Menaquinone-7 production in engineered Escherichia coli

Gao, Quanxiu; Chen, Hao; Wang, Wenzhao; Huang, Jianzhong; Tao, Yong; Lin, Bingcheng

doi:10.1007/s11274-020-02880-9

Cited by 16 publications

(11 citation statements)

References 19 publications

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“…Previous research has shown that E. coli mainly synthesises one type of vitamin K2, menaquinone-8 (MK-8), under micro-anaerobic conditions [ 29 ]. MK-7 can also be produced by heterogeneous heptaprenyl pyrophosphate synthetase (HepPPS) [ 37 ]. In this study, LC–MS revealed the accumulation of MK-6, -7, and -8 by strain J01 (Additional file 1 : Fig.…”

Section: Resultsmentioning

confidence: 99%

Engineering microbial consortia of Elizabethkingia meningoseptica and Escherichia coli strains for the biosynthesis of vitamin K2

et al. 2022

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Background The study and application of microbial consortia are topics of interest in the fields of metabolic engineering and synthetic biology. In this study, we report the design and optimisation of Elizabethkingia meningoseptica and Escherichia coli co-culture, which bypass certain limitations found during the molecular modification of E. meningoseptica, such as resistance to many antibiotics and fewer available molecular tools. Results The octaprenyl pyrophosphate synthase from E. meningoseptica sp. F2 (EmOPPS) was expressed, purified, and identified in the present study. Then, owing to the low vitamin K2 production by E. coli or E. meningoseptica sp. F2 monoculture, we introduced the E. meningoseptica and E. coli co-culture strategy to improve vitamin K2 biosynthesis. We achieved production titres of 32 mg/L by introducing vitamin K2 synthesis-related genes from E. meningoseptica sp. F2 into E. coli, which were approximately three-fold more than the titre achieved with E. meningoseptica sp. F2 monoculture. This study establishes a foundation for further engineering of MK-n (n = 4, 5, 6, 7, 8) in a co-cultivation system of E. meningoseptica and E. coli. Finally, we analysed the surface morphology, esterase activity, and membrane permeability of these microbial consortia using scanning electron microscopy, confocal laser scanning microscopy, and flow cytometry, respectively. The results showed that the co-cultured bacteria were closely linked and that lipase activity and membrane permeability improved, which may be conducive to the exchange of substances between bacteria. Conclusions Our results demonstrated that co-culture engineering can be a useful method in the broad field of metabolic engineering of strains with restricted molecular modifications.

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Section: Resultsmentioning

confidence: 99%

Engineering microbial consortia of Elizabethkingia meningoseptica and Escherichia coli strains for the biosynthesis of vitamin K2

et al. 2022

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“…A previous study showed that high titer of ubiquinone was produced in engineered MK-7 producing E. coli strains, and the knockout of the competitive pathways of MK-7 biosynthesis could not increase the titer of MK-7 . In the attempt to synthesize MK-7 under aerobic conditions, the flux of MK-7 biosynthesis needs to be enhanced without blocking the pathway of ubiquinone synthesis.…”

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confidence: 78%

“…coli strain which just proved the concept that MK-7 could be produced in E. coli . In this study, to achieve an E.…”

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confidence: 99%

Highly Efficient Production of Menaquinone-7 from Glucose by Metabolically Engineered Escherichia coli

et al. 2021

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Menaquinone-7 (MK-7) possesses wide health and medical value, and the market demand for MK-7 has increased. Metabolic engineering for MK-7 production in Escherichia coli still remains challenging due to the characteristics of the competing quinone synthesis, and cells mainly synthesized menaquinones under anaerobic conditions. To increase the production of MK-7 in engineered E. coli strains under aerobic conditions, we divided the whole MK-7 biosynthetic pathway into three modules (MVA pathway, DHNA pathway, and MK-7 pathway) and systematically optimized each of them. First, by screening and enhancing Idi expression, the amounts of MK-7/DMK-7 increased significantly. Then, in the MK-7 pathway, by combinatorial overexpression of endogenous MenA and exogenous UbiE, and fine-tuning the expression of HepPPS, MenA, and UbiE, 70 μM MK-7 was achieved. Third, the DHNA synthetic pathway was enhanced, and 157 μM MK-7 was achieved. By the combinational metabolic engineering strategies and membrane engineering, an efficient metabolic engineered E. coli strain for MK-7 synthesis was developed, and 200 μM (129 mg/L) MK-7 was obtained in shake flask experiment, representing a 306-fold increase compared to the starting strain. In the scale-up fermentation, 2074 μM (1350 mg/L) MK-7 was achieved after 52 h fermentation with a productivity of 26 mg/L/h. This is the highest titer of MK-7 ever reported. This study offers an alternative method for MK-7 production from biorenewable feedstock (glucose) by engineered E. coli. The high titer of our process should make it a promising cost-effective resource for MK-7.

