An exopolysaccharide (EPS) producer was isolated from milk sample by primary screening. It was identified as Klebseilla pneumoniae strain 27F on basis of biochemical studies and 16s rDNA gene based analysis. Semi–defined medium composition and other physical parameters viz. pH, temperature and time were varied for high exopolysaccharide production. The influence of variable carbon and nitrogen sources were investigated. Klebseilla  pneumoniae in  semi-defined medium(SDM) at pH 6.5 containing lactose (60g/l), NH₄Cl (17g/l), K₂HPO₄ (2.5g/l), MnSO₄ (0.1g/l) and MgSO₄ (0.05g/l)  under shaker conditions for 6 days at 30⁰C produced 4.45 g/l of EPS. The other growth parameter like absence of Na-acetate also showed remarkable 2.5 fold increase in EPS yield. HPTLC analysis of EPS from Klebseilla pneumoniae strain 27F showed presence of galactose and mannose.