Polyglucuronic Acid Research Articles

Polyglucuronic acids prepared from α-(1 → 3)-glucan by TEMPO-catalytic oxidation

2,2,6,6-Tetramethylpiperidine-1-oxyl radical (TEMPO)-catalytic oxidation was applied to a water-insoluble α-(1 → 3)-glucan in water at pH 10 and room temperature (∼24 °C), with solid NaOCl·5H2O as the primary oxidant. Oxidation with NaOCl at 15 mmol/g gave a water-soluble TEMPO-oxidized product at a mass recovery ratio of 97 %. The carboxy content of the TEMPO-oxidized product was 5.3 mmol/g, which corresponds to a degree of C6-oxidation (DO) of 93 %. A new water-soluble α-(1 → 3)-polyglucuronic acid with a nearly homogeneous chemical structure was therefore quantitatively obtained. X-ray diffraction and solid-state 13C NMR spectroscopic analyses showed that the original α-(1 → 3)-glucan and its TEMPO-oxidized product with a carboxy content of 5.3 mmol/g had crystalline structures, whereas the oxidized products with DOs of 50 % and 66 % had almost disordered structures. The carboxy groups in the oxidized products were regioselectively methyl esterified with trimethylsilyl diazomethane, and analyzed by using size-exclusion chromatography with multi-angle laser-light scattering and refractive index detections. The results show that the original α-(1 → 3)-glucan and its oxidized products with DOs of 50 %, 66 %, and 93 % had weight-average degrees of polymerization of 671, 288, 54, and 45, respectively. Substantial depolymerization of the α-(1 → 3)-glucan molecules therefore occurred during catalytic oxidation, irrespective of the oxidation pH.

Water-Soluble Biomass Resist Materials Based on Polyglucuronic Acid for Eco-Friendly Photolithography

This study presents the development of photolithography employing biomass-based resist materials derived from polyglucuronic acid. Traditional resist materials require coating and developing procedures involving organic solvents, whereas our approach enables the use of water-based spin-coating and developing processes. The water-soluble biomass resist material, derived from polyglucuronic acid, exhibited exceptional photosensitivity at an exposure wavelength of 365 nm and a dose of approximately 90 mJ/cm2. We successfully patterned the microstructures, creating 3 µm lines and 6 µm holes. This organic solvent-free coating process underscores its applicability in scenarios such as in the microfabrication on plastic substrates with limited organic solvent tolerance and surface-patterning biomaterials containing cells and culture components.

Fabrication of cell-laden microbeads and microcapsules composed of bacterial polyglucuronic acid

In the past decades, the microencapsulation of mammalian cells into microparticles has been extensively studied for various in vitro and in vivo applications. The aim of this study was to demonstrate the viability of bacterial polyglucuronic acid (PGU), an exopolysaccharide derived from bacteria and composed of glucuronic acid units, as an effective material for cell microencapsulation. Using the method of dropping an aqueous solution of PGU-containing cells into a Ca2+-loaded solution, we produced spherical PGU microbeads with >93 % viability of the encapsulated human hepatoma HepG2 cells. Hollow-core microcapsules were formed via polyelectrolyte complex layer formation of PGU and poly-l-lysine, after which Ca2+, a cross-linker of PGU, was chelated, and this was accomplished by sequential immersion of microbeads in aqueous solutions of poly-l-lysine and sodium citrate. The encapsulated HepG2 cells proliferated and formed cell aggregates within the microparticles over a 14-day culture, with significantly larger aggregates forming within the microcapsules. Our results provide evidence for the viability of PGU for cell microencapsulation for the first time, thereby contributing to advancements in tissue engineering.

Description of the Wild Strain Rhizobium rosettiformans DSM26376, Reclassified under Peteryoungia rosettiformans comb.nov., for Producing Glucuronan.

