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Abstract: Agrobacterium sp. was studied for the production of curdlan by conventional one-factor-at-a-time technique and response surface methodology. Factors such as initial pH, urea concentration, sucrose concentration having the greatest influence on the curdlan production were identified. By using response surface methodology (RSM), the curdlan production by Agrobacterium sp. was increased significantly by 109%, from 2.4g/L to 5.02g/L when the strain was cultivated in the optimal medium developed by RSM as compared to conventional one-factor-at-a-time technique. The curdlan production rate of 0.84g/(Lh) was obtained when Agrobacterium sp. was cultivated in the optimal medium developed by RSM, which was the highest curdlan production rate reported to date. The infrared (IR) and NMR spectra, the thermogram of DSC and pattern of X-ray diffraction for the curdlan of the present study were almost identical to those of the authentic curdlan sample (from Alcaligenes faecalis; Sigma). The purified curdlan was a linear polysaccharide composed of exclusively β-(1,3)-glucosidic linkages with the molecular weight of 160,000Da by GPC. The crystalline melting point (T m), glass transition temperature (T g) and X-ray diffraction of the sample indicated low crystallinity in the structure. [Copyright &y& Elsevier]