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 plastic composite support (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 (R 2 = 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 (7.67 mg/L). The results of this study will provide a critical step towards improved industrial scale production of MK-7.
Similar content being viewed by others
References
Dam H (1935) The antihaemorrhagic vitamin of the chick: occurrence and chemical nature. Nature 135:652–653
Binkley SB, Maccorquodale DW, Thayer A, Doisy EA (1939) The isolation of vitamin K1. J Biol Chem 130:219–234
Widhalm JR, Ducluzeau A-L, Buller NE, Elowsky CG, Olsen LJ, Basset GJC (2012) Phylloquinone (vitamin K(1)) biosynthesis in plants: two peroxisomal thioesterases of Lactobacillales origin hydrolyze 1,4-dihydroxy-2-naphthoyl-CoA. Plant J 71:205–215. doi:10.1111/j.1365-313X.2012.04972.x
Davidson RT, Foley AL, Engelke JA, Suttie JW (1998) Conversion of dietary phylloquinone to tissue menaquinone-4 in rats is not dependent on gut bacteria. J Nutr 128:220–223
Mahdinia E, Demirci A, Berenjian A (2017) Production and application of menaquinone-7 (vitamin K2): a new perspective. World J Microbiol Biotechnol 33:2. doi:10.1007/s11274-016-2169-2
Bentley R, Meganathan R (1982) Biosynthesis of vitamin K (menaquinone) in bacteria. Microbiol Rev 46:241–280
Berenjian A, Mahanama R, Kavanagh J, Dehghani F (2015) Vitamin K series: current status and future prospects. Crit Rev Biotechnol 35:199–208. doi:10.3109/07388551.2013.832142
Schurgers LJ, Teunissen KJF, Hamulyák K, Knapen MHJ, Vik H, Vermeer C (2007) Vitamin K-containing dietary supplements: comparison of synthetic vitamin k1 and natto-derived menaquinone-7. Blood 109:3279–3283. doi:10.1182/blood-2006-08-040709
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 Beach
Shi J, Zhou S, Kang L, Ling H, Chen J, Duan L (2017) Evaluation of the antitumor effects of vitamin K 2 (menaquinone-7) nanoemulsions modified with sialic acid–cholesterol conjugate. Drug Deliv Transl Res 2:1–11. doi:10.1007/s13346-017-0424-1
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. doi:10.1016/j.numecd.2008.10.004
Geleijnse JM, Vermeer C, Grobbee DE, Schurgers LJ, Knapen MHJ, van der Meer IM, Hofman A, Witteman JCM (2004) Dietary intake of menaquinone is associated with a reduced risk of coronary heart disease: the Rotterdam Study. J Nutr 134:3100–3105
Yamaguchi M (2006) Regulatory mechanism of food factors in bone metabolism and prevention of osteoporosis. Yakugaku Zasshi 126:1117–1137. doi:10.1248/yakushi.126.1117
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 congress on engineering and computer science, vol II, 19–21 October, San Francisco
Goodman SR, Marrs BL, Narconis RJ, Olson RE (1976) Isolation and description of a menaquinone mutant from Bacillus licheniformis. J Bacteriol 125:282–289
Wu W-J, Ahn B-Y (2011) Improved menaquinone (vitamin k2) production in Cheonggukjang by optimization of the fermentation conditions. Food Sci Biotechnol 20:1585–1591. doi:10.1007/s10068-011-0219-y
Singh R, Puri A, Panda BP (2015) Development of menaquinone-7 enriched nutraceutical: inside into medium engineering and process modeling. J Food Sci Technol 52:5212–5219. doi:10.1007/s13197-014-1600-7
Pandey A (2003) Solid-state fermentation. Biochem Eng J 13:81–84. doi:10.1016/S1369-703X(02)00121-3
Ikeda H, Doi Y (1990) A vitamin-K2-binding factor secreted from Bacillus subtilis. Eur J Biochem 192:219–224. doi:10.1111/j.1432-1033.1990.tb19218.x
Mahdinia E, Demirci A (2017) Strain and plastic composite support (PCS) selection for vitamin K (menaquinone-7) production in biofilm reactors. Bioprocess Biosyst Eng 40:1507–1517. doi:10.1007/s00449-017-1807-x