Glucuronan is a polysaccharide composed of β-(1,4)-linked d-glucuronic acids having intrinsic properties and biological activities recoverable in many fields of application. Currently, the description of Sinorhyzobium meliloti M5N1CS mutant bacterial strain as the sole source of glucuronan makes it relevant to the exploration of new microorganisms producing glucuronan. In this study, the Peteryoungia rosettifformans strain (Rhizobia), was identified as a wild producer of an exopolysaccharide (RhrBR46) related to glucuronan. Structural and biochemical features, using colorimetric assays, Fourier infrared spectroscopy, nuclear magnetic resonance, high pressure size exclusion chromatography coupled to multi-angle light laser scattering, and enzymatic assays allowed the characterization of a polyglucuronic acid, having a molecular mass (Mw¯) of 1.85 × 105 Da, and being partially O-acetylated at C-2 and/or C-3 positions. The concentration of Mg2+ ions in the cultivation medium has been shown to impact the structure of RhrBR46, by reducing drastically its Mw¯ (73%) and increasing its DA (10%). Comparative structural analyses between RhrBR46 and the glucuronan from Sinorhyzobium meliloti M5N1CS strain revealed differences in terms of molecular weight, degree of acetylation (DA), and the distribution of acetylation pattern. These structural divergences of RhrBR46 might contribute to singular properties or biological activities of RhrBR46, offering new perspectives of application.

Multifunctional Enzyme with Endoglucanase and Alginase/Glucuronan Lyase Activities from Bacterium Cellulophaga lytica.

Cellulophaga lytica is a Gram-negative aerobic bacterium in the genome of which there are many genes encoding polysaccharide degrading enzymes. One of the enzymes named ClGP contains a glycoside hydrolase domain from the GH5 family and a polysaccharide lyase domain from the PL31 family. The enzyme also contains the TAT signaling peptide and the TIGR04183 domain that indicates extracellular nature of the enzyme. Phylogenetic analysis has shown that the enzymes most closely related to ClGP and containing all four domains (TAT, GH5, PL31, TIGR04183) are widespread among bacterial species belonging to the Flavobacteriaceae family. ClGP produced by the recombinant strain of E. coli was purified and characterized. ClGP exhibited activity of endoglucanase (EC 3.2.1.4) and catalyzed hydrolysis of β-D-glucan, carboxymethyl cellulose sodium salt (CMC-Na), and amorphous cellulose, but failed to hydrolyze microcrystalline cellulose and xylan. Products of CMC hydrolysis were cellobiose and cellotriose, whereas β-D-glucan was hydrolyzed to glucose, cellobiose, cellotetraose, and cellopentaose. ClGP was more active against the poly-β-D-mannuronate blocks than against the poly-α-L-glucuronate blocks of alginic acid. This indicates that the enzyme is a polyM lyase (EC 4.2.2.3). ClGP was active against polyglucuronic acid, so it displayed a glucuronan lyase (EC 4.2.2.14) activity. The enzyme had a neutral pH-optimum, was stable in the pH range 6.0-8.0, and displayed moderate thermal stability. ClGP effectively saccharified two species of brown algae, Saccharina latissima and Laminaria digitata, that suggests its potential for use in the production of biofuel from macroalgae.

Bacterial Polyglucuronic Acid/Alginate/Carbon Nanofibers Hydrogel Nanocomposite as a Potential Scaffold for Bone Tissue Engineering.

3D nanocomposite scaffolds have attracted significant attention in bone tissue engineering applications. In the current study, we fabricated a 3D nanocomposite scaffold based on a bacterial polyglucuronic acid (PGU) and sodium alginate (Alg) composite with carbon nanofibers (CNFs) as the bone tissue engineering scaffold. The CNFs were obtained from electrospun polyacrylonitrile nanofibers through heat treatment. The fabricated CNFs were incorporated into a PGU/Alg polymeric solution, which was physically cross-linked using CaCl2 solution. The fabricated nanocomposites were characterized to evaluate the internal structure, porosity, swelling kinetics, hemocompatibility, and cytocompatibility. The characterizations indicated that the nanocomposites have a porous structure with interconnected pores architecture, proper water absorption, and retention characteristics. The in vitro studies revealed that the nanocomposites were hemocompatible with negligible hemolysis induction. The cell viability assessment showed that the nanocomposites were biocompatible and supported bone cell growth. These results indicated that the fabricated bacterial PGU/Alg/CNFs hydrogel nanocomposite exhibited appropriate properties and can be considered a new biomaterial for bone tissue engineering scaffolds.

Development of phenol-grafted polyglucuronic acid and its application to extrusion-based bioprinting inks

In this present work, we developed a phenol grafted polyglucuronic acid (PGU) and investigated the usefulness in tissue engineering field by using this derivative as a bioink component allowing gelation in extrusion-based 3D bioprinting. The PGU derivative was obtained by conjugating with tyramine, and the aqueous solution of the derivative was curable through a horseradish peroxidase (HRP)-catalyzed reaction. From 2.0 w/v% solution of the derivative containing 5 U/mL HRP, hydrogel constructs were successfully obtained with a good shape fidelity to blueprints. Mouse fibroblasts and human hepatoma cells enclosed in the printed constructs showed about 95% viability the day after printing and survived for 11 days of study without a remarkable decrease in viability. These results demonstrate the great potential of the PGU derivative in tissue engineering field especially as an ink component of extrusion-based 3D bioprinting.

