Abstract
Menaquinone-7 (MK-7), a valuable member of the vitamin K2 series, is an essential nutrient for humans. It is used for treating coagulation disorders, and osteoporosis, promoting liver function recovery, and preventing cardiovascular diseases. In this study, to further improve the metabolic synthesis of MK-7 by the mutant strain, the effect of surfactants on the metabolic synthesis of MK-7 by the mutant strain Bacillus subtilis 168 KO-SinR (BS168 KO-SinR) was analyzed. The scanning electron microscopy and flow cytometry results showed that the addition of surfactants changed the permeability of the cell membrane of the mutant strain and the structural components of the biofilm. When 0.7% Tween-80 was added into the medium, the extracellular and intracellular synthesis of MK-7 reached 28.8 mg/L and 59.2 mg/L, respectively, increasing the total synthesis of MK-7 by 80.3%. Quantitative real-time PCR showed that the addition of surfactant significantly increased the expression level of MK-7 synthesis-related genes, and the electron microscopy results showed that the addition of surfactant changed the permeability of the cell membrane. The research results of this paper can serve as a reference for the industrial development of MK-7 prepared by fermentation.
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Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
References
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(6):665–672. https://doi.org/10.1016/j.nbt.2011.07.007
Berenjian A, Chan NL, 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(2):371–378. https://doi.org/10.1007/s12033-012-9576-x
Berenjian A, Mahanama R, Kavanagh J, Dehghani F (2015) Vitamin K series: current status and future prospects. Crit Rev Biotechnol 35(2):199–208. https://doi.org/10.3109/07388551.2013.832142
Branda SS, Vik S, Friedman L, Kolter R (2005) Biofilms: the matrix revisited. Trends Microbiol 13(1):20–26. https://doi.org/10.1016/j.tim.2004.11.006
Cui SX, Lv XQ, Wu YK, Li JH, Du GC, Ledesma-Amaro R, Liu L (2019) Engineering a bifunctional Phr60-Rap60-Spo0A quorum-sensing molecular switch for dynamic fine-tuning of menaquinone-7 synthesis in Bacillus subtilis. ACS Synth Biol 8(8):1826–1837. https://doi.org/10.1021/acssynbio.9b00140
Cui SX, Xia HZ, Chen TC, Gu Y, Lv XQ, Liu YF, Li JH, Du GC, Liu L (2020) Cell membrane and electron transfer engineering for improved synthesis of menaquinone-7 in Bacillus subtilis. iScience 23(3):100918. https://doi.org/10.1016/j.isci.2020.100918
Edgar RC (2010) Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26(19):2460–2461. https://doi.org/10.1093/bioinformatics/btq461
Fang X, Yang Q, Liu H, Wang P, Wang L, Zheng ZM, Zhao GH (2018) Effects of a combined processing technology involving ultrasound and surfactant on the metabolic synthesis of vitamin K2 by Flavobacterium sp. m1–14. Chem Eng Process 135:227–235. https://doi.org/10.1016/j.cep.2018.09.010
Fujimoto N, Kosaka T, Yamada M (2012) Menaquinone as well as ubiquinone as a crucial component in the Escherichia coli respiratory chain. Chem Biol 17:187–208. https://doi.org/10.5772/35809
Gast GC, de Roos NM, Sluijs I, Bots ML, Beulens JW, Geleijnse JM, Witteman JC, Grobbee DE, Peeters PH, van der Schouw YT (2009) A high menaquinone intake reduces the incidence of coronary heart disease. Nutr Metab Cardiovasc Dis 19(7):504–510. https://doi.org/10.1016/j.numecd.2008.10.004
