Taxonomic group: human / Chordata
(Phylum: Chordata)
NCBI PubMed ID: 31222385Publication DOI: 10.1007/s00253-019-09969-8Journal NLM ID: 8406612Publisher: Springer
Correspondence: Odile Francesca Restaino <odilefrancesca.restaino
unicampania.it>; Donatella Cimini <donatella.cimini
unicampania.it>
Institutions: Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, University of Campania 'Luigi Vanvitelli', Via De Crecchio 7, 80138, Naples, Italy
Heparin and chondroitin sulfate are used as anti-thrombic and anti-osteoarthritis drugs, respectively, but their pharmacological actions depend on their structural characteristics such as their sulfation grade and their molecular weight. In the last years, new fermentation-based biotechnological approaches have tried to obtain heparin and chondroitin sulfate starting from the heparosan and chondroitin-like capsular polysaccharides produced by Escherichia coli K5 and K4. The study of the microbial capsular polysaccharide molecular weight is critical to obtain nature-like or structural tailor cut glycosaminoglycan homologues. However, so far, it has been scarcely investigated. In this paper, for the first time, a new protocol was set up to determine the molecular weights of the capsular polysaccharides of three wild-type and three engineered E. coli K5 and K4 strains. The protocol includes a small-scale downstream train to purify the intact polysaccharides, directly from the fermentation broth supernatants, by using ultrafiltration membranes and anion exchange chromatography, and it couples size exclusion chromatography analyses with triple detector array. In the purification high recovery (> 85.0%) and the removal of the main contaminant, the lipopolysaccharide, were obtained. The averaged molecular weights of the wild-type capsular polysaccharides ranged from 51.3 to 90.9 kDa, while the engineered strains produced polysaccharides with higher molecular weights, ranging from 68.4 to 130.6 kDa, but with similar polydispersity values between 1.1 and 1.5.
Escherichia coli, capsular polysaccharide, experimental, heparin, molecular weight, Chondrotin sulfate, SEC-TDA
Structure type: polymer chemical repeating unit ; 14000-70000
Location inside paper: p.6771
Trivial name: chondroitin sulfate (CS)
Contained glycoepitopes: IEDB_115136,IEDB_130648,IEDB_137473,IEDB_140630,IEDB_153510,IEDB_423153
Methods: HPCE, fermentation, anion exchange chromatography, HPGPC, ultrafiltration, SEC-TDA
Comments, role: repeating units could be mono- or di-sulfated.
Related record ID(s): 1095, 2259, 2260
NCBI Taxonomy refs (TaxIDs): 9606Reference(s) to other database(s): GTC:G43702JT
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There is only one chemically distinct structure:
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