期刊
CARBOHYDRATE POLYMERS
卷 167, 期 -, 页码 123-128出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.carbpol.2017.03.026
关键词
Heparosan; Chondroitin; Recombinant E. coli; Enzymatic synthesis; Click chemistry
Conjugatable glycosaminoglycans hold promise for medical applications involving the vectorization of specific molecules. Here, we set out to produce bacterial chondroitin and heparosan from a conjugatable precursor using metabolically engineered Escherichia coli strains. The major barrier to this procedure was the glucuronylation of a lactosyl acceptor required for polymerization. To overcome this barrier, we designed E. coli strains expressing mouse beta-1,3-glucuronyl transferase and E. coli K4 chondroitin and K5 heparosan synthases. These engineered strains were cultivated at high density in presence of a lactosefuryl precursor. Enzymatic polymerization occurred on the lactosyl precursor resulting in small chains ranging from 15 to 30 kDa that accumulated in the cytoplasm. Furyl-terminated polysaccharides were produced at a gram-per-liter scale, a yield similar to that reported for conventional strains. Their efficient conjugation using a Diels-Alder cycloaddition reaction in aqueous and catalyst-free conditions was also confirmed using N-methylmaleimide as model dienophile. (C) 2017 Elsevier Ltd. All rights reserved.
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