Discovery of a Novel Glucuronan Lyase System in Trichoderma parareesei.

Glucuronan lyases (EC 4.2.2.14) catalyze depolymerization of linear β-(1,4)-polyglucuronic acid (glucuronan). Only a few glucuronan lyases have been characterized until now, most of them originating from bacteria. Here we report the discovery, recombinant production, and functional characterization of the full complement of six glucuronan specific polysaccharide lyases in the necrotic mycoparasite Trichoderma parareesei. The enzymes belong to four different polysaccharide lyase families and have different reaction optima and glucuronan degradation profiles. Four of them showed endo-lytic action and two, TpPL8A and TpPL38A, displayed exo-lytic action. Nuclear magnetic resonance revealed that the monomeric end product from TpPL8A and TpPL38A underwent spontaneous rearrangements to tautomeric forms. Proteomic analysis of the secretomes from T. parareesei growing on pure glucuronan and lyophilized A. bisporus fruiting bodies, respectively, showed secretion of five of the glucuronan lyases and high-performance anion-exchange chromatography with pulsed amperometric detection analysis confirmed the presence of glucuronic acid in the A. bisporus fruiting bodies. By systematic genome annotation of more than 100 fungal genomes and subsequent phylogenetic analysis of the putative glucuronan lyases, we show that glucuronan lyases occur in several ecological and taxonomic groups in the fungal kingdom. Our findings suggest that a diverse repertoire of glucuronan lyases is a common trait among Hypocreales species with mycoparasitic and entomopathogenic lifestyles. IMPORTANCE This paper reports the discovery of a set of six complementary glucuronan lyase enzymes in the mycoparasite Trichoderma parareseei. Apart from the novelty of the discovery of these enzymes in T. parareesei, the key importance of the study is the finding that the majority of these lyases are induced when T. parareesei is inoculated on Basidiomycete cell walls that contain glucuronan. The study also reveals putative glucuronan lyase encoding genes in a wealth of other fungi that furthermore points at fungal cell wall glucuronan being a target C-source for many types of fungi. In a technical context, the findings may lead to controlled production of glucuronan oligomers for advanced pharmaceutical applications and pave the way for development of new fungal biocontrol agents.

Long-Term Fate of Magnetic Particles in Mice: A Comprehensive Study.

Safe application of nanoparticles in medicine requires full understanding of their pharmacokinetics including catabolism in the organism. However, information about nanoparticle degradation is still scanty due to difficulty of long-term measurements by invasive techniques. Here, we describe a magnetic spectral approach for in vivo monitoring of magnetic particle (MP) degradation. The method noninvasiveness has allowed performing of a broad comprehensive study of the 1-year fate of 17 types of iron oxide particles. We show a long-lasting influence of five parameters on the MP degradation half-life: dose, hydrodynamic size, ζ-potential, surface coating, and internal architecture. We observed a slowdown in MP biotransformation with an increase of the injected dose and faster degradation of the particles of a small hydrodynamic size. A comparison of six types of 100 nm particles coated by different hydrophilic polymer shells has shown that the slowest (t1/2 = 38 ± 6 days) and the fastest (t1/2 = 15 ± 4 days) degradations were achieved with a polyethylene glycol and polyglucuronic acid coatings, respectively. The most significant influence on the MP degradation was due to the internal architecture of the particles as the coverage of magnetic cores with a solid 39 nm polystyrene layer slowed down the half-life of the core-shell MPs from 48 days to more than 1 year. The revealed deeper insights into the particle degradation in vivo may facilitate rational design of nano- and microparticles with predictable long-term fate in vivo.

