavage of heparan sulfates (HS), allowing pro-angiogenic factor release and thus playing a key role in tumor angiogenesis and metastasis. Here we propose new HS analogs as potent heparanase inhibitors: Heparin as a positive control, Dextran Sulfate, λ-Carrageenan, and modified forms of them obtained by depolymerization associated to glycol splitting (RD-GS). After heparanase activity assessment, 11 kDa RD-GS-λ-Carrageenan emerged as the most effective heparanase inhibitor with an IC
50 of 7.32 ng/mL compared to 10.7 ng/mL for the 16 kDa unfractionated heparin. The fractionated polysaccharides were then tested in a heparanase-rich medium-based in vitro model, mimicking tumor microenvironment, to determine their effect on microvascular endothelial cells (HSkMEC) angiogenesis. As a preliminary study, we identified that under hypoxic and nutrient poor conditions, MCF-7 cancer cells released much more mature heparanase in their supernatant than in normal conditions. Then a MatrigelTM assay using HSkMEC cultured under hypoxic conditions in the presence (or not) of this heparanase-rich supernatant was realized. Adding heparanase-rich media strongly enhanced angiogenic network formation with a production of twice more pseudo-vessels than with the control. When sulfated polysaccharides were tested in this angiogenesis assay, RD-GS-λ-Carrageenan was identified as a promising anti-angiogenic agent.- Published
- 2017
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15. Pilot production of ulvans from Ulva sp. and their effects on hyaluronan and collagen production in cultured dermal fibroblasts.
- Author
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Adrien A, Bonnet A, Dufour D, Baudouin S, Maugard T, and Bridiau N
- Subjects
- Cells, Cultured, Humans, Polysaccharides isolation & purification, Collagen metabolism, Fibroblasts drug effects, Hyaluronic Acid metabolism, Plant Extracts pharmacology, Polysaccharides pharmacology, Ulva chemistry
- Abstract
Ulvans from Ulva sp. were tested for their potential cosmetic properties on human dermal fibroblasts. The crude ulvans (ULVAN-01, 57kDa), extracted using a patented acid- and solvent-free process, were subjected to depolymerization using ion exchange resin to obtain a low molecular weight ulvan (ULVAN-DEP, 4kDa). The biochemical characterization and UHPLC-HRMS analyses of these extracted ulvans showed that they were of high purity and predominantly composed of a repeated ulvanobiouronic acid disaccharide. Fibroblast proliferation, as well as hyaluronan and collagen release were assessed, demonstrating that ULVAN-01 reduced fibroblast proliferation rate while ULVAN-DEP had no significant effect. Both ulvans were ineffective to induce collagen production but induced a significant increase in hyaluronan production, with a strong influence of the molecular weight. Thus, crude and depolymerized ulvans had different metabolic activities on dermal fibroblasts, which makes them promising to envisage further development in the skin care field., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2017
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16. Alteration of cathepsin D trafficking induced by hypoxia and extracellular acidification in MCF-7 breast cancer cells.
- Author
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Achour O, Ashraf Y, Bridiau N, Kacem M, Poupard N, Bordenave-Juchereau S, Sannier F, Lamerant-Fayel N, Kieda C, Liaudet-Coopman E, Piot JM, Maugard T, and Fruitier-Arnaudin I
- Subjects
- Cell Hypoxia genetics, Cell Hypoxia physiology, Cell Movement physiology, Female, Humans, Kinetics, MCF-7 Cells, Breast Neoplasms metabolism, Cathepsin D metabolism
- Abstract
The microenvironment that surrounds tumor cells is characterized by hypoxic conditions and extracellular acidity. These hostile conditions induce crucial changes in cell behavior and can promote the secretion of many soluble factors such as growth factors, cytokines and enzymes. The lysosomal aspartyl-endopeptidase cathepsin D (CD) is a marker of poor prognosis in breast cancer and is associated with a metastatic risk. In this study, the transport of CD was investigated in a model of breast cancer cells line (MCF-7) cultivated under hypoxia and acidification of media. CD secretion was assessed using Western blot analysis and protease activity was measured in conditioned culture media. We demonstrate that cultured MCF-7 cells secrete an active 52 kDa pCD precursor and report that under hypoxia there was an increased amount of pCD secreted. More surprisingly, extracellular acidification (pH 6 and 5.6) induced the secretion of the fully-mature and active (34 kDa + 14 kDa) double chain CD. Our findings reflect the fact that chemical anomalies influence the secretion path of CD in a breast cancer cell model, resulting in altered trafficking of the mature form. This important result may provide new arguments in favor of the role of extracellular CD in the degradation of the matrix proteins that constitute the breast tumor microenvironment., (Copyright © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.)