Hewitt CJ, Nebe-Von-Caron G (2001) An industrial application of multiparameter flow cytometry: assessment of cell physiological state and its application to the study of microbial fermentations. Cytometry 44(3):179–187. https://doi.org/10.1002/1097-0320(20010701)44:3%3c179::aid-cyto1110%3e3.0.co;2-d
Holmberg C, Beijer L, Rutberg B, Rutberg L (1990) Glycerol catabolism in Bacillus subtilis: nucleotide sequence of the genes encoding glycerol kinase (glpK) and glycerol-3-phosphate dehydrogenase (glpD). J Gen Microbiol 136(12):2367–2375. https://doi.org/10.1099/00221287-136-12-2367
Hu XC, Liu WM, Luo MM, Ren LJ, Ji XJ, Huang H (2017) enhancing menaquinone-7 production by Bacillus natto R127 through the nutritional factors and surfactant. Appl Biochem Biotechnol 182(4):1630–1641. https://doi.org/10.1007/s12010-017-2423-6
Ikeda H, Doi Y (1990) A vitamin-K2-binding factor secreted from Bacillus subtilis. Eur J Biochem 192(1):219–224. https://doi.org/10.1111/j.1432-1033.1990.tb19218.x
Krämer M, Bongaerts J, Bovenberg R, Kremer S, Müller U, Orf S, Wubbolts M, Raeven L (2003) Metabolic engineering for microbial production of shikimic acid. Metab Eng 5(4):277–283. https://doi.org/10.1016/j.ymben.2003.09.001
Kurosu M, Begari E (2010) Vitamin K2 in electron transport system: are enzymes involved in vitamin K2 biosynthesis promising drug targets? Molecules 15(3):1531–1553. https://doi.org/10.3390/molecules15031531
Li LF, Liu Y, Jiang L, Ding S, Chen GG, Liang ZQ, Zeng W (2021) Effects of cell physiological structure on the fermentation broth viscosity during poly-γ-glutamic acid production by Bacillus subtilis GXA-28. Appl Biochem Biotechnol 193(1):271–280. https://doi.org/10.1007/s12010-020-03418-3
Lu PX, Wu AQ, Zhang S, Bai J, Guo T, Lin QL, Liu J (2021) Triton X-100 supplementation regulates growth and secondary metabolite biosynthesis during in-depth extractive fermentation of Monascus purpureus. J Biotechnol 341:137–145. https://doi.org/10.1016/j.jbiotec.2021.09.018
Luo MM, Ren LJ, Chen SL, Ji XJ, Huang H (2016) Effect of media components and morphology of Bacillus subtilis natto on menaquinone-7 synthesis in submerged fermentation. Biotechnol Bioproc E 21(6):777–786. https://doi.org/10.1007/s12257-016-0202-9
Ma Y, McClure DD, Somerville MV, Proschogo NW, Dehghani F, Kavanagh JM, Coleman NV (2019) Metabolic engineering of the MEP pathway in Bacillus subtilis for increased biosynthesis of menaquinone-7. ACS Synth Biol 8(7):1620–1630. https://doi.org/10.1021/acssynbio.9b00077
Mahdinia E, Demirci A, Berenjian A (2018) Optimization of Bacillus subtilis natto growth parameters in glycerol-based medium for vitamin K (Menaquinone-7) production in biofilm reactors. Bioprocess Biosyst Eng 41(2):195–204. https://doi.org/10.1007/s00449-017-1857-0
Miyake N, Hoshi K, Sano Y, Kikuchi K, Tadano K, Koshihara Y (2001) 1,25-Dihydroxyvitamin D3 promotes vitamin K2 metabolism in human osteoblasts. Osteoporos Int 12(8):680–687. https://doi.org/10.1007/s001980170068
O’Toole G, Kaplan HB, Kolter R (2000) Biofilm formation as microbial development. Annu Rev Microbiol 54:49–79. https://doi.org/10.1146/annurev.micro.54.1.49
Parsek MR, Greenberg EP (2005) Sociomicrobiology: the connections between quorum sensing and biofilms. Trends Microbiol 13(1):27–33. https://doi.org/10.1016/j.tim.2004.11.007
Pfaff MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucl Acids Res 29(9):e45. https://doi.org/10.1093/nar/29.9.e45
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(1):16–20. https://doi.org/10.1263/jbb.91.16