Identification and characterization of cellouronate (β-1,4-linked polyglucuronic acid) lyase from the scallop Mizuhopecten yessoensis

The semisynthetic polysaccharide cellouronate is a β-1,4-linked polyglucuronic acid prepared from regenerated cellulose by chemical oxidation. Here, we isolated a novel enzyme, MyAly, as a cellouronate lyase from a scallop Mizuhopecten yessoensis. Its optimum temperature, pH, and NaCl concentration for cellouronate degradation were determined to be 30 °C, 6.9, and 200–500 mM, respectively. MyAly endolytically degraded cellouronate into unsaturated di-, tri-, and tetrasaccharides with kcat of 31.1 s−1. MyAly also showed an alginate-degradation activity with a kcat value of 0.58 s−1. However, there was no significant difference in Km values between cellouronate and alginate. MyAly consisted of 280 amino acids and shared 36.5–44.1 % identity with known marine gastropod alginate lyases belonging to the polysaccharide lyase family 14. This is the first study to identify and characterize a cellouronate-degrading lyase from a marine organism, providing a better understanding of the biodegradability of the industrially important polysaccharide, cellouronate, in marine environments.

Marine Bacteria, A Source for Alginolytic Enzyme to Disrupt Pseudomonas aeruginosa Biofilms.

Pseudomonas aeruginosa biofilms are typically associated with the chronic lung infection of cystic fibrosis (CF) patients and represent a major challenge for treatment. This opportunistic bacterial pathogen secretes alginate, a polysaccharide that is one of the main components of its biofilm. Targeting this major biofilm component has emerged as a tempting therapeutic strategy for tackling biofilm-associated bacterial infections. The enormous potential in genetic diversity of the marine microbial community make it a valuable resource for mining activities responsible for a broad range of metabolic processes, including the alginolytic activity responsible for degrading alginate. A collection of 36 bacterial isolates were purified from marine water based on their alginolytic activity. These isolates were identified based on their 16S rRNA gene sequences. Pseudoalteromonas sp. 1400 showed the highest alginolytic activity and was further confirmed to produce the enzyme alginate lyase. The purified alginate lyase (AlyP1400) produced by Pseudoalteromonas sp. 1400 showed a band of 23 KDa on a protein electrophoresis gel and exhibited a bifunctional lyase activity for both poly-mannuronic acid and poly-glucuronic acid degradation. A tryptic digestion of this gel band analyzed by liquid chromatography-tandem mass spectrometry confirmed high similarity to the alginate lyases in polysaccharide lyase family 18. The purified alginate lyase showed a maximum relative activity at 30 °C at a slightly acidic condition. It decreased the sodium alginate viscosity by over 90% and reduced the P. aeruginosa (strain PA14) biofilms by 69% after 24 h of incubation. The combined activity of AlyP1400 with carbenicillin or ciprofloxacin reduced the P. aeruginosa biofilm thickness, biovolume and surface area in a flow cell system. The present data revealed that AlyP1400 combined with conventional antibiotics helped to disrupt the biofilms produced by P. aeruginosa and can be used as a promising combinational therapeutic strategy.

An engineered polysaccharide lyase to combat harmful algal blooms

A growing global population and industrialization have come at the cost of induced climate change and pollution of natural resources, resulting in formation of toxic algal blooms in fresh water sources. In the US alone, these blooms cost an estimated $1.5 billion dollars each year to remediate. Current methods to combat such blooms such as copper sulfate treatment are expensive, ineffective, and environmentally toxic, motivating development of biochemical algaecides as green alternatives. Our previous research led to identification of mutant polysaccharide lyase (PL) enzyme H208F derived from Stenotrophomonas maltophilia Smlt2602 (WT), which displays increased activity on the polysaccharide polyglucuronic acid. Polyglucuronic acid is a major component of the algal cell wall. This study focused on applying the two enzymes to a common bloom-forming, toxin-producing algae, Microcystis aeruginosa. We hypothesized the 2 enzymes would kill algae through disrupting the cell wall, with H208F displaying a higher killing efficacy over WT. A fluorescence assay in conjunction with live/dead staining was used to quantify killing of algae treated with each enzyme. SEM analysis was used to observe cell wall morphology in order to verify the killing mechanism. The mutant H208F was confirmed as having potential application as a biochemical, enzymatic algaecide with a 17% greater algicidal efficacy than the WT. SEM images revealed deformities in the algal cell wall, confirming the purported mechanism.

