Journal
CARBOHYDRATE POLYMERS
Volume 129, Issue -, Pages 194-200Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carbpol.2015.04.068
Keywords
Hyaluronan; Recombinant leech hyaluronidase; Molecular mass distribution; Oligosaccharides; Polydispersity; Large-scale enzymatic production
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Funding
- Key Technologies R & D Program of Jiangsu Province, China [BE2014607]
- National High Technology Research and Development Program of China (863 Program) [2012AA022005]
- Program for Changjiang Scholars and Innovative Research Team in University [IRT1135]
- National Natural Science Foundation of China [31200020]
- 111 Project
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High-molecular-mass hyaluronan (HA) was controllably depolymerized in pure aqueous solution with recombinant leech hyaluronidase (HAase). The HAase concentration per unit HA and hydrolysis time played important roles in molecular mass distribution. By modulating the concentrations of HAase and controlling the hydrolysis time, any molar-mass-defined HA oligomers could be efficiently and specifically produced on a large scale (40 g/L), such as HA oligosaccharides with weight-average molar mass of 4000, 10,000, and 30,000 Da and end hydrolysates containing only HA6 and HA4. High performance liquid chromatography-size exclusion chromatography, polyacrylamide gel electrophoresis, capillary zone electrophoresis, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry confirmed low polydispersity of the produced molar-mass-defined HA oligosaccharides. Therefore, large-scale production of defined HA oligosaccharides with narrow molecular mass distribution will significantly promote progress in related research and its potential applications. (C) 2015 Elsevier Ltd. All rights reserved.
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