Optimization of Bacillus subtilis natto Growth Parameters in Glycerol-Based Medium for MK-7 (Vitamin K) Production in Biofilm Reactors

Abstract. Menaquinone -7 (MK-7) is the key form of vitamin K used as a dietary supplement and its production revolves around Bacillus subtilis natto. Current fermentation strategies, which suggest static fermentations without aeration and agitation, can be problematic for large scale MK-7 production due to biofilm formation. The use of biofilm reactors, therefore, is proposed in the present study, which could utilize both agitation and aeration without interrupting MK-7 secretion. In this study, biofilm reactors were constructed using the selected PCS and B. subtilis natto strain for MK-7 production. Using response surface methodology (RSM), optimum growth parameters including temperature, pH, and agitation were determined in a glycerol-based medium. Results were presented in a statistical model (R²=0.90), leading to optimum growth conditions of temperature (35°C), agitation (200 rpm) and pH (6.58). Model predicted MK-7 concentration was validated and MK-7 concentration of 12.09 mg/L was produced in the biofilm reactor. The obtained concentration was 58% higher as compared to the suspended-cell culture. The results of this study will provide a critical step towards improved industrial scale production of MK-7.