Terrific Broth(TB) medium was proved to be the optimum medium for β-1,3-1,4-glucanase fermentation using recombinant Escherichia coli.Several factors including 10 carbon sources and 9 nitrogen sources were studied in order to improve the productivity of β-1,3-1,4-glucanase by recombinant E.coli BL21DE3(pET-28a(+)-bgl).And glycerin,yeast extract and tryptone were finally selected as the best carbon source and nitrogen sources,respectively.Response surface methodology(RSM),fractional factorial design(FFD),steepest ascent path,and central composite design(CCD) were used to determine the optimum concentrations of each component in TB medium.Finally the concentrations were determined as follows: yeast extract 20 g/L,tryptone 12.5 g/L,glycerin 14.1 mL/L,KH2PO4 2.17 g/L and K2HPO4 2.74 g/L.And activity of β-1,3-1,4-glucanase achieved 2978 U/mL,which was 1.78 times higher than that from original medium.These conclusions will provide theoretical basis for the pilot scale fermentation and further utilization of the β-1,3-1,4-glucanase in industry.