Oxidation pattern of curdlan with TEMPO-mediated system

In this study, the TEMPO-mediated (TEMPO/NaBr/NaClO) oxidation pattern of curdlan was investigated through comprehensively structural analysis of the corresponding oxidized products. During the structural analysis, infrared spectroscopy (IR), nuclear magnetic resonance (NMR) spectroscopy, gel permeation chromatography tandem multiple angle laser scattering (GPC-MALS) and ultra-high performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UHPLC-Q/TOF-MS) were applied. As a result, the homogenous β1-3 polyglucuronic acids (MW, 49.8, 29.8 and 7.0 kDa) were obtained with proper amount of oxidant (5.36 mmol NaClO) at various temperatures (4, 25, 50 °C), respectively. Compared to the oxidation of 1–4 linked glucan (starch and cellulose) with TEMPO-mediated system at same reaction conditions, higher degree of specific oxidation and less degradation were observed in that of 1–3 linked curdlan. The glycosylation at position 3 could stabilize the sugar ring, which inactivates the non-specific oxidation related hydroxyl groups on the sugar ring. Thus, the TEMPO-mediated system has higher selectivity to oxidize the primary hydroxyl groups of 1–3 linked curdlan and form polyglucuronic acid than those observed in the oxidation of starch and cellulose. In addition, same as those observed in previous work about starch, higher the temperature was used in the oxidation with TEMPO system, higher the activity of oxidant (NaClO solution) was, more non-specific oxidation occurred, and more the degradation were observed.

Synthesis of amphoteric curdlan derivatives for delivery of therapeutic nucleic acids

Cationic polymers are powerful carriers for intracellular delivery of therapeutic nucleic acids. However, the positively charged macromolecules have considerable cytotoxicity and often induce irreversible damages to the cells and tissues, which greatly negate the clinical application of such materials as drug delivery system. Herein, we report the synthesis of novel amphoteric polymers based on curdlan, and the evaluation of their cytotoxicity as well as the nucleic acid delivery efficiency. β-(1,3)-polyglucuronic acid, a TEMPO-oxidized derivative of curdlan, was chemically modified by conjugation of tetraethylenepentamine. The resulting amphoteric polymers, denoted tetraethylenepentamine-curdlan (TEPAC) polymers have the degree of substitution (DS) ranging from 25% to 48%. The result of MTT assay indicated that TEPAC polymers have negligible cytotoxicity on HeLa cells and A549 cells. The novel amphoteric polymers efficiently bound with plasmid DNA and delivered pcDNA-eGFP plasmid to 293T cells and induced expression of GFP 48h after the transfection. Moreover, TEPAC polymers delivered siRNA to HeLa cells and HepG2 cells in high efficiency, and induced significant RNAi for the expression of an endogenous gene. Collectively, our data demonstrate that the novel curdlan-based amphoteric polymers are biocompatible and may provide a highly efficient system for the delivery of therapeutic nucleic acids.

Antioxidant activities of a polyglucuronic acid sodium salt obtained from TEMPO-mediated oxidation of xanthan.

A xanthouronic acid sodium salt called xanthouronan was produced from xanthan by regioselective oxidation with NaOCl/NaBr using 2,2,6,6-tetramethylpiperidine-1-oxy radical (TEMPO) as catalyst. The efficiency of the one pot TEMPO-mediated oxidation was confirmed by HPAEC-PAD, (13)C NMR, and FT-IR. The oxidation degree was close to 98% and the mass yield of this new polyglucuronic acid was higher than 90% (w/w). The macromolecular characterization of xanthouronan using SEC-MALLS showed a molecular size reduced by a third due to the oxidation treatment and the degree of polymerization (DP) of the xanthouronan form was about 665. The evaluation of the enzymatic degradation of this C-6 carboxylated xanthan by various polysaccharide hydrolases and one polysaccharide lyase showed its high resistant to biodegradation. The antioxidant activity of xanthouronan was also tested by using the 2,2'-diphenyl-1-picrylhydrazyle (DPPH) and hydroxyl radical procedures. At 1 g/L, xanthouronan presented 75% of the ascorbic acid antioxidant activity.

Water soluble exo-polysaccharide from Syncephalastrum racemosum, a strong inducer of plant defence reactions

This study examines the production, characterization and bioactivity on plant cell cultured in vitro of exopolysaccharides (EPS) from Syncephalastrum racemosum CBS 443.59. Firstly, the influence of the fungus culture condition in shake flasks (pH, temperature and different carbon and nitrogen sources) on EPS and biomass production was evaluated. In order to enhance EPS production, a new protocol based on two-stage pH fermentation in a 3L stirred fermentor was developed. Under this condition, EPS production increased by 3.55 times, compared to a constant pH process, reaching a maximal EPS concentration of 2.62g/L. Structurally, the EPS contains a polyglucuronic acid backbone, linked essentially with mannose and fucose units and some galactose and glucose units. The bioactivity of EPS as inducer of defence reactions in plant suspension-cultured cells was also studied. Our results show, for first time, that EPS from S. racemosum CBS 443.59 induces, depending on the concentration, PAL activation and H2O2 synthesis in Arabidopsis thaliana cell suspensions.

