The effect of dissolved oxygen(DO) on microbial transglutaminase(mTG) production by Streptoverticillium morbaraense was studied in on-line computer controlled fermentation system. In order to control dissolved oxygen during fermentation, the agitation speed and aeration rate of 2.5 L fermenter ranged from 260 to 360 rpm and 0.3 to 3.9 L/min, respectively. The maximum microbial transglutaminase production was obtained at controlled 20% of dissolved oxygen among the various dissolved oxygen controlled batch cultures tested. The production of microbial transglutaminase at controlled 20% of dissolved oxygen was about 2.12 U/mL which was 1.1 times higher than that obtained in batch culture without control of dissolved oxygen. Also, the highest microbial transglutaminase production was obtained in fed-batch cultures in which dissolved oxygen was controlled at 20%, and it was improved almost 1.3 times in comparison with that without control of dissolved oxygen. Maximal dry cell weight and microbial transglutaminase production were 13.2 g/L and 2.6 U/mL, respectively. Finally, it was also found that fed-batch fermentation at controlled 20% of dissolved oxygen showed a good performance for the microbial transglutaminase production by on-line computer controlled fermentation system which may be generally applicable to other microbial cultures.