Structural characterization of an exopolysaccharide produced by two strains of Lacticaseibacilluscasei

Exopolysaccharides (EPSs) were isolated and purified from Lacticaseibacillus casei strains type V and RW-3703M grown under various fermentation conditions (carbon source, incubation temperature, and duration). Identical ¹H NMR spectra were obtained in all cases. The molar mass determined by size-exclusion chromatography coupled with multi-angle light scattering was different for the two strains and in different culture media. The primary structure was elucidated using chemical and spectroscopic techniques. Monosaccharide and absolute configuration analyses gave the following composition: d-Glc, 1; d-Gal, 2; l-Rha, 2; d-GlcNAc, 1. Methylation analysis indicated the presence of 4-linked Glc, terminal and 6-linked Gal, terminal and 3-linked Rha, and 3,4,6-linked GlcNAc. On the basis of one- and two-dimensional ¹H and ¹³C NMR data, the structure of the EPS was consistent with the following hexasaccharide repeating unit: {4)[Rhap(α1-3)][Galp(α1-6)]GlcpNAc(β1-6)Galp(α1-3)Rhap(β1-4)Glcp(β1-}_n. Complete ¹H and ¹³C NMR assignments are reported.