Effects of medium components in a glycerol-based medium on vitamin K (menaquinone-7) production by Bacillus subtilis natto in biofilm reactors
Menaquinone-7 (MK-7) as the most important form of Vitamin K has been reported to have miraculous benefits such as preventing cardiovascular diseases and osteoporosis along with antitumor effects. Therefore, there have been numerous studies in the past decades to improve MK-7 production via microbial fermentation. Unfortunately, both solid and liquid state fermentation strategies that are utilized for MK-7 production, face fundamental operational and scale-up issues as well as intense heat and mass transfer problems during fermentation. In this regard, biofilm reactors seem to be a practical solution to overcome these issues and enhance the production in agitated liquid fermentation. Therefore, this study was undertaken to utilize biofilm reactors in investigating and optimizing different media components in a glycerol-based medium. Using response surface methodology, the effects of glycerol, yeast extract, and soytone were studied in the fermentation medium on MK-7 production in biofilm reactor. With a composition of 48.2 g/L of glycerol, 8.1 g/L of yeast extracts, 13.6 g/L of soytone and 0.06 g/L of K2HPO4, MK-7 concentrations could reach 14.7 ± 1.4 mg/L in biofilm reactors, which was 57% higher compared to the MK-7 concentration achieved in suspended-cell reactors under similar conditions, while glycerol was depleted by the end of the fifth day in biofilm reactors, but glycerol was never depleted in suspended-cell reactors. Evidently, biofilm reactors present a reliable strategy to address the operational issues that occur during MK-7 biosynthesis on an industrial scale production.
KeywordsMK-7 Menaquinone-7 Vitamin K Biofilm reactor Bacillus subtilis RSM optimization
This work was supported by the USDA National Institute of Food and Agriculture Federal Appropriations under Project PEN04561 and Accession number 1002249. The authors thank the Statistical Consulting Center at The Pennsylvania State University for their support in providing useful consultation for data processing.
- 2.Widhalm JR, Ducluzeau A-L, Buller NE, Elowsky CG, Olsen LJ, Basset GJC (2012) Phylloquinone (vitamin K1) biosynthesis in plants: two peroxisomal thioesterases of Lactobacillales origin hydrolyze 1,4-dihydroxy-2-naphthoyl-CoA. Plant J 71:205–215. https://doi.org/10.1111/j.1365-313X.2012.04972.x CrossRefGoogle Scholar
- 4.Binkley SB, Maccorquodale DW, Thayer A, Doisy EA (1939) The isolation of vitamin K1. J Biol Chem 130:219–234Google Scholar
- 7.Howard LM, Payne AG (2006) Health benefits of vitamin K2: a revolutionary natural treatment for heart disease and bone loss, 1st edn. Basic Health Publications, Inc., Laguna BeachGoogle Scholar
- 8.Gast GCM, de Roos NM, Sluijs I, Bots ML, Beulens JWJ, Geleijnse JM, Witteman JC, Grobbee DE, Peeters PHM, van der Schouw YT (2009) A high menaquinone intake reduces the incidence of coronary heart disease. Nutr Metab Cardiovasc Dis 19:504–510. https://doi.org/10.1016/j.numecd.2008.10.004 CrossRefGoogle Scholar
- 12.Walther B, Chollet M (2017) Menaquinones, bacteria, and foods: vitamin K2 in the diet. In: Vitamin K2—vital for health and wellbeing. IntechOpen, London, UK, pp 63–82. https://doi.org/10.5772/63712
- 14.Berenjian A, Mahanama R, Talbot A, Biffin R, Regtop H, Kavanagh J (2011) The effect of amino-acids and glycerol addition on MK-7 production. In: Proc. world congr. eng. comput. sci. II, pp 19–21Google Scholar
- 15.Goodman SR, Marrs BL, Narconis RJ, Olson RE (1976) Isolation and description of a menaquinone mutant from Bacillus licheniformis. J Bacteriol 125:282–289Google Scholar
- 19.Ikeda H, Doi Y (1990) A vitamin-K2-binding factor secreted from Bacillus subtilis. Eur J Biochem 192:219–224. https://doi.org/10.1111/j.1432-1033.1990.tb19218.x CrossRefGoogle Scholar
- 22.Demirci A, Pongtharangkul T, Pometto IIIAL (2007) Applications of biofilm reactors for production of value-added products by microbial fermentation. Blackwell Publishing and The Institude of Food Technologists, IowaGoogle Scholar
- 25.Ho KG, Pometto ALI, Hinz PN, Dickson JS, Demirci A (1997) Ingredient selection for plastic composite supports for l-(1)-lactic acid biofilm fermentation by Lactobacillus casei subsp. rhamnosus. Appl Environ Microbiol 63:2516–2523Google Scholar
- 33.Mahdinia E, Demirci A, Berenjian A (2018) Utilization of glucose-based medium and optimization of Bacillus subtilis natto growth parameters for vitamin K (menaquinone-7) production in bio fi lm reactors. Biocatal Agric Biotechnol 13:219–224. https://doi.org/10.1016/j.bcab.2017.12.009 CrossRefGoogle Scholar