A new extracellular polysaccharide (EPS-I) isolated and purified from Z222, a strain of Lactic acid bacteria has been investigated. Sugar composition analysis, methylation analysis and ^1H NMR and ^13C NMR spectroscopy reveal that the EPS-I is composed of a pentasaccharide repeating unit. The sequence of sugar residue was determined by using two-dlmensional NMR spectroscopy, including heteronudear multiple-bond correlation(HMBC) and nuclear overhauser effect spectroscopy (NOESY).