Anaerobic autotrophic bacteria Clostridium ljungdahlii is capable of producing ethanol and acetate from sugars or synthesis gas in aqueous nutrient mediums. The objective of this study was to evaluate optimum cultural conditions on cell growing such as initial pH, yeast extract, fructose fed and temperature in batch culture for the production of ethanol and acetate. A series of experiments were conducted under strictly anaerobic conditions with 1atm at N2 headspace pressure in serum vials. The results showed that higher cell concentration of C. ljungdahlii cell was achieved to 0.57 g/L (OD660~1.0) at interval of initial pH of 6.8-7.0. And then the ethanol and acetate production reached to 0.92 g/L and 2.1 g/L after 72 hours. It was found that cell concentration and ethanol production were increased when 2 g/L of yeast extract and 5 g/L of fructose added in batch culture. The results confirmed that cell growth of C. ljungdahlii can be promoted by initial pH to 7.0 in anaerobic fermentation.