- Published
- 2016
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17. Anti-heparanase activity of ultra-low-molecular-weight heparin produced by physicochemical depolymerization.
- Author
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Achour O, Poupard N, Bridiau N, Bordenave Juchereau S, Sannier F, Piot JM, Fruitier Arnaudin I, and Maugard T
- Subjects
- Catalysis, Hydrogen Peroxide chemistry, Hydrolysis, Molecular Weight, Ultrasonic Waves, Anticoagulants chemistry, Glucuronidase chemistry, Heparin chemistry
- Abstract
Heparanase is an endo-β-D-glucuronidase that plays an important role in cancer progression, in particular during tumor angiogenesis and metastasis. Inhibiting this enzyme is considered as one of the most promising approaches in cancer therapy. Heparin is a complex glycoaminoglycan known as a strong inhibitor of heparanase. It is primarily used in clinical practice for its anticoagulant activities, which may not be compatible with its use as anti-angiogenic agent. In this study, we described the production of ultra-low-molecular-weight heparins (ULMWH) by a physicochemical method that consists in a hydrogen peroxide-catalyzed radical hydrolysis assisted by ultrasonic waves. We assessed the structural characteristics, anticoagulant and anti-heparanase activities of the obtained heparin derivatives and compared them with three commercial low-molecular-weight heparins (LMWH), glycol-split non-anticoagulant heparins and heparins produced by enzymatic methods. ULMWH generated by the physicochemical method were characterized by high anti-heparanase and moderate anticoagulant activities. These heparin derivatives might be potential candidates for cancer therapy when a compromise is needed between anti-heparanase and anticoagulant activities., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2016
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18. Characterization of the colistin (polymyxin E1 and E2) biosynthetic gene cluster.
- Author
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Tambadou F, Caradec T, Gagez AL, Bonnet A, Sopéna V, Bridiau N, Thiéry V, Didelot S, Barthélémy C, and Chevrot R
- Subjects
- Anti-Bacterial Agents isolation & purification, Anti-Bacterial Agents pharmacology, Base Sequence, Colistin isolation & purification, Colistin pharmacology, DNA, Bacterial analysis, DNA, Bacterial genetics, Microbial Sensitivity Tests, Multigene Family genetics, Paenibacillus genetics, Peptide Synthases genetics, Sequence Analysis, DNA, Tandem Mass Spectrometry, Anti-Bacterial Agents biosynthesis, Colistin biosynthesis, Paenibacillus metabolism, Peptide Synthases metabolism, Pseudomonas aeruginosa drug effects
- Abstract
Colistin is a mixture of polymyxin E1 and E2, bactericidal pentacationic lipopeptides used to treat infections caused by Gram-negative pathogens such as Pseudomonas aeruginosa and Klebsiella pneumoniae. Industrial production of colistin is obtained by a fermentation process of the natural producer Paenibacillus polymyxa var colistinus. NonRibosomal peptide synthetases (NRPS) coding the biosynthesis of polymyxins A, B and P have been recently described, rendering thereof the improvement of their production possible. However, the colistin biosynthesis pathway was not published so far. In this study, a Paenibacillus alvei has been identified by biochemical (Api 50 CH system) and molecular (16S rDNA sequencing) methods. Its culture supernatant displayed inhibitory activity against Gram-negative bacteria (P. aeruginosa, K. pneumoniae, Salmonella spp.). Two polymyxins, E1 and E2, were recovered from the supernatant and were characterized by high resolution LC-MS. A genomic library (960 clones) was constructed to identify the gene cluster responsible for biosynthesis of polymyxins. Selection of the clones harbouring the sequences of interest was obtained by a simple PCR-based screening. We used primers targeting NRPS sequences leading to the incorporation of amino acids present in polymyxins E. The sequences from three clones of interest were assembled on 50.4 kb. Thus, five open reading frames corresponding to a new NRPS gene cluster of 41 kb were identified. In silico, analyses revealed the presence of three NRPS implicated in the biosynthesis of polymyxins E. This work provides insightful information on colistin biosynthesis and might contribute to future drug developments in this group of antibiotics.
