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Updated: Feb 2, 2026

Single-cell Analysis of Bacillus subtilis Biofilms Using Fluorescence Microscopy and Flow Cytometry
Published on: February 15, 2012
Enhanced Vitamin K (Menaquinone-7) Production by Bacillus subtilis natto in Biofilm Reactors by Optimization of
Ehsan Mahdinia1, Ali Demirci1, Aydin Berenjian2
1Department of Agricultural and Biological Engineering, The Pennsylvania State University, University Park, PA, 16802, United States.
Background:
Benefits of vitamin K have been reported by many studies recently, due to its ability to reduce the risk of cardiovascular diseases and its potential benefits against osteoporosis. Specifically, menaquinone-7 (MK-7), being the most potent form of vitamin K, has definitely received most of the attention. Currently, solid or static liquid fermentation strategies are utilized for industrial production of MK-7 by Bacillus strains. However, these strategies face fundamental operational and scale-up issues as well as intense pellicle and biofilm formations which is problematic in static liquid fermentation, due to heat and mass transfer inefficiencies they create.
Objective:
The purpose of this study was to demonstrate that biofilm reactors will overcome the issues associated with suspended cell reactors when using Bacillus strains to produce MK-7. The expectation is that the use of biofilm reactors will result in a significant increase in the production of MK-7.
Method:
Vitamin K production by Bacillus subtilis natto when grown in a biofilm reactor was evaluated at various concentrations of the three major nutrients, glucose, yeast extract and casein. The data was analyzed using response surface methodology (RSM).
Results:
The maximum concentration of MK-7 in the biofilm reactors was 20.5±0.5 mg/L, which was a 344 % increase when compared to the amount produced in suspended-cell reactors containing the same optimum media composition.
Conclusion:
These results demonstrate the potential of utilizing biofilm reactors for MK-7 production on an industrial scale.
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