The optimization of fermentation medium is important for synthetic biological secondary metabolite productions. The effect of rotation speed, inoculum amount, and medium supplements on the cell growth and LycogenTM secretion of photobacterium Rhodobacter sphaeroides WL-APD911 was evaluated. The results reveal that a higher rotational speed exhibit a higher cell density, and the increasing in the amount of inoculum amount show a slight augment on the growth of R. sphaeroides WL-APD911.In the case of nitrogen sources adding, LycogenTM production was achieved with a 0.5mM l-lysine supplementation. Moreover, the attention of Tween 80 presented a tremendous increase in the secondary metabolite. Response surface methodology (RSM) exhibited the optimization of medium supplements for LycogenTM invention is accomplished at molasses concentration of 10 g/L, yeast extract concentration of 40 g/L, 0.3% Tween 80 and NaCl concentration of 5 g/L, respectively. Further, the batch fermentation is carried out in both 5 L and 20 L fermentors to study the scale-up process factors to be adopted. At a 20 L fermentor, LycogenTM yields under the optimal culture condition are over 2 times than in the shake flask. The present results provide the LycogenTM optimal culture mediums, scale-up procedures and efficient extractions from R. sphaeroides WL-APD911.