期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 67, 期 24, 页码 6856-6866出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.9b01822
关键词
dextran; mutan; NMR spectroscopy; purification; HPAEC-PAD; oligosaccharides; enzymatic fingerprinting; fermentation
资金
- Carl-Zeiss Foundation
- German Federal Ministry of Economic Affairs and Energy [AiF19180N]
The water kefir organism Leuconostoc citreum TMW 2.1194 forms highly branched dextrans with O3- and O4- bound side chains. To obtain detailed information on the enzymatic synthesis of these polymers, the four glucansucrases encoded by Leuconostoc citreum TMW 2.1194 were cloned, heterologously expressed, and used for polysaccharide production. Molecular and macromolecular structure of the synthesized glucans were analyzed by methylation analysis, two-dimensional NMR spectroscopy, oligosaccharide analysis after partial hydrolysis, and asymmetric flow field-flow fractionation. It was demonstrated that two glucansucrases form insoluble glucans with variously branched dextran sections and varying portions of consecutive, 1,3-linked glucose units. In contrast, the other two glucansucrases synthesized O3- (Lc6255) and O4-branched (Lc1785) soluble dextrans. Analysis, isolation, and characterization of enzymatically liberated oligosaccharides showed that monomeric and elongated side chains are abundant in both polysaccharides. From the structures and size distributions it was concluded that Lc1785 is mainly responsible for synthesis of fermentatively produced soluble dextrans.
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