- Published
- 2015
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19. Dextranase immobilization on epoxy CIM(®) disk for the production of isomaltooligosaccharides from dextran.
- Author
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Bertrand E, Pierre G, Delattre C, Gardarin C, Bridiau N, Maugard T, Štrancar A, and Michaud P
- Subjects
- Dextranase chemistry, Enzymes, Immobilized chemistry, Equipment Design, Hydrolysis, Industrial Microbiology instrumentation, Dextranase metabolism, Dextrans metabolism, Enzymes, Immobilized metabolism, Epoxy Compounds chemistry, Methylmethacrylates chemistry, Oligosaccharides metabolism, Penicillium enzymology
- Abstract
Endodextranase D8144 from Penicillium sp. (EC 3.2.1.2.) was immobilized on an epoxy-activated monolithic Convective Interaction Media (CIM(®)) disk in order to produce isomaltooligosaccharides (IMOS) from Dextran T40 in a continuous IMmobilized Enzymes Reactor (IMER). Enzymatic parameters and structure of IMOS were studied for free and immobilized enzymes. The immobilization efficiency of endodextranase D8144 was about 15.9% (w/w) and the real specific activity was close to 6.5 U mg enz(-1). The Km values (4.8 ± 0.2 g L(-1)) for free and immobilized enzymes were the same, showing the absence of diffusional limitation. Moreover, specific patterns of DPs (Degrees of Polymerization) distributions were observed during the enzymatic hydrolysis by HPAEC-PAD (High Pressure Anion Exchange Chromatography-Pulsed Amperometric Detection). Thus, sought-after sizes of IMOS (DPs 8-10) were generated all over the hydrolysis. Finally, the results showed the high stability of this IMER since a relative enzymatic activity about 78% was measured after 5400 volumes column., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2014
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20. Measuring angiotensin-I converting enzyme inhibitory activity by micro plate assays: comparison using marine cryptides and tentative threshold determinations with captopril and losartan.
- Author
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Ben Henda Y, Labidi A, Arnaudin I, Bridiau N, Delatouche R, Maugard T, Piot JM, Sannier F, Thiéry V, and Bordenave-Juchereau S
- Subjects
- Enzyme Assays instrumentation, Angiotensin-Converting Enzyme Inhibitors analysis, Captopril analysis, Enzyme Assays methods, Losartan analysis, Peptide Fragments analysis, Peptidyl-Dipeptidase A analysis
- Abstract
To determine the angiotensin-I converting enzyme (ACE) inhibitory activity of marine cryptides, different methods were tested. ACE inhibition was measured using two synthetic substrates, (N-[3-(2-furyl) acryloyl]-Phe-Gly-Gly (FAPGG) and N-hippuryl-His-Leu hydrate salt (HHL)), and a natural one, angiotensin-I. The IC50 value (defined as the concentration of inhibitory molecule needed to inhibit 50% of the ACE activity) of the reference synthetic inhibitor captopril was in the nanomolar range (1.79-15.1 nM) when synthetic substrates were used, whereas it exhibited IC50 of micromolar range (16.71 μM) with angiotensin-I. We chose losartan, an antagonist of angiotensin-II receptor as negative control for the ACE inhibition. Losartan was also able to inhibit ACE whatever the substrate tested, with IC50 of micromolar range (17.13-146 μM). We defined this value as a limit above which molecules are not showing in vitro ACE inhibitory activity. Val-Trp (VW), Val-Tyr (VY), Lys-Tyr (KY), Lys-Trp (KW), Ile-Tyr (IY), Ala-Pro (AP), Val-Ile-Tyr (VIY), Leu-Lys-Pro (LKP), Gly-Pro-Leu (GPL), Ala-Lys-Lys (AKK), and Val-Ala-Pro (VAP) were tested as inhibitors of ACE with synthetic and natural substrates. IC50 displayed were substrate-dependent. With FAPGG as substrate, IW, VAP, KY, IY, AP, AKK, and VIY show IC50 values over the IC50 value of losartan and should not be considered as inhibitors of ACE. VY, VW, KW, and LKP exhibited IC50 value lower than the IC50 value of losartan for all substrates tested and were thus considered as good candidates for effectively decreasing hypertension. It appears that the comparison of IC50 is not consistent when IC50 values are obtained with different substrates and different methods. In vitro ACE inhibitory activity assays should always include various ACE substrates and references such as captopril and a negative control to obtain data reliable to discriminate ACE inhibitory peptides.