Facile synthesis of a new fluorogenic metal scavenging interpolymeric diamide based on cellulose and alginic acids

A microwave assisted synthesis of a water soluble fluorogenic interpolymeric diamide has been described involving alginic acid and polyglucuronic acid (PGA) amide of ethylenediamine (EDA), through a monoamide of PGA and EDA, in good yields (>80wt% in each step). PGA was prepared by TEMPO (2,2,6,6-tetramethyl piperidine-1-oxyl radical) mediated oxidation of cellulose of the halophytic plant Salicornia brachiata. The amides were characterized by spectral analyses. The fluorescence emission of the PGA amide was 7-fold greater than that of the interpolymeric diamide. PGA monoamide exhibited superior heavy metal ions [Pb(II) and Hg(II)] uptake properties to the diamide, the former showing optimum adsorptions of ions 398.8 and 282.8mg/g, respectively. These materials may be of utility as potential sensors harnessing their fluorogenic and metal scavenging properties.

Structural characteristics and antioxidant activities of different families of 4-acetamido-TEMPO-oxidised curdlan

Regioselective oxidation was applied to commercial curdlan for the preparation of its water-soluble derivatives with improved antioxidant activities, using a 4-acetamido-2,2, 6,6-tetramethylpiperidine-1-oxyl radical/NaClO/NaClO2 system at pH 4.8 and 40°C. The structural features, molecular properties, and chain conformations of the oxidised curdlans were determined using Fourier transform (FT) infrared and FT Raman spectroscopy, carbon nuclear magnetic resonance, X-ray diffraction, and size-exclusion chromatography with multi-angle laser-light scattering analyses. The C6 primary hydroxyls of curdlan were successfully oxidised into carboxylate groups with less depolymerization, and no aldehyde groups were formed during oxidation. The crystalline region of curdlan was destroyed after oxidation. The oxidised curdlans formed random coils in aqueous solution, and the chain became more flexible and expanded with increasing carboxylate contents from 2.07mmol/g to 4.87mmol/g. The high polyglucuronic acid derivative (Cur-24) showed the best antioxidant activity in TEAC and FRAP assays, thus it could be explored as novel potential antioxidants for dietary and therapeutic applications.

Supramolecular structure and anomer-selective formation of polyglucuronic acid

Polyglucuronic acids provide a biocompatible matrix for industries ranging from pharmaceuticals to agriculture. Here we provide a method for selectively producing α-polyglucuronic acid, a simple alternative to the oxidation of cellulose or amylose with 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO) or isolation from bacterial cultures. Atomic force micrographs show the pH-dependent formation and persistence of polyglucuronic acid fibrils, while NMR studies demonstrate the selectivity of the glycosidic linkage. By controlling pH, oligoglucuronate chains are created, with both NMR and AFM data showing the preservation of this linkage in basic medium after formation under acid. Clear evidence for the formation of the C–O–C glycosidic bond comes from peaks in the 13C NMR spectra, revealing a linkage through the anomeric carbon atom and also a linkage with a ring carbon atom.

Synthesis of new glycosaminoglycans-like families by regioselective oxidation followed by sulphation of glucoglucuronan from Rhizobium sp. T1

Glycosaminoglycan-like polysaccharides were prepared from Rhizobium sp. T1 polysaccharide using the TEMPO (nitroxyl radical 2,2,6,6-tetramethylpiperidine-1-oxyl radical) mediated oxidation. The structure of this new polyglucuronic acid sodium salt was analyzed by 13C NMR spectra and HPAEC–PAD chromatography. Therefore, new polysaccharide containing only glucuronic acid monomers in both β-(1,3) and β-(1,4) linkage was obtained by the complete TEMPO-mediated oxidation of C6 primary hydroxyl groups of glucose of glucoglucuronan from Rhizobium sp. T1. Sulphation of this β-(1,3),β-(1,4)-polyglucuronic acid sodium salt was carried out using SO3/DMF reagent. These results suggested a new synthetic route using both TEMPO-mediated oxidation and sulphation of polysaccharides from Rhizobium sp. in developing glycosaminoglycans mimic to enhance the profitability of its low-cost production and processing industries. This novel carbohydrate derivative might find use as cheaper surrogates of glycosaminoglycans in the cosmetics and pharmaceutical fields.