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“…This study was carried out to produce engineered E. coli by introducing heptaprenyl pyrophosphatase synthetase (HepPPS) enzyme derived from B. subtilis . Further, production of vitamin K2-7 was enhanced by overexpressing HepPPS and optimizing the expression of enzymes in mevalonic acid pathway in order to eliminate synthesis of metabolites other than those involved in K2-7 pathway ( Gao et al, 2020 ). In another study Gao et al investigated an alternative procedure for highly efficient production of vitamin K2-7 by using metabolically engineered E. coli under aerobic conditions ( Gao et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Molecular Pathways and Roles for Vitamin K2-7 as a Health-Beneficial Nutraceutical: Challenges and Opportunities

Jadhav¹,

Ajgaonkar²,

Saha³

et al. 2022

Front. Pharmacol.

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Vitamin K2-7, also known as menaquinone-7 (MK-7) is a form of vitamin K that has health-beneficial effects in osteoporosis, cardiovascular disease, inflammation, cancer, Alzheimer’s disease, diabetes and peripheral neuropathy. Compared to vitamin K1 (phylloquinone), K2-7 is absorbed more readily and is more bioavailable. Clinical studies have unequivocally demonstrated the utility of vitamin K2-7 supplementation in ameliorating peripheral neuropathy, reducing bone fracture risk and improving cardiovascular health. We examine how undercarboxylated osteocalcin (ucOC) and matrix Gla protein (ucMGP) are converted to carboxylated forms (cOC and cMGP respectively) by K2-7 acting as a cofactor, thus facilitating the deposition of calcium in bones and preventing vascular calcification. K2-7 is beneficial in managing bone loss because it upregulates osteoprotegerin which is a decoy receptor for RANK ligand (RANKL) thus inhibiting bone resorption. We also review the evidence for the health-beneficial outcomes of K2-7 in diabetes, peripheral neuropathy and Alzheimer’s disease. In addition, we discuss the K2-7-mediated suppression of growth in cancer cells via cell-cycle arrest, autophagy and apoptosis. The mechanistic basis for the disease-modulating effects of K2-7 is mediated through various signal transduction pathways such as PI3K/AKT, MAP Kinase, JAK/STAT, NF-κB, etc. Interestingly, K2-7 is also responsible for suppression of proinflammatory mediators such as IL-1α, IL-1β and TNF-α. We elucidate various genes modulated by K2-7 as well as the clinical pharmacometrics of vitamin K2-7 including K2-7-mediated pharmacokinetics/pharmacodynamics (PK/PD). Further, we discuss the current status of clinical trials on K2-7 that shed light on dosing strategies for maximum health benefits. Taken together, this is a synthetic review that delineates the health-beneficial effects of K2-7 in a clinical setting, highlights the molecular basis for these effects, elucidates the clinical pharmacokinetics of K2-7, and underscores the need for K2-7 supplementation in the global diet.

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Menaquinone-7 production in engineered Escherichia coli

Cited by 16 publications

References 19 publications

Engineering microbial consortia of Elizabethkingia meningoseptica and Escherichia coli strains for the biosynthesis of vitamin K2

Engineering microbial consortia of Elizabethkingia meningoseptica and Escherichia coli strains for the biosynthesis of vitamin K2

Highly Efficient Production of Menaquinone-7 from Glucose by Metabolically Engineered Escherichia coli

Molecular Pathways and Roles for Vitamin K2-7 as a Health-Beneficial Nutraceutical: Challenges and Opportunities

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