- Published
- 2013
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21. Cathepsin D activity and selectivity in the acidic conditions of a tumor microenvironment: Utilization in the development of a novel Cathepsin D substrate for simultaneous cancer diagnosis and therapy.
- Author
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Achour O, Bridiau N, Kacem M, Delatouche R, Bordenave-Juchereau S, Sannier F, Thiéry V, Piot JM, Maugard T, and Arnaudin I
- Subjects
- Catalysis, Cathepsin D chemistry, Cathepsin D genetics, Gene Expression Regulation, Neoplastic, Humans, Neoplasms pathology, Neoplasms therapy, Substrate Specificity, Tumor Microenvironment genetics, Cathepsin D metabolism, Neoplasms diagnosis, Tumor Microenvironment immunology
- Abstract
Pro-Cathepsin D (pCD) is an aspartyl endopeptidase which is over expressed in many cancers. This over expression generally led to its secretion into the extracellular culture medium of cancer cells. Moreover, pCD can auto activate and cleave its substrates at an acidic pH compatible with that found in tumor microenvironments (TME). Thus, exploiting these two pathological characteristics of TME offers the opportunity to develop new protease-activated vector on the basis of their specific substrate structures. The aim of this study was to validate new pCD substrates in the extracellular pH conditions of TME. As a first step, we investigated the effect of pH on the catalytic activity and selectivity of mature Cathepsin D (CD). It was found that the increase in the pH of the media led to a decrease in the reaction rate. However, the specificity of mature CD was not affected by a variation in pH. In the second step, the effect of the substrate structure was studied. We demonstrated that the substrate structure had a significant effect on the catalytic activity of CD. In fact, some modifications in peptide structure induced a change in the catalytic behavior that involved a substrate activation phenomenon. We suggest that this activation may be related to the amphiphilic nature of the modified peptide that may induce an interfacial activation mechanism. Finally, pCD, which is the major form found in the extracellular culture medium of cancer cells, was used. We demonstrated that the proform of CD cleave the modified peptide 5 at pH 6.5 with the same cleavage selectivity obtained with the mature form of the protease. These data provide a better understanding of CD behavior in tumor microenvironment conditions and this knowledge can be used to develop more specific tools for diagnosis and drug delivery., (Copyright © 2013. Published by Elsevier Masson SAS.)
- Published
- 2013
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22. Antiproliferative activity of Cyanophora paradoxa pigments in melanoma, breast and lung cancer cells.
- Author
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Baudelet PH, Gagez AL, Bérard JB, Juin C, Bridiau N, Kaas R, Thiéry V, Cadoret JP, and Picot L
- Subjects
- Antineoplastic Agents chemistry, Antineoplastic Agents pharmacology, Apoptosis drug effects, Cell Line, Tumor, Cryptoxanthins, Cyanophora metabolism, Female, Humans, MCF-7 Cells, Pigments, Biological chemistry, Skin Neoplasms, Xanthophylls chemistry, Xanthophylls pharmacology, Zeaxanthins, Melanoma, Cutaneous Malignant, Breast Neoplasms drug therapy, Cell Proliferation drug effects, Cyanophora chemistry, Lung Neoplasms drug therapy, Melanoma drug therapy, Pigments, Biological pharmacology
- Abstract
The glaucophyte Cyanophora paradoxa (Cp) was chemically investigated to identify pigments efficiently inhibiting malignant melanoma, mammary carcinoma and lung adenocarcinoma cells growth. Cp water and ethanol extracts significantly inhibited the growth of the three cancer cell lines in vitro, at 100 µg · mL(-1). Flash chromatography of the Cp ethanol extract, devoid of c-phycocyanin and allophycocyanin, enabled the collection of eight fractions, four of which strongly inhibited cancer cells growth at 100 µg · mL(-1). Particularly, two fractions inhibited more than 90% of the melanoma cells growth, one inducing apoptosis in the three cancer cells lines. The detailed analysis of Cp pigment composition resulted in the discrimination of 17 molecules, ten of which were unequivocally identified by high resolution mass spectrometry. Pheophorbide a, β-cryptoxanthin and zeaxanthin were the three main pigments or derivatives responsible for the strong cytotoxicity of Cp fractions in cancer cells. These data point to Cyanophora paradoxa as a new microalgal source to purify potent anticancer pigments, and demonstrate for the first time the strong antiproliferative activity of zeaxanthin and β-cryptoxanthin in melanoma cells.