Berenjian A, Chan NL-C, Mahanama R, Talbot A, Regtop H, Kavanagh J, Dehghani F (2013) Effect of biofilm formation by Bacillus subtilis natto on menaquinone-7 biosynthesis. Mol Biotechnol 54:371–378. doi:10.1007/s12033-012-9576-x
Ercan D, Demirci A (2013) Current and future trends for biofilm reactors for fermentation processes. Crit Rev Biotechnol 8551:1–14. doi:10.3109/07388551.2013.793170
Demirci A, Pongtharangkul T, Pometto III AL (2007) Applications of biofilm reactors for production of value-added products by microbial fermentation. Blackwell Publishing and The Institude of Food Technologists, Iowa
Kuchma SL, O’Toole GA (2000) Surface-induced and biofilm-induced changes in gene expression. Curr Opin Biotechnol 11:429–433. doi:10.1016/S0958-1669(00)00123-3
Ercan D, Demirci A (2013) Production of human lysozyme in biofilm reactor and optimization of growth parameters of Kluyveromyces lactis K7. Appl Microbiol Biotechnol 97:6211–6221. doi:10.1007/s00253-013-4944-4
Berenjian A, Mahanama R, Talbot A, Biffin R, Regtop H, Valtchev P, Kavanagh J, Dehghani F (2011) Efficient media for high menaquinone-7 production: response surface methodology approach. N Biotechnol 28:665–672. doi:10.1016/j.nbt.2011.07.007
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 63:2516–2523
Ercan D, Demirci A (2014) Enhanced human lysozyme production in biofilm reactor by Kluyveromyces lactis K7. Biochem Eng J 92:2–8. doi:10.1016/j.bej.2014.04.013
Izmirlioglu G, Demirci A (2016) Ethanol production in biofilm reactors from potato waste hydrolysate and optimization of growth parameters for Saccharomyces cerevisiae. Fuel 181:643–651. doi:10.1016/j.fuel.2016.05.047
Rahimi M, Zhu L, Kowalski KL, Zhu X, Gorski CA, Hickner MA, Logan BE (2017) Improved electrical power production of thermally regenerative batteries using a poly(phenylene oxide) based anion exchange membrane. J Power Sources 342:956–963. doi:10.1016/j.jpowsour.2017.01.003
Rahimi M, Schoener Z, Zhu X, Zhang F, Gorski CA, Logan BE (2017) Removal of copper from water using a thermally regenerative electrodeposition battery. J Hazard Mater 322:551–556. doi:10.1016/j.jhazmat.2016.10.022
Sato T, Yamada Y, Ohtani Y, Mitsui N, Murasawa H, Araki S (2001) Production of menaquinone (vitamin K2)-7 by Bacillus subtilis. J Biosci Bioeng 91:16–20. doi:10.1016/S1389-1723(01)80104-3
Lindgren V, Rutberg L (1974) Glycerol metabolism in Bacillus subtilis: gene-enzyme relationships. J Bacteriol 119:431–442
Sato T, Yamada Y, Ohtani Y, Mitsui N, Murasawa H, Araki S (2001) Efficient production of menaquinone (vitamin K2) by a menadione-resistant mutant of Bacillus subtilis. J Ind Microbiol Biotechnol 26:115–120. doi:10.1038/sj.jim.7000089
Acknowledgements
This study was funded in-part by the Pennsylvania Agricultural Experiment Station. The authors thank the Statistical Consulting Center at The Pennsylvania State University for their support in providing useful consultation for data processing.
Author information
Authors and Affiliations
Corresponding author
Additional information
Chemical compound studied in this article: Menaquinone-7 (PubChem CID: 5287554); glycerol (PubChem CID: 753); glucose (PubChem CID: 79025); dipotassium hydrogen phosphate (PubChem CID: 24450); sulfuric acid (PubChem CID: 1118); sodium hydroxide (PubChem CID: 14798); n-hexane (PubChem CID: 8058); 2-propanol (PubChem CID: 3776); methanol (PubChem CID: 887).
Rights and permissions
About this article
Cite this article
Mahdinia, E., Demirci, A. & Berenjian, A. Optimization of Bacillus subtilis natto growth parameters in glycerol-based medium for vitamin K (Menaquinone-7) production in biofilm reactors. Bioprocess Biosyst Eng 41, 195–204 (2018). https://doi.org/10.1007/s00449-017-1857-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-017-1857-0