Characterization of feedback-resistant mevalonate kinases from the methanogenic archaeons Methanosaeta concilii and Methanocella paludicola

Kazieva, Ekaterina; Yamamoto, Yoko; Tajima, Yoshinori; Yokoyama, Keiichi; Katashkina, J. I.; Nishio, Yoshihiko

doi:10.1099/mic.0.000510

Cited by 27 publications

(27 citation statements)

References 49 publications

(40 reference statements)

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“…Recently, feedback-resistant MvaK1 enzymes were identified and characterized from Methanosaeta concilii (McMvaK) and Methanocella paludicola (MpMvaK) [20]. The McMvaK and MpMvaK enzymes not only showed feedback resistance to DMAPP, IPP, GPP, and FPP, but also exhibited 4.9-and 5.5-fold higher affinity to MVA, respectively, than MmMvaK1 [20]. Therefore, these MvaK1 enzymes may enable the enhancement of (−)-α-bisabolol production in engineered E. coli.…”

Section: Discussionmentioning

confidence: 99%

Enhanced (−)-α-Bisabolol Productivity by Efficient Conversion of Mevalonate in Escherichia coli

Kim

Seong

et al. 2019

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(−)-α-Bisabolol, a naturally occurring sesquiterpene alcohol, has been used in pharmaceuticals and cosmetics owing to its beneficial effects on inflammation and skin healing. Previously, we reported the high production of (−)-α-bisabolol by fed-batch fermentation using engineered Escherichia coli (E. coli) expressing the exogenous mevalonate (MVA) pathway genes. The productivity of (−)-α-bisabolol must be improved before industrial application. Here, we report enhancement of initial (−)-α-bisabolol productivity to 3-fold higher than that observed in our previous study. We first harnessed a farnesyl pyrophosphate (FPP)-resistant mevalonate kinase 1 (MvaK1) from an archaeon Methanosarcina mazei (M. mazei) to create a more efficient heterologous MVA pathway that produces (−)-α-bisabolol in the engineered E. coli. The resulting strain produced 1.7-fold higher (−)-α-bisabolol relative to the strain expressing a feedback-inhibitory MvaK1 from Staphylococcus aureus (S. aureus). Next, to efficiently convert accumulated MVA to (−)-α-bisabolol, we additionally overexpressed genes involved in the lower MVA mevalonate pathway in E. coli containing the entire MVA pathway genes. (−)-α-Bisabolol production increased by 1.8-fold with reduction of MVA accumulation, relative to the control strain. Finally, we optimized the fermentation conditions including inducer concentration, aeration and enzymatic cofactor. The strain was able to produce 8.5 g/L of (−)-α-bisabolol with an initial productivity of 0.12 g/L h in the optimal fed-batch fermentation. Thus, the microbial production of (−)-α-bisabolol would be an economically viable bioprocess for its industrial application.

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Section: Discussionmentioning

confidence: 99%

Enhanced (−)-α-Bisabolol Productivity by Efficient Conversion of Mevalonate in Escherichia coli

Kim

Seong

et al. 2019

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“…Directed evolution (Anthony et al., 2009) and overexpression (Chen et al., 2018b) of MK has been applied to increase yields of IPP downstream products. In addition, several feedback-resistant MKs have been identified and characterized (Kazieva et al., 2017; Primak et al., 2011), which can be expressed in other microorganisms for carotenoid overproduction. Similarly, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR1) can be feedback inhibited by FPP, which limits the carotenoid yield.…”

Section: Modular Engineering For Carotenoids Productionmentioning

confidence: 99%

Modular engineering for microbial production of carotenoids

Swofford

Sinskey

2020

Metabolic Engineering Communications

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There is an increasing demand for carotenoids due to their applications in the food, flavor, pharmaceutical and feed industries, however, the extraction and synthesis of these compounds can be expensive and technically challenging. Microbial production of carotenoids provides an attractive alternative to the negative environmental impacts and cost of chemical synthesis or direct extraction from plants. Metabolic engineering and synthetic biology approaches have been widely utilized to reconstruct and optimize pathways for carotenoid overproduction in microorganisms. This review summarizes the current advances in microbial engineering for carotenoid production and divides the carotenoid biosynthesis building blocks into four distinct metabolic modules: 1) central carbon metabolism, 2) cofactor metabolism, 3) isoprene supplement metabolism and 4) carotenoid biosynthesis. These four modules focus on redirecting carbon flux and optimizing cofactor supplements for isoprene precursors needed for carotenoid synthesis. Future perspectives are also discussed to provide insights into microbial engineering principles for overproduction of carotenoids.

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“…A codon‐optimized DNA fragment of the mvk gene encoding Methanocella paludicola strain SANAE MVA kinase (MVK; GenBank no. ) (Kazieva et al ) was chemically synthesized and cloned into pUC57 to yield pUC57‐ mvk (GenScript). The fragment containing the mvk gene was PCR‐amplified from pUC57‐ mvk using primers mvk‐Fw/mvk‐Rv and digested with Pst I/ Kpn I, after which this fragment was ligated with Pst I/ Kpn I‐digested pAH162‐P tac to yield pAH162‐P tac ‐ mvk .…”

Section: Methodsmentioning

confidence: 99%

“…MS experiments were conducted in the selected ion‐monitoring mode to detect ion m/z 147. The chemical standard solutions of MVA (10, 40 and 80 mg l −1 ) were prepared from mevalonolactone (ADEKA, Tokyo, Japan) according to a previously reported protocol (Kazieva et al ). Peak areas were converted to MVA concentration with a calibration curve.…”

Section: Methodsmentioning

confidence: 99%

Application of inorganic phosphate limitation to efficient isoprene production in Pantoea ananatis

et al. 2019

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Aims Establishment of an efficient isoprene fermentation process by adopting inorganic phosphate limitation as the trigger to direct metabolic flux to the isoprene synthetic pathway. Methods and Results We constructed isoprene‐producing strains of Pantoea ananatis (a member of the Enterobacteriaceae family) by integrating a heterologous mevalonate pathway and a metabolic switch that senses external inorganic phosphate (Pi) levels. This metabolic switch enabled dual‐phase isoprene production, where the initial cell growth phase under Pi‐saturating conditions was uncoupled from the subsequent isoprene production phase under Pi‐limiting conditions. In fed‐batch fermentation using our best strain (SWITCH‐PphoC/pIspSM) in a 1‐l bioreactor, isoprene concentration in the off‐gas was maintained between 300 and 460 ppm during the production phase and at 20 ppm during the cell growth phase, respectively. The strain SWITCH‐PphoC/pIspSM produced totally 2·5 g l−1 of isoprene from glucose with a 1·8% volumetric yield in 48 h. Conclusions This proof‐of‐concept study demonstrated that our Pi‐dependent dual‐phase production system using a P. ananatis strain as a producer has potential for industrial‐scale isoprene fermentation. Significance and Impact of the Study This Pi‐dependent dual‐phase fermentation process could be an attractive and economically viable option for the production of various commercially valuable isoprenoids.

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Characterization of feedback-resistant mevalonate kinases from the methanogenic archaeons Methanosaeta concilii and Methanocella paludicola

Cited by 27 publications

References 49 publications

Enhanced (−)-α-Bisabolol Productivity by Efficient Conversion of Mevalonate in Escherichia coli

Enhanced (−)-α-Bisabolol Productivity by Efficient Conversion of Mevalonate in Escherichia coli

Modular engineering for microbial production of carotenoids

Application of inorganic phosphate limitation to efficient isoprene production in Pantoea ananatis

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