- Published
- 2013
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23. Ultrasonic-assisted preparation of a low molecular weight heparin (LMWH) with anticoagulant activity.
- Author
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Achour O, Bridiau N, Godhbani A, Le Joubioux F, Bordenave Juchereau S, Sannier F, Piot JM, Fruitier Arnaudin I, and Maugard T
- Subjects
- Animals, Catalysis drug effects, Chemical Fractionation, Chromatography, Gel, Factor Xa metabolism, Heparin, Low-Molecular-Weight pharmacology, Hydrolysis drug effects, Molecular Weight, Polymerization drug effects, Prothrombin metabolism, Sus scrofa, Anticoagulants chemical synthesis, Heparin, Low-Molecular-Weight chemical synthesis, Ultrasonics
- Abstract
Low molecular weight heparin (LMWH) is currently used as an anticoagulant agent and constitutes an alternative to unfractionated heparin, which is the cause of serious adverse drug reaction such as heparin-induced thrombocytopenia (HIT). Commercially available LMWH is produced by enzymatic depolymerization that is costly or by chemical methods that are generally carried out under conditions that could imply side reactions that reduce final product efficiency and yields. In this work, we present the use of a physicochemical method for the production of LMWH. This method consists in the use of hydrogen peroxide-catalyzed radical hydrolysis assisted by ultrasonic waves. LMWH that are produced using this physicochemical method have an average molecular weight and anticoagulant properties (Anti-Xa and Anti-IIa) that are comparable to some of commercial LMWH that are currently used. Ultrasonic-assisted radical depolymerization of heparin leads to products with a remarkably low polydispersity index. Moreover, in comparison to other LMWH such as those produced by enzymatic β-elimination, this physicochemical depolymerization of heparin induces fewer oligosaccharides with less than five monosaccharide units. This contributes to the better preservation of the ATIII pentasaccharide binding sequence, which results in a high Anti-Xa/Anti-IIa ratio (1.86). However, LMWH obtained using this physicochemical method have a lower degree of sulfation than other LMWH, which seems to be the cause of a lower Anti-Xa and Anti-IIa activity (143.62±5.42 and 77.07±4.4, respectively)., (Copyright © 2013 Elsevier Ltd. All rights reserved.)
- Published
- 2013
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24. A comparative study of the regioselectivity of the β-galactosidases from Kluyveromyces lactis and Bacillus circulans in the enzymatic synthesis of N-Acetyl-lactosamine in aqueous media.
- Author
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Bridiau N and Maugard T
- Subjects
- Acetylglucosamine metabolism, Amino Sugars metabolism, Substrate Specificity, Water, Amino Sugars biosynthesis, Bacillus enzymology, Kluyveromyces enzymology, beta-Galactosidase metabolism
- Abstract
The enzymatic synthesis of N-acetyl-lactosamine (LacNAc) was studied in aqueous media with high substrate concentrations using the transgalactosylation of N-acetyl-D-glucosamine (GlcNAc), starting from lactose as a galactosyl donor. The efficiency and regioselectivity of the β-galactosidases from Kluyveromyces lactis (KlβGal) and Bacillus circulans (BcβGal) were compared. The reaction was optimized by varying the experimental conditions (pH, catalytic activity concentration, and mass concentration ratio of the substrates), which enhanced the synthesis yields with both enzymes and especially with BcβGal. BcβGal catalyzed the formation of the maximal LacNAc concentration obtained (101 mM or 39 g L(-1), corresponding to a yield of 11% on the basis of GlcNAc conversion), after 5 h at pH 6.5 and for a substrate mass concentration ratio of 1. This enzyme also gave an optimal synthesis yield of about 17.5%. No change in regioselectivity was observed when using KlβGal, whereas the regioselectivity of BcβGal proved to be subject to variations, the 1-4 and 1-6 linkages being favored under kinetic and thermodynamic control conditions, respectively. Finally, it was demonstrated that the N-acetyl-allolactosamine synthesized during the GlcNAc transgalactosylation catalyzed by BcβGal was a thermodynamic product and did not result from a chemical and/or enzymatic isomerization of LacNAc., (Copyright © 2011 American Institute of Chemical Engineers (AIChE).)