Shea MK, Holden RM (2012) Vitamin K status and vascular calcification: evidence from observational and clinical studies. Adv Nutr 3(2):158–165. https://doi.org/10.3945/an.111.001644
Sheng L, Zhu GL, Tong QY (2013) Mechanism study of Tween 80 enhancing the pullulan production by Aureobasidium pullulans. Carbohydr Polym 97(1):121–123. https://doi.org/10.1016/j.carbpol.2013.04.058
Singh A, Van Hamme JD, Ward OP (2007) Surfactants in microbiology and biotechnology: Part 2. Application aspects. Biotechnol Adv 25(1):99–121. https://doi.org/10.1016/j.biotechadv.2006.10.004
Tani Y, Asahi S, Yamada H (1986) Menaquinone (vitamin K2)-6 production by mutants of Flavobacterium meningosepticum. J Nutr Sci Vitaminol (tokyo) 32(2):137–145. https://doi.org/10.3177/jnsv.32.137
Tu GW, Wang YK, Ji YC, Zou X (2015) The effect of Tween 80 on the polymalic acid and pullulan production by Aureobasidium pullulans CCTCC M2012223. World J Microbiol Biotechnol 31(1):219–226. https://doi.org/10.1007/s11274-014-1779-9
Vos M, Esposito G, Edirisinghe JN, Vilain S, Haddad DM, Slabbaert JR, Van Meensel S, Schaap O, De Strooper B, Meganathan R, Morais VA, Verstreken P (2012) Vitamin K2 is a mitochondrial electron carrier that rescues pink1 deficiency. Science 336(6086):1306–1310. https://doi.org/10.1126/science.1218632
Wang H, Liu H, Wang L, Zhao GH, Tang HF, Sun XW, Ni WF, Yang Q, Wang P, Zheng ZM (2019) Improvement of menaquinone-7 production by Bacillus subtilis natto in a novel residue-free medium by increasing the redox potential. Appl Microbiol Biotechnol 103(18):7519–7535. https://doi.org/10.1007/s00253-019-10044-5
Wu J, Li W, Zhao SG, Qian SH, Wang Z, Zhou MJ, Hu WS, WangJ HuLX, LiuY X, ZL, (2021) Site-directed mutagenesis of the quorum-sensing transcriptional regulator SinR affects the biosynthesis of menaquinone in Bacillus subtilis. Microb Cell Fact 20(1):113
Zhang BB, Cheung PC (2011) A mechanistic study of the enhancing effect of Tween 80 on the mycelial growth and exopolysaccharide production by Pleurotus tuber-regium. Bioresour Technol 102(17):8323–8326. https://doi.org/10.1016/j.biortech.2011.06.021
Zhang ZB, Zeng GM, Shi JG, Liu J, Yang WC (2006) Effect of Tween 80 and rhamnolipid on the production of protease from Pseudomonas Aeruginosa and Bacillus Subtilis. Acta Sci Circum 26:1152–1158. https://doi.org/10.3321/j.issn:0253-2468.2006.07.018
Zhao CL, Wan YP, Tang GX, Jin Q, Zhang HL, Xu ZN (2020) Comparison of different fermentation processes for the vitamin K2 (Menaquinone-7) production by a novel Bacillus velezensis ND strain. Process Biochem 102(9):33–41. https://doi.org/10.1016/j.procbio.2020.11.029
Acknowledgements
The study was supported by the National Nature Science Foundation of China (Nos. 31871781, 31772081), Science and technology project of Wuhu (2020yf62), the National Undergraduate Innovation and Entrepreneurship Program (No. 201910363042) and Anhui Provincial Undergraduate Innovation and Entrepreneurship Program (No. S202010363255).
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by MZ, JW, LH, WH, JH, XH, XG, YL, ZX and YL. The first draft of the manuscript was written by MZ, JW and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Zhou, Mj., Jing Wu, Hu, Lx. et al. Enhanced vitamin K2 production by engineered Bacillus subtilis during leakage fermentation. World J Microbiol Biotechnol 39, 224 (2023). https://doi.org/10.1007/s11274-023-03671-8
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DOI: https://doi.org/10.1007/s11274-023-03671-8