- Published
- 2011
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25. The effects of organic solvents on the efficiency and regioselectivity of N-acetyl-lactosamine synthesis, using the β-galactosidase from Bacillus circulans in hydro-organic media.
- Author
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Bridiau N, Issaoui N, and Maugard T
- Subjects
- Acetylglucosamine chemistry, Acetylglucosamine metabolism, Chromatography, High Pressure Liquid, Enzyme Stability, Magnetic Resonance Spectroscopy, Stereoisomerism, Thermodynamics, Amino Sugars chemistry, Amino Sugars genetics, Amino Sugars metabolism, Bacillus enzymology, Solvents chemistry, beta-Galactosidase metabolism
- Abstract
The enzymatic synthesis of N-acetyl-lactosamine (LacNAc) by the transgalactosylation of N-acetyl-D-glucosamine (GlcNAc), catalyzed by the β-galactosidase from Bacillus circulans (BcβGal), was studied in hydro-organic media, starting from o-nitrophenyl-β-D-galactopyranoside (oNPG) as a galactosyl donor. Thermal stability and synthesis activity of BcβGal were shown to depend on the organic solvent polarity, characterized by its Log P value. BcβGal was thus most stable in 10% (v/v) t-BuOH, an organic solvent found to have a stabilizing and/or weakly denaturing property, which was confirmed for high t-BuOH concentrations. In the same manner, the optimal synthesis yield increased as the Log P value of the organic solvent increased. The best results were obtained for reactions carried out in 10% (v/v) pyridine or 2-methyl-2-butanol, which gave 47% GlcNAc transgalactosylation yield based on starting oNPG, of which 23% (11 mM; 4.3 g/L) consisted in LacNAc synthesis. Furthermore, it was also established that both the GlcNAc transgalactosylation yield and the enzyme regioselectivity depended on the percentage of organic solvent used, the optimal percentage varying from 10 to 40% (v/v), depending on the solvent. This phenomenon was found to correlate mainly with the thermodynamic activity of water (a(w)) in the aqueous organic solvent mixture, which was found to be optimal when close to 0.96, whatever the organic solvent used. Finally, this study highlighted the fact that the regioselectivity of BcβGal for 1-4 linkage formation could be advantageously managed by controlling the a(w) parameter., (© 2010 American Institute of Chemical Engineers)
- Published
- 2010
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26. Beta-galactosidase catalyzed selective galactosylation of aromatic compounds.
- Author
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Bridiau N, Taboubi S, Marzouki N, Legoy MD, and Maugard T
- Subjects
- Adsorption, Catalysis, Chlorphenesin chemistry, Chlorphenesin metabolism, Glycosylation, Guaifenesin chemistry, Guaifenesin metabolism, Hydrocarbons, Aromatic chemistry, Molecular Structure, Hydrocarbons, Aromatic metabolism, Kluyveromyces enzymology, beta-Galactosidase metabolism
- Abstract
A new approach to galacto-oligosaccharides and galacto-conjugates synthesis performed by the beta-galactosidase from Kluyveromyces lactis is reported. The enzymatic galactosylation of eight kinds of adsorbed aromatic primary alcohols, in particular the two drugs guaifenesin and chlorphenesin, gave the corresponding beta-D-galacto-pyranosides in yields ranging between approximately 10% and 96%. For the first time, we have showed that the adsorption of acceptor substrates onto solid supports such as silica gel influences the yield and the selectivity of galacto-conjugates synthesis. In particular, we observed that adsorption of acceptor favored the synthesis of digalactosylated compounds.
- Published
- 2006
- Full Text
- View/download PDF
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